GERMINATION OF HIBISCUS SABDARIFFA L SEEDS AFTER STORAGE UNDER DIFFERENT ENVIRONMENTS
Jovinia Binti Jowinis
QK 661 Bachelor of Science with Honours J86 (Plant Resource Science and Management) 2005 2005
Pusat Khidmat Makfumat Akadeu UNIVERSITI MALAYSIA SARAWA
94100 KOla Samarahan
GERMINATION OF HIBISCUS SABDARIFF A L SEEDS AFTER STORAGE UNDER DIFFERENT ENVIRONMENTS
PKHIDMA TMAKLUMATAKADEMIK UIlIMAS
IIIII 1111 In11111111111 1000128296
JOVINIA BINTI JOWINIS
This report is submitted in partial fulfillment of the requirements for the degree of Bachelor of Science with Honours
(Plant Resource Science and Management)
PLANT RESOURCE SCIENCE AND MANAGEMENT FACULTY OF RESOURCE SCIENCE AND TECHNOLOGY
UNIVERSITI MALAYSIA SARAWAK
2005
ACKNOWLEDGEMENTS
I would like to express my gratitude and appreciation to my project supervisor Dr Petrus
Bulan Lecturer Faculty of Resource Science and Technology University Malaysia Sarawak
Kota Samarahan for his invaluable guidance encouragement and dedication in the execution
and completion of this study and for his patient and assistance during the preparation of this
manuscript Thanks to Dr Ismail Jusoh for his guidance in SPSS analysis
My appreciation to Mr Amin ak Manggi and Mr Azis bin Ajim of the Cryopreservation
Laboratory Faculty of Resource Science and Technology Universiti Malaysia Sarawak for
their assistance My sincere gratitude to all my friends and colleagues especially Laurina
Jacklyn Teo Shirley Maurice Labanjun Ida Rani Ali and Siti Khuzaimah Ahmad for their
understanding and helping me in one way or another My deepest appreciation and love to my
parents Mr Jowinis Lawadi and Mrs Margaret Wong for their encouragement in completion
of this study I am very grateful to the Gulf amp Pacific Industries Sdn Bhd Company for
supplying the seed material for this study
11
TABLE OF CONTENTS
Contents Page
Acknowledgement 11
List of Tables and Figures v
Abstract viii
Abstrak viii
CHAPTER 1 INTRODUCTION
11 Background 1
12 Medicinal Uses 2
13 Food Uses 4
14 Other Uses 5
15 Problem Statement 6
16 Objectives 6
CHAPTER 2 LITERATURE REVIEW 7
21 Seed Gennination 7
22 Seed Storage 9
23 Seed Deterioration 11
CHAPTER 3 MATERIAL AND METHOD 14
31 Material 14
32 Method 14
321 Moisture Content Test 14
322 Gennination Test 15
323 Seed Storage 16
324 Data Analysis 16
111
I
CHAPTER 4 RESULT AND DISCUSSION 17
41 Initial Seed quality 17
42 Seed Moisture Content and Germination 17
43 Seed Storage 19
431 Seed Moisture Content 19
432 Seed Germination 21
44 Regression Analysis 27
CHAPTER 5 CONCLUSION AND RECOMMENDATION 32
CHAPTER 6 REFERENCES 34
CllAPTER 7 APPENDIX 38
IV
LIST OF TABLES AND FIGURES
1 Effect of storage temperature on moisture content and
germination ofH sabdariffa seeds 24
2 Means of moisture content and germination ofH sabdariffa seeds in four different treatments 26
3 Moisture Content Test of H sabdariffa seeds 38
4 Germination Test ofH sabdariffa seeds 38
5 Moisture content and germination ofH sabdariffa seeds 38
6 Moisture Content ofH sabdariffa seeds after 14 Days of Storage 39
7 ANOVA analysis for seeds moisture content 39
8 Germination ofH sabdariffa seeds after 14 days of storage 40
9 ANOVA analysis for H sabdariffa seeds germination 40
10 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in ambient room 41
11 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in ambient room 41
12 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in ambient room 41
13 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in air-conditioned room 42
14 Regression Analysis (ANOVA)for germination of H sabdariffa seeds stored in air-conditioned room 42
15 Regression Analysis (Coefficient) for germination of H sabdariffa seeds stored in air-conditioned room 42
16 Regression Analysis (Model Summary) for germination of sabdariffa seeds stored in refrigerator 43
v
17 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in refrigerator 43
18 Regression Analysis (Coefficient) for germination ofH sabdariffa seed stored in refrigerator 43
19 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in incubator 44
20 Regression Analysis (ANOV A) for germination ofH sabdariffa seeds stored in incubator 44
21 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in incubator 44
22 ANOV A analysis between factors and period of storage 45
23 ANOVA analysis between factors and treatment 45
VI
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
Pusat Khidmat Makfumat Akadeu UNIVERSITI MALAYSIA SARAWA
94100 KOla Samarahan
GERMINATION OF HIBISCUS SABDARIFF A L SEEDS AFTER STORAGE UNDER DIFFERENT ENVIRONMENTS
PKHIDMA TMAKLUMATAKADEMIK UIlIMAS
IIIII 1111 In11111111111 1000128296
JOVINIA BINTI JOWINIS
This report is submitted in partial fulfillment of the requirements for the degree of Bachelor of Science with Honours
(Plant Resource Science and Management)
PLANT RESOURCE SCIENCE AND MANAGEMENT FACULTY OF RESOURCE SCIENCE AND TECHNOLOGY
UNIVERSITI MALAYSIA SARAWAK
2005
ACKNOWLEDGEMENTS
I would like to express my gratitude and appreciation to my project supervisor Dr Petrus
Bulan Lecturer Faculty of Resource Science and Technology University Malaysia Sarawak
Kota Samarahan for his invaluable guidance encouragement and dedication in the execution
and completion of this study and for his patient and assistance during the preparation of this
manuscript Thanks to Dr Ismail Jusoh for his guidance in SPSS analysis
My appreciation to Mr Amin ak Manggi and Mr Azis bin Ajim of the Cryopreservation
Laboratory Faculty of Resource Science and Technology Universiti Malaysia Sarawak for
their assistance My sincere gratitude to all my friends and colleagues especially Laurina
Jacklyn Teo Shirley Maurice Labanjun Ida Rani Ali and Siti Khuzaimah Ahmad for their
understanding and helping me in one way or another My deepest appreciation and love to my
parents Mr Jowinis Lawadi and Mrs Margaret Wong for their encouragement in completion
of this study I am very grateful to the Gulf amp Pacific Industries Sdn Bhd Company for
supplying the seed material for this study
11
TABLE OF CONTENTS
Contents Page
Acknowledgement 11
List of Tables and Figures v
Abstract viii
Abstrak viii
CHAPTER 1 INTRODUCTION
11 Background 1
12 Medicinal Uses 2
13 Food Uses 4
14 Other Uses 5
15 Problem Statement 6
16 Objectives 6
CHAPTER 2 LITERATURE REVIEW 7
21 Seed Gennination 7
22 Seed Storage 9
23 Seed Deterioration 11
CHAPTER 3 MATERIAL AND METHOD 14
31 Material 14
32 Method 14
321 Moisture Content Test 14
322 Gennination Test 15
323 Seed Storage 16
324 Data Analysis 16
111
I
CHAPTER 4 RESULT AND DISCUSSION 17
41 Initial Seed quality 17
42 Seed Moisture Content and Germination 17
43 Seed Storage 19
431 Seed Moisture Content 19
432 Seed Germination 21
44 Regression Analysis 27
CHAPTER 5 CONCLUSION AND RECOMMENDATION 32
CHAPTER 6 REFERENCES 34
CllAPTER 7 APPENDIX 38
IV
LIST OF TABLES AND FIGURES
1 Effect of storage temperature on moisture content and
germination ofH sabdariffa seeds 24
2 Means of moisture content and germination ofH sabdariffa seeds in four different treatments 26
3 Moisture Content Test of H sabdariffa seeds 38
4 Germination Test ofH sabdariffa seeds 38
5 Moisture content and germination ofH sabdariffa seeds 38
6 Moisture Content ofH sabdariffa seeds after 14 Days of Storage 39
7 ANOVA analysis for seeds moisture content 39
8 Germination ofH sabdariffa seeds after 14 days of storage 40
9 ANOVA analysis for H sabdariffa seeds germination 40
10 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in ambient room 41
11 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in ambient room 41
12 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in ambient room 41
13 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in air-conditioned room 42
14 Regression Analysis (ANOVA)for germination of H sabdariffa seeds stored in air-conditioned room 42
15 Regression Analysis (Coefficient) for germination of H sabdariffa seeds stored in air-conditioned room 42
16 Regression Analysis (Model Summary) for germination of sabdariffa seeds stored in refrigerator 43
v
17 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in refrigerator 43
18 Regression Analysis (Coefficient) for germination ofH sabdariffa seed stored in refrigerator 43
19 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in incubator 44
20 Regression Analysis (ANOV A) for germination ofH sabdariffa seeds stored in incubator 44
21 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in incubator 44
22 ANOV A analysis between factors and period of storage 45
23 ANOVA analysis between factors and treatment 45
VI
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
ACKNOWLEDGEMENTS
I would like to express my gratitude and appreciation to my project supervisor Dr Petrus
Bulan Lecturer Faculty of Resource Science and Technology University Malaysia Sarawak
Kota Samarahan for his invaluable guidance encouragement and dedication in the execution
and completion of this study and for his patient and assistance during the preparation of this
manuscript Thanks to Dr Ismail Jusoh for his guidance in SPSS analysis
My appreciation to Mr Amin ak Manggi and Mr Azis bin Ajim of the Cryopreservation
Laboratory Faculty of Resource Science and Technology Universiti Malaysia Sarawak for
their assistance My sincere gratitude to all my friends and colleagues especially Laurina
Jacklyn Teo Shirley Maurice Labanjun Ida Rani Ali and Siti Khuzaimah Ahmad for their
understanding and helping me in one way or another My deepest appreciation and love to my
parents Mr Jowinis Lawadi and Mrs Margaret Wong for their encouragement in completion
of this study I am very grateful to the Gulf amp Pacific Industries Sdn Bhd Company for
supplying the seed material for this study
11
TABLE OF CONTENTS
Contents Page
Acknowledgement 11
List of Tables and Figures v
Abstract viii
Abstrak viii
CHAPTER 1 INTRODUCTION
11 Background 1
12 Medicinal Uses 2
13 Food Uses 4
14 Other Uses 5
15 Problem Statement 6
16 Objectives 6
CHAPTER 2 LITERATURE REVIEW 7
21 Seed Gennination 7
22 Seed Storage 9
23 Seed Deterioration 11
CHAPTER 3 MATERIAL AND METHOD 14
31 Material 14
32 Method 14
321 Moisture Content Test 14
322 Gennination Test 15
323 Seed Storage 16
324 Data Analysis 16
111
I
CHAPTER 4 RESULT AND DISCUSSION 17
41 Initial Seed quality 17
42 Seed Moisture Content and Germination 17
43 Seed Storage 19
431 Seed Moisture Content 19
432 Seed Germination 21
44 Regression Analysis 27
CHAPTER 5 CONCLUSION AND RECOMMENDATION 32
CHAPTER 6 REFERENCES 34
CllAPTER 7 APPENDIX 38
IV
LIST OF TABLES AND FIGURES
1 Effect of storage temperature on moisture content and
germination ofH sabdariffa seeds 24
2 Means of moisture content and germination ofH sabdariffa seeds in four different treatments 26
3 Moisture Content Test of H sabdariffa seeds 38
4 Germination Test ofH sabdariffa seeds 38
5 Moisture content and germination ofH sabdariffa seeds 38
6 Moisture Content ofH sabdariffa seeds after 14 Days of Storage 39
7 ANOVA analysis for seeds moisture content 39
8 Germination ofH sabdariffa seeds after 14 days of storage 40
9 ANOVA analysis for H sabdariffa seeds germination 40
10 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in ambient room 41
11 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in ambient room 41
12 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in ambient room 41
13 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in air-conditioned room 42
14 Regression Analysis (ANOVA)for germination of H sabdariffa seeds stored in air-conditioned room 42
15 Regression Analysis (Coefficient) for germination of H sabdariffa seeds stored in air-conditioned room 42
16 Regression Analysis (Model Summary) for germination of sabdariffa seeds stored in refrigerator 43
v
17 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in refrigerator 43
18 Regression Analysis (Coefficient) for germination ofH sabdariffa seed stored in refrigerator 43
19 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in incubator 44
20 Regression Analysis (ANOV A) for germination ofH sabdariffa seeds stored in incubator 44
21 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in incubator 44
22 ANOV A analysis between factors and period of storage 45
23 ANOVA analysis between factors and treatment 45
VI
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
TABLE OF CONTENTS
Contents Page
Acknowledgement 11
List of Tables and Figures v
Abstract viii
Abstrak viii
CHAPTER 1 INTRODUCTION
11 Background 1
12 Medicinal Uses 2
13 Food Uses 4
14 Other Uses 5
15 Problem Statement 6
16 Objectives 6
CHAPTER 2 LITERATURE REVIEW 7
21 Seed Gennination 7
22 Seed Storage 9
23 Seed Deterioration 11
CHAPTER 3 MATERIAL AND METHOD 14
31 Material 14
32 Method 14
321 Moisture Content Test 14
322 Gennination Test 15
323 Seed Storage 16
324 Data Analysis 16
111
I
CHAPTER 4 RESULT AND DISCUSSION 17
41 Initial Seed quality 17
42 Seed Moisture Content and Germination 17
43 Seed Storage 19
431 Seed Moisture Content 19
432 Seed Germination 21
44 Regression Analysis 27
CHAPTER 5 CONCLUSION AND RECOMMENDATION 32
CHAPTER 6 REFERENCES 34
CllAPTER 7 APPENDIX 38
IV
LIST OF TABLES AND FIGURES
1 Effect of storage temperature on moisture content and
germination ofH sabdariffa seeds 24
2 Means of moisture content and germination ofH sabdariffa seeds in four different treatments 26
3 Moisture Content Test of H sabdariffa seeds 38
4 Germination Test ofH sabdariffa seeds 38
5 Moisture content and germination ofH sabdariffa seeds 38
6 Moisture Content ofH sabdariffa seeds after 14 Days of Storage 39
7 ANOVA analysis for seeds moisture content 39
8 Germination ofH sabdariffa seeds after 14 days of storage 40
9 ANOVA analysis for H sabdariffa seeds germination 40
10 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in ambient room 41
11 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in ambient room 41
12 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in ambient room 41
13 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in air-conditioned room 42
14 Regression Analysis (ANOVA)for germination of H sabdariffa seeds stored in air-conditioned room 42
15 Regression Analysis (Coefficient) for germination of H sabdariffa seeds stored in air-conditioned room 42
16 Regression Analysis (Model Summary) for germination of sabdariffa seeds stored in refrigerator 43
v
17 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in refrigerator 43
18 Regression Analysis (Coefficient) for germination ofH sabdariffa seed stored in refrigerator 43
19 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in incubator 44
20 Regression Analysis (ANOV A) for germination ofH sabdariffa seeds stored in incubator 44
21 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in incubator 44
22 ANOV A analysis between factors and period of storage 45
23 ANOVA analysis between factors and treatment 45
VI
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
I
CHAPTER 4 RESULT AND DISCUSSION 17
41 Initial Seed quality 17
42 Seed Moisture Content and Germination 17
43 Seed Storage 19
431 Seed Moisture Content 19
432 Seed Germination 21
44 Regression Analysis 27
CHAPTER 5 CONCLUSION AND RECOMMENDATION 32
CHAPTER 6 REFERENCES 34
CllAPTER 7 APPENDIX 38
IV
LIST OF TABLES AND FIGURES
1 Effect of storage temperature on moisture content and
germination ofH sabdariffa seeds 24
2 Means of moisture content and germination ofH sabdariffa seeds in four different treatments 26
3 Moisture Content Test of H sabdariffa seeds 38
4 Germination Test ofH sabdariffa seeds 38
5 Moisture content and germination ofH sabdariffa seeds 38
6 Moisture Content ofH sabdariffa seeds after 14 Days of Storage 39
7 ANOVA analysis for seeds moisture content 39
8 Germination ofH sabdariffa seeds after 14 days of storage 40
9 ANOVA analysis for H sabdariffa seeds germination 40
10 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in ambient room 41
11 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in ambient room 41
12 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in ambient room 41
13 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in air-conditioned room 42
14 Regression Analysis (ANOVA)for germination of H sabdariffa seeds stored in air-conditioned room 42
15 Regression Analysis (Coefficient) for germination of H sabdariffa seeds stored in air-conditioned room 42
16 Regression Analysis (Model Summary) for germination of sabdariffa seeds stored in refrigerator 43
v
17 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in refrigerator 43
18 Regression Analysis (Coefficient) for germination ofH sabdariffa seed stored in refrigerator 43
19 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in incubator 44
20 Regression Analysis (ANOV A) for germination ofH sabdariffa seeds stored in incubator 44
21 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in incubator 44
22 ANOV A analysis between factors and period of storage 45
23 ANOVA analysis between factors and treatment 45
VI
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
LIST OF TABLES AND FIGURES
1 Effect of storage temperature on moisture content and
germination ofH sabdariffa seeds 24
2 Means of moisture content and germination ofH sabdariffa seeds in four different treatments 26
3 Moisture Content Test of H sabdariffa seeds 38
4 Germination Test ofH sabdariffa seeds 38
5 Moisture content and germination ofH sabdariffa seeds 38
6 Moisture Content ofH sabdariffa seeds after 14 Days of Storage 39
7 ANOVA analysis for seeds moisture content 39
8 Germination ofH sabdariffa seeds after 14 days of storage 40
9 ANOVA analysis for H sabdariffa seeds germination 40
10 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in ambient room 41
11 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in ambient room 41
12 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in ambient room 41
13 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in air-conditioned room 42
14 Regression Analysis (ANOVA)for germination of H sabdariffa seeds stored in air-conditioned room 42
15 Regression Analysis (Coefficient) for germination of H sabdariffa seeds stored in air-conditioned room 42
16 Regression Analysis (Model Summary) for germination of sabdariffa seeds stored in refrigerator 43
v
17 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in refrigerator 43
18 Regression Analysis (Coefficient) for germination ofH sabdariffa seed stored in refrigerator 43
19 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in incubator 44
20 Regression Analysis (ANOV A) for germination ofH sabdariffa seeds stored in incubator 44
21 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in incubator 44
22 ANOV A analysis between factors and period of storage 45
23 ANOVA analysis between factors and treatment 45
VI
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
17 Regression Analysis (ANOVA) for germination ofH sabdariffa seeds stored in refrigerator 43
18 Regression Analysis (Coefficient) for germination ofH sabdariffa seed stored in refrigerator 43
19 Regression Analysis (Model Summary) for germination of H sabdariffa seeds stored in incubator 44
20 Regression Analysis (ANOV A) for germination ofH sabdariffa seeds stored in incubator 44
21 Regression Analysis (Coefficient) for germination ofH sabdariffa seeds stored in incubator 44
22 ANOV A analysis between factors and period of storage 45
23 ANOVA analysis between factors and treatment 45
VI
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
Figure Page
1 Moisture content and gennination ofHibiscus sabdariffa seeds 17
2 Moisture content ofHibiscus sabdariffa seeds after stored in
four different environments for two weeks 19
3 Germination ofHibiscus sabdariffa seeds after stored in four
different environments for two weeks 21
4 Regression analysis for H sabdariffa seed stored in ambient
room (28-30degC) 27
5 Regression analysis for H sabdariffa seed stored in air-
conditioned room (22-23degC) 28
6 Regression analysis for H sabdariffa seed stored in refrigerator
(3-4degC) 29
7 Regression analysis for H sabdariffa seed stored in incubator
(38-40oC) 30
VB
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
Germination of Hibiscus sabdarifla after Storage under Different Environments
Jovinia Binti Jowinis
Plant Resource Science and Management Programme
Faculty ofResource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
A study was conducted to evaluate the effects of storage environments on germination of Hibiscus sabdariffa L seeds Seeds
with 89 of moisture content and 93 of germination were placed in four different environments ambient room (28-30degC)
air-conditioned room (22-23degC) refrigerator (3-4degC) and incubator (38-40degC) for 14 days Seed stored in air-conditioned
room gave the highest moisture content and germination with 770 and 91 respectively Storage in incubator showed
gradual decrement of moisture content and germination throughout the period of storage Generally moisture content and
germination percentages decreased with increase in storage period Regression analysis indicated that seeds stored in airshy
conditioned room can maintain its germinability up to 8 months and II days Results showed that storage in air-conditioned
room was a suitable condition to store H sabdariffa seeds
Keyword Hibiscus sabdariffa L critical moisture content storage
ABSTRAK
Satu kajian teah dijalanlum untuk meniai kesan persekitaran penyimpanan terhadap biji benih Hibiscus sabdariffa L Biji
benih dengan kandungan kelembapan 8900 dan percambahan 93 disimpan di dalam empat persekitaran yang berbeza
suhu bilik (28-3dC) bilik hawa dingin (22-23degC) peti sejuk (3-5degC) dan inkubator (38-4(fC) dalam jangleamasa 14 hari
Penyimpanan biji benih di daam bilik hawa dingin memberilean peratus leandungan kelembapan dan percambahan yang
paling tinggi iaitu 770 dan 91 Biji benih yang disimpan di dalam inkubator menunjukkan penurunan leandungan
kelembapan dan percambahan di sepanjang tempoh penyimpanan Secara umum peratus leandungan kelembapan dan
percambahan menurun dengan peningleatan tempoh penyimpanan Analisis regresi menunjulean penyimpanan biji benih di
dalam bilik hawa dingin boeh dilanjutlean sehingga 8 bulan 11 hari Penyimpanan biji benih di daam persekitaran bilik
hawa dingin didapati keadaan yang paling sesuai bagi biji benih H sabdariffa
Kola Hibiscus sabdariffa L leandungan keembapan kritilea penyimpanan
viii
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
CHAPTER ONE
INTRODUCTION
11 Background
Hibiscus sabdariffa L or known as Roselle belongs to family Malvaceae It is a short-day
annual plant that closely resembles cranberry (Vaccinium spp) in flavor (Morton 1987)
Roselle is known as asam susur in Malaysia Gamet Walanda and kasturi roriha in Indonesia
krachiap-daeng krachiap-prieo and phakkengkheng in Thailand and kubab and talingisag in
Philipines (Dasuki 2001) Vernacular names for H sabdariffa are sorrel rozelle red sorrel
Jamaica sorrel Indian sorrel Guinea sorrel sour-sour Queensland jelly plant jelly okra
lemon bush and Florida cranberry are H sabdariffas vernacular names ( Morton 1997) It is
a native of Africa but some do believed that it originated from India (Dasuki 2001)
Nowadays H sabdariffa is widely distributed in the tropics and subtropics of both
hemispheres and in many areas of West Indies and Central America (Morton 1987)
H sabdariffa is an erect annual herb 05-3 m tall stems glabrous purplish leaves broadly
ovate-orbicular variable 6-15 cm long lobes 3-5 oblong to lanceolate glabrous or
pubescent purplish petiole 5-10 cm long bracteoles of epicalyx 8-12 calyx after anthesis
becoming thick-fleshy 25-55 cm long distinctly longer than the fruit red corolla 3-5 cm
long not widely open pinkish to yellow with purple centre seed renifonn almost glabrous
black brown (Dasuki 2001)
The flower buds are actually seedpods enclosed in their fleshy calyxes The red pods or
ts and calyxes are fused and difficult to distinguish from one another (Apell 2003)
1
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
Roselle grows well on sandy loam and is tolerant to draught H sabdariffa is propagated
solely by seed Cultivation of roselle using cuttings produce short plants and yield of calyxes
is relatively low (Morton 1987)
Swollened and matured calyxes are harvested 75-115 days after planting When the crop
matured leaves and shoots are removed The removal of the shoots promotes branching
(Hallack nd) H sabdariffa is attacked mainly by root nematodes (Morton 1987) and
Phytophtora parasitica mainly in early stage of its development in the field (Follin 1996)
Alegbejo et al (2003) stated that H sabdariffa has a lot of potential as an industrial crop
Smallholder and resource poor farmers grow it It is popularly grown for its fiber content and
for its leaves calyxes roots and seeds which are used either for industrial medical
commercial or domestic uses
H sabdariffa and its parts can be utilized for many purposes (Duke 1983) It is used as health
products yielding fiber beverage edible foliage and an oil seed H sabdariffa is attracting
the attention of food and beverage manufacturers and pharmaceutical companies who feel it
may have possibilities as natural food product and as a color to replace some synthetic dyes
(Morton 1987)
12 Medicinal Uses
H sabdariffa is among the plant resources that are rich in calcium niacin riboflavin and iron
Morton (1987) agreed that in India Africa and Mexico all the above ground parts of the H
iffa plant are valued in native medicine The local infuse the leaves or calyxes to treat
diuretic choleretic febrifugal and hypotensive decreasing the viscosity of blood and
2
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
~ ~~-----------------------------------------------------------------------
stimulating intestinal peristalsis Some persons used it to relieve colds and heated leaves to
heal scars and wounds Heated leaves also applied to cracks in the feet and on boils and ulcers
to speed maturation A lotion made from H sabdariffa leaves is used on sores and wounds
According to a study by Duke (1983) H sabdariffa was reported to be antiseptic
aphrodisiac astringent cholagogue demulcent digestive diuretic emollient purgative
1efrigerant resolvent sedative stomachic and tonic It is also a folk remedy for abscesses
bilious conditions cancer cough debility dyspepsia dysuria fever hangover heart ailments
hypertension neurosis scurvy and strangury The drink made by placing the calyx in water
18 said to be a folk remedy for cancer Perry (1980) cites one study showing H sabdariffa
usefulness in ateriosc1erosis and as an intestinal antiseptic
Utilization of this plant is varied among countries In Burma the seed are used for debility and
the leaves as emollient Taiwanese regard the seed as diuretic laxative and tonic Philippines
jose the bitter root as an aperitive and tonic (Perry 1980) Angolans use the mucilaginous
leaves as an emollient and as a soothing cough remedy Central Africans poultice the leaves
on abscesses Other than that simulated ingestion of the H sabdariffa extract decreased the
ate of absorption of alcohol lessening the intensity of alcohol effects in chickens (Watt and
~reyer-Brandwijk) In Guinea they used the leaves as diuretic refringent and sedatives while
e fruits are antiascorbic leaves seeds and ripe calyces as diuretic and antiscorbutic calyces
Joiled in water used to treat billous attacks flowers are believed to contain gossypectin
anthocyanin and glucoside hibiscin which can decrease the viscosity of the blood and
tucing blood pressure (James 1983)
3
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
lI(Cgtsellle contains high antosianin and acid The nutritious calyxes contain high vitamin B 1
C and D H sabdariffa fiber remains in the intestine for beneficial fermentation in the
_u even though other nutrients from the food have been absorbed It helps the digestive
lIhsrem to function smoothly Fiber also helps to reduce cholesterol in the human body (Rhafa
This means H sabdariffa can help to maintain human body from cardiovascular diseases
which cholesterol plays a major role It is done by reducing fats in the body suitable for
_ under diet programme and weight reduction Tee et al (2002) agreed that roselle
nglltplay a role in the prevention of atherosclerosis and obesity
~I than has high potential in medicinal aspect this species also encouraging in food
PIi~J According to Watt and Breyer-Brandwijk (1962) H sabdariffa fruits are edible
calyxes can be consumed immediately by added into fruit salads Calyxes are stewed
sauce or juice syrup marmalade relish and chutney (Morton 1987) Calyxes are also
_OlU~ for tea and used to make jams and jellies (Hallack nd) Other than that it is used
to produce roselle wine refreshing beverages pudding and cakes meanwhile dry
used to produced sherbets butter pies tarts and other desserts (Duke 1983) H
OGfanlua syrup or sauce may be added to puddings cake frosting gelatins and salad
lIiltU11lI It can also be poured over gingerbread pancakes waffles or ice cream
species possesses 319 pectin and has been recommended as a source of pectin for
industry (Morton 1987) The efficacy of roselle as a functional food has been
_ lied lately especially for its antioxidant bioactivity (Tee et al 2002) Roselle leaves are
4
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
lDmetnnes eaten raw but are mostly cooked with Egusi (Cucmumeropsis edulis) to make a
~_ant sour tasting source usually served with fufu
(1987) agreed that the seeds are somewhat bitter but have been ground to a meal for
unlll food in Africa and also have been roasted as a substitute for coffee Duke (1983) stated
H sabdariffa seeds have been used as an aphrodisiac coffee substitute This means
the same as Eurycoma longifolia (Tongkat ali) extract function In the Caribbean the
fruits are brewed in water to make a refreshing cranberry-colored tea
mtiPnan dried H sabdariffa is added to commercial tea to flavor and color (Apell2003)
residue remaining after extraction of oil by parching soaking in water containing ashes
3 or 4 days and then pounding the seeds or by crushing and boiling them is eaten in soup
blended with bean meal in patties It is high in protein (Morton 1987)
i~lIttllo to Adamson et al (1975) H sabdarifJa is the only new crop introduced in the
MIlleill United States as a pulp source that shows a high level of resistance to nematodes H
1Mitn is cultivated primarily for the bast fiber obtained from the stems The fiber strands
to 15 m long are used for cordage and as a substitute for jute in the manufacture of burlap
non dehiscent and fleshy fruit containing anthocyanic pigments is largely used as tea and
beverage but also natural dye (Gassama-Dia et al 2004) Morton (1987)
_oned that the seeds are considered excellent feed for chickens The residue after oil
IIlIIIOn is valued as cattle feed when available in quantity
5
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
Malaysia H sabdarifJa is becoming more important as it has a wide range of benefits
_IV1l1TV Putra Malaysia (UPM) had commercialized H sabdarifJa in their plantation and
_nrvUPM import raw material of roselle from Sudan Africa to meet the required quota
~wt 2004) There are few H sabdarifJa plantations in Malaysia The resource in this
~ is insufficient to support production of H sabdarifJa The millers still import raw
tenal from overseas
maintenance and preservation of superior quality seeds are important for improved crop
rocI1llCtllon where 25 of the increase in agricultural production depends on seed quality
1981) The short age of raw material can be overcome by the use of quality seeds
in suitable storage environments Percentage of germination helped to exhibit quality of
seeds and a good indication of the potential field performance Seeds kept in storage of
environments will have poor quality and viability of the seeds
main objectives of this research were to analyze the influence of storage environments on
omlinlilticn of H sabdarifJa seeds and secondly to determine the suitable environmental
for storage of H sabdarifJa seeds for a long-term conservation
6
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
CHAPTER TWO
LITERATURE REVIEW
germination is a complex process involving many individual reactions and phases each
which is affected by temperature Because of its role in stand establishment seed
1m1D81ton remains a key to a modem agriculture (Copeland and McDonald 1995)
researchers further explained that germination is the activation of metabolic activity of
embryo leading to the emergence of a new seedling plant There are four important factors
lecting seed germination sufficient water adequate light suitable level of oxygen and
WUllWllC temperature
methods are commonly used to evaluate the quality of seeds after each treatment and
are moisture content and germination tests Germination test is conducted in order to
an idea ofhow the seeds will perform in the field
amppelland and McDonald (1995) agreed that germination test is most commonly used to
lllermirle seed viability It is universally accepted that seed germination and viability are
1liD1Ien~ as one They also stated that seed viability is highest at time of physiological
After physiological maturity the viability gradually declined and their longevity
_IlCleltl on the environmental to which they were exposed The rules of germination testing
only environmental conditions and dormancy-breaking procedures that are proven
7
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
~lVeneIS in promoting germination and lead to standardized interpretations in routine
the seed physiologist germination is the emergence of the radicle through the seed coat
to seed analysts germination is the emergence and development from the seed embryo of
essential structures which for the kind of seed in question are indicative of the ability
produce a normal plant under favorable condition (AOSA 1991) However according to
mllilti(Jlnal Seed Testing Association germination in a laboratory is the emergence and
lh~eloIDment from the seed being tested indicates the ability to develop into a normal plant
favorable conditions in the soil (1STA 1985)
separated from the parent plant seeds will undergo primary dormancy and sometimes
ondaly dormancy Dormancy prevents immediate germination and regulates the time
1dition and place that the germination will occur Dormancy helps seeds to survive under
nBrOnlLOle conditions They need favorable condition before they can germinate Copeland
McDonald (1995) agreed that seeds will undergo dormancy before able to germinate
bull m gthe period ofrest the seed is in a relatively inactive state and has a low metabolic rate
remain in this state until environmental conditions trigger the resumption of growth If
seeds germinate under natural conditions the individual has in a sense bet its life on the
ivonlble of environmental conditions for seedling establishment (Angevine and Chabot
8
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
Seed Storage
(1995) mentioned that suitable treatment of seeds produces satisfactory
amllnatlon of seeds and that good stands can be obtained when seeds are planted in clean
prepared seedbeds and given reasonable attention during the period of establishment
of certain species require special treatment for maximum gennination or breaking
~nanc--y (Copeland and McDonald 1995)
are seeds harvested and immediately planted without undergoing at least a brief
period Consequently the time of storage type of seed stored and storage
(temperature relative humidity and oxygen levels) influence seed vigor
ppellanQ and McDonald 1995)
affects the rate of donnancy loss in dry seeds (Roberts 1988) Copeland and
IliANIIIIIU (1995) stressed that the optimum temperature may be defined as the temperature
the greatest percentage of gennination within the shortest time The maximum
mure is governed by the temperature at which denaturation of proteins essential for
IIDIllUltilon occurs They also agreed that temperate-region seeds require lower temperature
do tropical region seeds and wild species have lower temperature requirements than do
rticalted plants This is because the responses of seed gennination to temperature
on a number of factors such as the species variety growing region quality of the
IIld duration of time from harvest Angevine amp Chabot (1979) observed that seed
to temperature play pivotal role and most important for synchronization of
IIIDau(lD with conditions suitable for seedling establishment Hanelt (1977) suggested that
9
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
is an ecological connection between seed storage characteristics and the habitat in which
Ullecumiddots is found
(1980) argued that the amount of moisture in the seeds is probably the most
IkB1tant factor influencing seed viability during storage The rate of deterioration increases
seed moisture content increases Too high moisture contents can cause rapid loss due to
growth on and in the seeds On the other hand too low moisture contents can accelerate
_ iCCIiltlltJn or cause hard-seededness in some kind Since the life of a seed and its span
revolves around its moisture content it is necessary to dry seeds to safe moisture
The seeds have to be stored at a critical moisture content which is usually little lower
the seed moisture content at harvest and lost viability with decrease in their moisture
humidity and temperature by far are the most important factor determining the
life of seeds The maintenance of moisture content during storage is a function of
humidity In sealed storage seed moisture content determines the relative humidity of
ClIrJrolunent in the containers (AOSA 1991)
1IOICl1ID2 to Roberts (1973) there are two categories of seed storage behavior orthodox and
pcrtnult Species with orthodox seeds can be maintained satisfactorily ex situ over a long
in appropriate environments but short term storage is usually the best that can be
with seeds which show recalcitrant seed storage behavior under well-defined and
l-CCBlUOIU~ environment
10
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
pllDl()n feature of seed deterioration is the loss of integrity of membrane cell As ageing
lJInssed membrane integrity within the seed at imbibition declined (Villers 1973) The
of sugars and electrolytes during the imbibition process increased This increase in
membranes permeability in seed as ageing progressed was due to seed impairment
was the first indication of quality decline (Abdul-Bald and Baker 1973 Mohd Lassim
CleClme in enzyme activity is a measurable symptom of ageing The general decrease in
activity in the seeds lowered its respiratory potential which in tum lowered both the
(A TP) and food supply to the geminating seed (App et aI 1971) In dry seeds the
state has been suggested to serve as a physical stabilizer and protector against
-rinlAt11JP reactions (Bernal-lugo and Leopold 1998)
maturation a seed is at its maximum potential vigor thereafter its quality begins to
The rate of deterioration is influenced by genetic make-up of the seeds and the
trorllllcmt particularly relative humidity (moisture) temperature to which the seeds are
_a High temperature and relative humidity are deleterious and shorten the life span of
Matured seeds would deteriorate and lose their vigor if left on the plants in the field and
exposed to prolonged wet humid weather prior to harvesting (Heydecker 1972)
deterioration is an irreversible process Once sees deterioration has occurred this
process cannot be reversed Simply stated low quality seeds cannot be made into
quality seed Some mechanism for preconditioning or treating seeds with fungicide
field emergence However these treatments only allow the optimum expression of
11
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
potential they do not alter the basic physiological quality of the seed Seed deterioration
varies among populations It is established that certain varieties exhibit less deterioration
others Even within a variety the storage potential of individual lots varies and even
a seed lot individual seeds have differing storage potential (Copeland and McDonald
deterioration is manifested by the progressive decline in germinability enzymatic as
as metabolic activities growth and vigor increase in susceptibility to adverse
mmmlents pests and diseases When inferior quality seeds are sown they emerged from
i1 at different rates resulting in erratic field stand of uneven maturity and poor yielding
1CR8SU1lg temperature and seed moisture therefore is an effective means of maintaining
quality in storage The influence of temperature and relative humidity on seed
cncX8tion is exerted right from storage through the development of seedlings and mature
High quality seeds are susceptible to deterioration when stored under adversed
MlitlIODS of high temperature and relative humidity rendering them worthless for planting
a period ofstorage (Harrington 1972)
bwnidity temperature and seed moisture are factors that influence the life span of
The effects of relative humidity (and its subsequent effect on seed moisture) and
_are are highly interdependent The capability to predict seed deterioration would be
valuable to seed companies and germplasm repositories since the loss of seed
_ be anticipated and seed stocks are replenished (Copeland and McDonald 1995)
12
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
deterioration is one of major problem in commercial cultivation in most countries since
ago This caused retention of few beneficial species to successfully cultivate out of its
Salunkhe et al (1985) mentioned that over US$ 800 million is lost annually due to
and as consequences of breakage and microorganism spoilage during
=non storage and shipping of seeds McDonald and Nelson (1986) stated that it is
that 25 (approximately US$ 500) of seed sales in 1984 was lost due to poor seed
13
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
CHAPTER THREE
MATERIAL AND METHOD
abdariffa seeds samples were collected taken from Gulf and Pacific Industries Sdn Bhd
8000 seeds were used for the entire research Only fresh matured and viable seeds
used The seeds were soaked with water and only the submerged seeds were taken out
rinsed Then the seeds were dried in ambient room for several hours After dried seeds
treated by dusting with Captan 50 WP to protect from fungi and also protected the seeds
rotting and improved germination (Agrawal 1980) The seeds were kept in an air-tight
jar before used in the subsequent experiments
evaluation was conducted to assess the quality of seeds used in this research These
consist of moisture content and germination The procedures were those prescribed by
IIOClab()DofOfficial Seeds Analysts (AOSA 1985)
replicates of 15 seeds each were placed into saucers and deliberated to get the green
Then the seeds were placed in an oven at 60degC for 48 hours After 48 hours the seeds
deliberated again to get the dry weight The percentages of moisture content of the seeds
calculated base on the Association of Official Seeds Analysts (AOSA 1985)
14
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15
content () = b - c x 100 b-a
where a =weight of empty saucer
b =weight of (a) + weight of seeds before placing in the oven
c =weight of ( a) + weight of seeds after drying in the oven
__(m test was based on the AOSA (1985) with 4 replications of 50 seeds each were
in each evaluation The seeds were put in containers filled with sand as growth medium
were planted in a unifonn layer of moist sand and then covered with thin layer of sand
to genninate if the radicles were about 5mm length emerged from the seed coat
total ofgerminated seeds were recorded between 4-10 days
_tatiC)Il () = ~ X 100 b
a=total ofgenninated seeds
b = total of seeds used
15