CHARACTERIZATION OF LOQUAT (Eriobotrya japonica Lindl.)
GENOTYPES CULTIVATED IN PAKISTAN ON THE BASIS OF
MORPHO-PHYSICAL TRAITS AND MOLECULAR MARKERS
Azhar Hussain
03-arid-374
Department of Horticulture Faculty of Crop and Food Sciences
Pir Mehr Ali Shah Arid Agriculture University
Rawalpindi, Pakistan 2009
CHARACTERIZATION OF LOQUAT (Eriobotrya japonica Lindl.)
GENOTYPES CULTIVATED IN PAKISTAN ON THE BASIS OF
MORPHO-PHYSICAL TRAITS AND MOLECULAR MARKERS
By
Azhar Hussain
03-arid-374
A thesis submitted in partial fulfillment of the requirements for the degree of
DOCTOR OF PHILOSOPHY
IN
HORTICULTURE
Department of Horticulture Faculty of Crop and Food Sciences
Pir Mehr Ali Shah Arid Agriculture University
Rawalpindi, Pakistan 2009
CERTIFICATION
I hereby undertake that this research is an original one and no part of this thesis
falls under plagiarism. If found otherwise, at any stage, I will be responsible for the
consequences.
Name: Azhar Hussain Signature: ______________
Registration No. : 03-arid-374 Date: ______________
Certified that the contents and form of thesis entitled “Characterization of loquat
(Eriobotrya japonica Lindl.) genotypes cultivated in Pakistan on the basis of morpho-
physical traits and molecular markers” submitted by “Mr. Azhar Hussain” has been
found satisfactory for requirement of the degree.
Supervisor: _______________________ (Dr. Nadeem Akhtar Abbasi) Member: ________________________ (Dr. Ishfaq Ahmad Hafiz) Member: ________________________ (Dr. S.M. Saqlan Naqvi) Member: __________________________ (Dr. Zahoor Ahmad) Chairman: ________________________ Dean: ____________________________ Director Advanced Studies: ______________________
Dedicated to my family
i
CONTENTS
Page
List of tables iv
List of figures vi
List of appendices vii
Acknowledgements viii
ABSTRACT x
1. INTRODUCTION 1
1.1 ORIGIN 1
1.2 HISTORY 1
1.3 PRESENT STATUS 2
1.4 CLIMATE AND SOIL 3
1.5 IMPORTANCE OF LOQUAT 3
1.5.1 Nutritional Value 3
1.5.2 Medicinal Importance 4
1.5.3 Economic Value 4
1.6 MAJOR PRODUCING COUNTRIES 5
1.7 RESEARCH GAPS 7
2. REVIEW OF LITERATURE 9
2.1 NEED OF CHARACTERIZATION 10
2.2 TRADITIONAL METHODS OF CHARACTERIZATION 11
2.2.1 Limitations of traditional methods 13
2.3 MOLECULAR MARKERS 14
2.3.1 Protein markers 15
2.3.2 DNA markers 16
2.3.2.1 RAPD Markers 21
2.3.2.2 Reproducibility of RAPD markers 24
2.3.2.3 Use of RAPD in loquat 26
2.3.2.4 Use of RAPD in other fruit plants 29
ii
2.3.2.5 Molecular markers – supplement to conventional research 37
2.4 EVOLUTION OF VARIETIES 38
2.5 STATUS OF GERMPLASM RESOURCES OF LOQUAT 39
2.6 A BRIEF DESCRIPTION OF SOME LOQUAT
CULTIVARS
40
2.7 TRENDS IN LOQUAT RESEARCH 50
2.7.1 China 50
2.7.2 Spain 51
2.7.3 Turkey 53
3. MATERIALS AND METHODS 54
3.1 SURVEY OF THE LOQUAT GROWING AREAS 54
3.1.1 Selection of sites 54
3.1.2 Selection and tagging of plants 57
3.1.2.1 Kalar Kahar 57
3.1.2.2 Choa Saiden Shah 58
3.1.2.3 Chhattar 58
3.1.2.4 Tret 58
3.1.2.5 Hasan Abdal and Wah 59
3.1.2.6 Hari Pur 59
3.1.2.7 Mardan 59
3.1.2.8 Takht Bhai 60
3.2 EVALUATION OF GENOTYPES ON THE BASIS OF
MORPHO-PHYSICAL CHARACTERISTICS
60
3.2.1 Morphological characteristics 60
3.2.2 Physical traits 61
3.2.3 Data Analysis 64
3.3 DNA POLYMORPHISM ANALYSIS 64
3..3.1 DNA Extraction 65
3.3.2 Agarose gel electrophoresis 66
3.3.3 Polymerase chain reaction 66
3.3.4 RAPD Data analyses 68
iii
4. RESULTS AND DISCUSSION 69
4.1 CHARACTERIZATION OF LOQUAT GENOTYPES ON
THE BASIS OF MORPHO-PHYSICAL TRAITS
70
4.1.1 Kalar Kahar 70
4.1.2 Choa Saiden Shah 80
4.1.3 Chhattar 89
4.1.4 Tret 99
4.1.5 Hasan Abdal and Wah 110
4.1.6 Hari Pur 117
4.1.7 Mardan 127
4.1.8 Takht Bhai 137
4.1.9 Correlation among some physical traits of loquat genotypes 152
4.2 CHARACTERIZATION OF LOQUAT GENOTYPES ON
THE BASIS OF MOLECULAR MARKERS
158
4.2.1 RAPD Analysis 159
4.2.2 Cluster Analysis Based on RAPD Markers 161
SUMMARY 171
LITERATURE CITED 176
iv
List of Tables
Table No. Page
1 Geographical locations of the selected loquat sites 56
2 Ten base pair primers used for the DNA amplification of
loquat
67
3 General appearance of the loquat plants of 5 genotypes at Kalar
Kahar
72
4 Fruit and seed morphology of 5 loquat genotypes at Kalar Kahar 72
5 Fruit characteristics of 5 loquat genotypes at Kalar kahar 74
6 Seed characteristics of 5 loquat genotypes at Kalar Kahar 77
7 Leaf characteristics of 5 loquat genotypes at Kalar Kahar 78
8 Floral characteristics of 5 loquat genotypes at Kalar Kahar 79
9 General appearance of the loquat plants of 3 genotypes at Choa
Saiden Shah
81
10 Fruit and seed morphology of 3 loquat genotypes at Choa Saiden
Shah
81
11 Fruit characteristics of 3 loquat genotypes at Choa Saiden Shah 83
12 Seed characteristics of 3 loquat genotypes at Choa Saiden Shah 87
13 Leaf characteristics of 3 loquat genotypes at Choa Saiden Shah 87
14 Floral characteristics of 3 loquat genotypes at Choa Saiden Shah 88
15 General appearance of the loquat plants of 3 genotypes at Chhattar 90
16 Fruit and seed morphology of 3 loquat genotypes at Chhattar 90
17 Fruit characteristics of 3 loquat genotypes at Chhattar 93
18 Seed characteristics of 3 loquat genotypes at Chhattar 96
19 Leaf characteristics of 3 loquat genotypes at Chhattar 97
20 Floral characteristics of 3 loquat genotypes at Chhattar 98
21 General appearance of the loquat plants of 5 genotypes at Tret 100
22 Fruit and seed morphology of 5 loquat genotypes at Tret 100
23 Fruit characteristics of 5 loquat genotypes at Tret 104
24 Seed characteristics of 5 loquat genotypes at Tret 108
25 Leaf characteristics of 5 loquat genotypes at Tret 108
v
26 Floral characteristics of 5 loquat genotypes at Tret 109
27 General appearance of the loquat plants of 5 genotypes at Hasan
Abdal & Wah
111
28 Fruit and seed morphology of 5 loquat genotypes at Hasan Abdal
& Wah
111
29 Fruit characteristics of 5 loquat genotypes at Hasan Abdal & Wah 114
30 Seed characteristics of 5 loquat genotypes at Hasan Abdal & Wah 118
31 Leaf characteristics of 5 loquat genotypes at Hasan Abdal & Wah 119
32 Floral characteristics of 5 loquat genotypes at Hasan Abdal & Wah 120
33 General appearance of the loquat plants of 3 genotypes at Hari Pur 122
34 Fruit and seed morphology of 3 loquat genotypes at Hari Pur 122
35 Fruit characteristics of 3 loquat genotypes at Hari Pur 125
36 Seed characteristics of 3 loquat genotypes at Hari Pur 128
37 Leaf characteristics of 3 loquat genotypes at Hari Pur 129
38 Floral characteristics of 3 loquat genotypes at Hari Pur 130
39 General appearance of the loquat plants of 3 genotypes at Mardan 131
40 Fruit and seed morphology of 3 loquat genotypes at Mardan 131
41 Fruit characteristics of 3 loquat genotypes at Mardan 134
42 Seed characteristics of 3 loquat genotypes at Mardan 138
43 Leaf characteristics of 15 loquat genotypes at Mardan 139
44 Floral characteristics of 15 loquat genotypes at Mardan 140
45 General appearance of the loquat plants of 15 genotypes at Takht
Bhai
142
46 Fruit and seed morphology of 15 loquat genotypes at Takht Bhai 143
47 Fruit characteristics of 15 loquat genotypes at Takht Bhai 146
48 Seed characteristics of 15 loquat genotypes at Takht Bhai 153
49 Leaf characteristics of 15 loquat genotypes at Takht Bhai 154
50 Floral characteristics of 15 loquat genotypes at Takht Bhai 155
51 Correlations among some physical traits of 42 loquat genotypes 156
52 Polymorphism revealed by different RAPD primers 160
vi
List of Figures
Fig. No Page
1 Map of Pakistan showing main loquat growing areas 55
2 RAPD pattern of loquat genotypes obtained with Primer A-02 165
3 RAPD pattern of loquat genotypes obtained with Primer C-02 166
4 RAPD pattern of loquat genotypes obtained with Primer C-05 167
5 RAPD pattern of loquat genotypes obtained with Primer C-07 168
6 RAPD pattern of loquat genotypes obtained with Primer C-19 169
7 Clustering pattern of loquat genotypes based on RAPD markers 170
vii
List of Appendices
Appendix No. Page
1 List of loquat genotypes included in study 208
2 Area and production of loquat in different countries 209
3 Area and production of loquat in Pakistan during last 5
years
210
4 Area and production of loquat in provinces of Pakistan 211
5 Loquat germplasm resources in different countries 212
6 Binary matrix of 42 loquat genotypes as obtained by
Primer GL DecaemerA-02
213
7 Binary matrix of 42 loquat genotypes as obtained by
Primer GL DecaemerC-02
214
8 Binary matrix of 42 loquat genotypes as obtained by
Primer GL DecaemerC-05
215
9 Binary matrix of 42 loquat genotypes as obtained by
Primer GL DecaemerC-07
216
10 Binary matrix of 42 loquat genotypes as obtained by
Primer GL DecaemerC-19
217
viii
ACKNOWLEDGMENTS
Thanks to Almighty Allah who created all the things for the benefit of mankind.
Praise for His last Prophet (Peace be upon him) who spread the message to serve the
humanity and conserve the plants.
I express my deep sense of admiration to my supervisor, Prof. Dr. Nadeem A.
Abbasi, Chairman Department of Horticulture for his inspiration, support and
encouragement throughout this research project. I am thankful to him for his professional
guidance and constructive criticism. Working under his supervision as a doctorate student
has been a glorious opportunity for me.
I am thankful to Dr. Ishfaq Ahmad Hafiz, Associate Professor Department of
Horticulture for his technical direction and moral support. I always found him available
whenever I needed his help. I am also grateful to Prof. Dr. S.M. Saqlan Naqvi, Chairman
Department of Biochemistry for his technical supervision and providing me the laboratory
facilities. He gave me confidence to face the problems and find their solutions. Thanks are
also due to Dr. Zahoor Ahmad, Senior Director, Crop Research Institute (National
Agricultural Research Centre Islamabad), who always encouraged, and appreciated me in
accepting and resolving the challenges while conduct of this research.
I am thankful to the loquat growers of all the experimental sites, especially Mr.
Ikramullah Khan (Mardan) and Hafiz Masoud (Kalar Kahar) for their cooperation. Thanks
to all my friends, Dr. Shabbir and Dr. Rabbani for their motivation to win the scholarship,
Dr. Zeeshan, Dr. Sajid and Dr. Tariq (Biochemistry department) for their technical help,
Mr. Sajjad Hussain Shah and Raja Shahid for their support during field survey and data
collection, Mr. Touqeer Ahmad (Lecturer) for providing better environment in the
laboratory, Mr. Attiq and Mr. Javed for their long time company in the lab and all other
friends for their cooperation and good wishes for me.
Different departments out side the university also facilitated me in one way or the
other for accomplishment of this work. I cannot forget the help extended by the staff of
Agricultural Extension Choa Saiden Shah, Agricultural Extension Taxila, Hill Fruit
ix
Research Station Murree, Barani Agricultural Research Institute Chakwal and On-Farm
Water Management Project Rawalpindi,
Thanks are also extended to the Higher Education Commission Pakistan for
awarding me Ph.D. Indigenous Fellowship, which helped me in conduct of this study.
I am also thankful to my family members, especially my mother for her generous
prayers and my wife for her cooperation and patience throughout the hectic Ph.D.
schedule.
AZHAR HUSSAIN
x
ABSTRACT
Loquat (Eriobotrya japonica Lindl.) is an important but ignored fruit crop of
Pakistan for which no research work has ever been reported previously inside the country.
There is no standard or identified loquat cultivar available to the growers for cultivation in
the loquat growing pockets of Pakistan. Generally, the farmers grow their orchards
through seeds. As a result, most of the loquat orchards do not possess the plants with
uniform fruit characters and fruit is not of good quality. Previously no work has been
reported regarding description of the loquat genotypes in Pakistan.
The present study was, therefore, carried out to evaluate and characterize the
available genotypes in the main loquat growing areas of Pakistan and to determine the
genetic diversity among these genotypes. For this purpose, 9 sites were selected in the
main loquat growing areas of Pakistan. Forty two genotypes were identified, which were
compared on the basis of morpho-physical traits. Significant differences were observed
with reference to various characteristics among different genotypes. Fruit weight of the
genotypes ranged from 9.54 g (in HW4) to 47.84 g (in TB15). Range of flesh seed ratio
was from 1.67 (in HW5) to 3.05 (in TB8). Minimum yield per tree was recorded as 25.85
kg (in TB15), while it was maximum (89.87 kg) in TB7. Correlations among some traits
were also observed.
Moreover, RAPD analyses of the genotypes were performed. Five RAPD
primers gave reproducible results and generated 47 polymorphic bands. According
to the dandrogram, two main groups of the loquat genotypes were identified with a linkage
distance of 33%. For most of the locations, grouping of the genotypes was in accordance
with the geographical locations. Out of the three genotypes from Mardan, one falls under
xi
the first group and the other two under the second group. The maximum number of
genotypes (15) was identified at Takht Bhai, two of them belonged to the first group and
13 to the second group.
Genotypes with good characteristics i.e. better yield, higher fruit size and weight,
less number of seeds per fruit and higher flesh seed ratio can be recommended for further
multiplication and introduction to the other loquat growing areas which would increase the
income of farming community. The study also recommends establishing germplasm units
in Punjab and NWFP and pooling all these genotypes for future strategies and breeding
programs including selection, introduction, hybridization and mutation breeding. The
present study would also be helpful for the documentation, management, and conservation
of the loquat genetic resources of Pakistan.
1
Chapter 1
INTRODUCTION
Loquat (Eriobotrya japonica Lindl.) is an important sub-tropical fruit tree
belonging to the family Rosaceae, subfamily Pomoideae. It blooms in fall (Razeto
et al., 2003) hence is a good source of nector. It is under cultivation in many
countries. In China, it is called “Pipa” or“Luju”. In the United States it is also
known as Japanese medlar or Japanese plum, in France as neflier du Japon, in
Spain as Nispero in Italy as nespola, in Portugal as ameixa do Japao and in
Germany as Japonische Mispel (Lin et al., 2007).
1.1 Origin
The Dadu River Valley of China is believed to be the original home of the
genus Eriobotrya. Most authors consider that the loquat species originated in
China (Campbell and Malo, 1986; Zhang et al., 1993; Polat and Caliskan, 2007;
Zheng, 2007; Huang et al., 2007), from where it spread to other countries of the
world.
1.2 History
Loquat has been under cultivation for over 2000 years (Lin et al., 2007)
since the Chinese Han dynasty (Zhu et al., 2007). The loquat in Japan was
introduced from China in the ancient period and its gardening in Japan was done as
early as 1180 (Ichinose, 1995). But this ancient fruit has become commercialized
in recent times on a large scale (Janick, 2007).
2
Loquat cultivation in Eastern Asia is very ancient while the crop’s spread to
Europe took place more recently, in 1784 when it was established in the Botanical
Gardens of Paris. From here, the loquat made its way to the Mediterranean region
and afterward reached Florida and California from Europe and Japan respectively
(Vilanova et al., 2001).
From Japan, it was introduced to Europe as an ornamental tree in the 18th
century. But later during the 19th century, selections of cultivars with larger fruits
were made for fruit production (Badenes et al., 2000).
1.3 Present status
Loquat is cultivated mainly in China, Japan, Pakistan, India, Madagascar,
Mauritius Island, Reunion Island, the Mediterranean countries (Turkey, Spain,
Italy, Greece and Israel), United States (largely California and Florida), Venezuela,
Brazil and Australia (Badenes et al., 2000; Vilanova et al., 2001; Li et al., 2007).
So far, it has been grown in more than 30 countries of the world (Feng et al.,
2007). Loquat is becoming an important industry in China as well as Spain, Japan,
India, Pakistan and Turkey (Janick, 2007). In Pakistan it is mostly consumed in
the local or short distant markets (Hussain et al., 2007).
World loquat fruit production is 549,220 tonnes, China (460,000
tonnes) and Spain (43,300 tonnes) being the main producers followed
by India, Pakistan and Japan (Lin, 2007, Lin et al., 2007). In Pakistan,
its production is 10,479 tonnes, 98 % of which comes from the North
Western Frontier Province (NWFP) and Punjab (GOP, 2008). It is being
3
grown in Punjab at Chattar, Tret, Kalar Kahar, Choa Saiden Shah, Wah
and Hasan Abdal, while in NWFP it is cultivated in Mardan region,
Peshawar and Hari Pur (Hussain et al., 2007).
1.4 Climate and soil
Loquat has adapted well to the Mediterranean climate and produced in the
same areas where citrus is cultivated (Badenes et al., 2000). However, it has more
specific environmental requirements than citrus (Caballero and Fernandez, 2003;
Durgac et al., 2006).
Generally, the loquat tree is well adapted to almost all soils that have good
drainage and hence grows equally well in acidic as well as in alkaline soils. The
tree is cold tolerant to -10°C but the fruits freeze at low temperature of about -3°C
(Lin et al., 2007). To establish a loquat orchard, winter temperature should be
higher than -3°C and summer temperature not above 35°C (Lin, 2007).
Loquat has formed a variety of ecological types in different zones over the
course of its cultivation and acclimatization. Generally, it can be found in maritime
climates between the latitudes 20 and 35 north and south (Vilanova et al., 2001)
but can be grown up to the latitude 45 (Lin et al., 1999; Polat and Caliskan, 2007).
1.5 Importance of loquat
1.5.1 Nutritional value
Mainly loquat is consumed as fresh fruit. Besides having a sweet
taste and juicy texture, it is very nutritious. According to Karadeniz (2003),
4
it contains vitamins (A, B, and C), minerals (phosphorus and calcium) and
sugars.
1.5.2 Medicinal importance
Fruit and leaves of loquat have been considered to have high medicinal
value (Wee and Hsuan, 1992). ‘Feitai’, a compound formula, consisting of a
number of herbs including loquat leaves, is used in China as a folk medication for
treating the patients with pulmonary tuberculosis (Zhang et al., 2004). Loquat
leaves are known to have many physiological actions such as expectorant and anti-
inflammatory (Hamada et al., 2004) and are used to treat skin diseases and to
relieve pain (Sakuramata et al., 2004, Nishioka et al., 2002), inflammation
(Nishioka et al., 2002) and coughing (Sakuramata et al., 2004). Loquat leaves have
ursolic acid and oleanolic acid both having hypoglycaemic and antihyperlipidaemic
properties (Saliba et al., 2004). Leaves also contain anti-tumor agents (Ito et al.,
2002) and furthermore have the anti-diabetic properties (Sakuramata et al., 2004).
The loquat seeds contain the unsaturated fatty acids linolenic and linoleic acids and
the sterol beta-sitosterol, which may contribute to the improvement of liver
function (Nishioka et al., 2002). Loquat seed extract has an inhibitory effect on
liver disorders (Hamada et al., 2004). It also has the anti-inflammatory effects
(Takuma et al., 2005).
1.5.3 Economic value
World loquat production is 549,220 tonnes while area is 131,260
hectares. Pakistan produces 10,479 tonnes from an area of 1501 hectares
5
(GOP, 2008) while a very little amount is exported to the Middle Eastern
countries mainly Dubai (Khan, 2003).
Previous years’ statistics show a gradual increase in the loquat area
as well as production in Pakistan. There is great potential of increase in the
loquat area and hence production of the country. Cultivation of superior
genotypes may further increase the production, hence increasing its
availability for the home market as well as for export (Hussain et al.,
2007).
Loquat fruit develops during winter and ripens at early spring. Due
to its unusual phenology, it reaches the market before any other fruit of the
spring season (Cuevas et al., 2007). In Pakistan, loquat fruit becomes
available in the months of March / April when no other fresh fruit is
available in the market, thus filling a gap between oranges and the first
stone fruits of the season. As it is the first fruit of the year, it is very popular and
sells at a best price (Khan, 2003). Since it gives flowers in the autumn, it is a good
source of nectar when other resources are scarce (Merino and Nogueras, 2003).
1.6 Major producing countries
Before the foundation of modern China, loquat was an underutilized tree
species. After the commencement of loquat breeding, several scientists in China
studied loquat resources. Loquat breeding has been carried out regularly by means
of introduction, selection and hybridization (Zheng, 2007). Now China is at the top
in terms of area (120,000 hectares) as well as the production (460,000 tonnes) of
6
loquat (Lin et al., 2007; Feng et al., 2007; Huang et al., 2007). From the 1970s,
loquat production in China witnessed a fast increase from 2000 hectares to 26,000
hectares in 1995 and to 120,000 hectares in 2005.
Spain is the second largest loquat producer in the world and the first
exporting country. It accounts for 84 % of worldwide loquat trade, with the major
destination being European Union countries: Italy, France and Portugal and exports
36 to 47 % of the total production (Llacer and Soler, 2001; Caballero and
Fernandez, 2003; MAPA, 2004; Soler et al., 2007; Canete et al., 2007; Hueso et
al., 2007). Spain leads in loquat production in the Mediterranean region with about
3,700 hectares of orchards, which give a production of 45,000 tonnes annually. In
Spain, loquat production increased considerably in the last 20 years from 18,000
tonnes in 1985 to 45,000 tonnes in 2004 (MAPA, 2004; Canete et al., 2007).
Turkey is the third important producer of loquat with a production of 12000
tonnes from an area of 820 hectares (Lin, 2007). Area under loquat in Turkey is
less as compared to Japan (2,420 hectares) or Pakistan (1501 hectares). A rapid
increase has been observed here after 1980. The total production was only 3000
tonnes in 1980 which increased to 9000 tonnes by 1990 and 12000 tonnes by 2003
(Polat and Caliskan, 2007; Karadeniz and Senyurt, 2007).
Before World War II, Japan was the largest loquat producing country. After
the war, area under loquat in Japan gradually reduced because development of food
crops became more essential and cultivation of loquat was too labour consuming.
Now Japan has a production of 10,240 tonnes from an area of 2420 hectares (Lin et
al., 2007; Lin, 2007).
7
1.7 Research gaps
There is no standard or identified loquat cultivar available to the
growers for cultivation in the loquat growing pockets of Pakistan. As a
result, most of the loquat orchards do not possess the plants with uniform
fruit characteristics. Most of the orchards have seedling trees with variable
performance owing to heterozygosity and cross-pollination. Therefore, the
loquat growers face difficulties during harvesting and marketing due to
variation in fruit size and quality.
A lot of trees with good fruit size and better yield can be identified for
which no effort in Pakistan has ever been made. Hence there is scope of selection
of varieties with good characters i.e. big fruit size, less number of seeds, more flesh
and prolific bearing (Khan, 2003). A number of loquat genotypes are there in
the loquat growing areas of Pakistan, while previously no work has been
reported regarding the description of these genotypes (Hussain et al.,
2007).
The present study is aimed to evaluate and characterize the available
genotypes in the main loquat growing areas of Pakistan, and to identify the
genotypes with better characteristics for further propagation and uniform
plantation. It will also be helpful to conserve the quality plants and provide
a documentation of the local loquat germplasm with reference to their
similarity and diversity.
Since no molecular information is available regarding loquat cultivars
grown in Pakistan, the study is aimed at determining the genetic relationships and
8
diversity among the different genotypes growing in different parts of the country.
The information obtained from this work may be useful for the management of the
genotypes studied. Cultivation of superior genotypes may help to increase
the production, hence increasing its availability for the home market as
well as for export (Hussain et al., 2007). This study will also provide the
guidelines for any future research on loquat.
9
Chapter 2
REVIEW OF LITERATURE
Information of genetic diversity amongst the adapted cultivars or selected
breeding materials has a major impact on the crop improvement. It can be acquired
from the pedigree analysis, morphological characters or using molecular markers
(Pejic et al., 1998). This knowledge is helpful in the management of gene bank and
breeding experiments like labeling of germplasm, identification and exclusion of
duplicates in the gene stock and establishment of core collection. Another
application of the knowledge of genetic diversity is in sorting of the populations
for genome mapping (Kaga et al., 1996; Nisar et al., 2007).
Genetic diversity between and within plant populations results from a
combination of physical remoteness, population size, type of mating system
(selfing or outcrossing), mode of dispersal of seed and pollen, and speed of gene
flow (Loveless and Hamrick, 1984, Mekuria et al., 2002). Species that are largely
out crossing show lower inter-population and higher intra-population difference in
genetic variation compared to the species where self-fertilization predominate
(Maguire and Sedgley, 1997).
New improvements in molecular biology have made the plants’ DNA
fingerprinting feasible and a number of techniques are available today to
characterize DNA polymorphism. Quite a lot of of these techniques have been used
in phylogenic studies, genetic diversity analysis and varietal characterization of
plants (Williams et al., 2004).
10
2.1 Need of characterization
Exact and quick identification of the cultivars is particularly important in
vegetatively propagated plants for the purpose of practical breeding as well as for
the protection of proprietary rights (Nicese et al., 1998). The varietal identification
is also important for the documentation of genetic resources and for the protection
of breeders’ benefit (Selbach and Cavalli-Molina, 2000).
Assessment of genetic diversity to recognize groups with similar genotypes
is very important to conserve, evaluate and utilize the genetic resources, for
studying the diversity of the germplasm as potential basis of genes that may be
capable to improve the performance of cultivars, and for determining the
distinctness and uniqueness of the phenotypic and genetic formation of genotypes
with the purpose of protecting the intellectual property rights of the breeder
(Subudhi et al. 2002; Nemera et al., 2006). Diversity studies would also be
desirable for the purpose of better management and conservation of the genetic
resources and for planning the breeding strategies (Badenes et al., 2000).
Due to lack of even the basic set of information about loquat germplasm in
Pakistan, investigations on the number of genotypes and their geographical
distributions as well as the potentially useful trees with desirable traits are
necessary. Management of germplasm and conservation of genetic resources can
be carried out after detailed characterization of plant material (Badenes et al.,
2004). Detailed studies on genetic diversity can be accomplished by studying the
morphological traits or by utilizing teh marker systems such as allozymes, RFLPs,
AFLPs RAPDs, or SSRs (Nemera et al., 2006). Collection and conservation of
11
loquat germplasm is essential for breeding purposes as well as for saving the
germplasm, which is at the edge of extermination (Wang et al., 2007).
The main problem associated with the loquat plant material is the loss of
genetic diversity, as the local cultivars not meeting the market requirements have
been replaced by other prominent cultivars with good characteristics. In order to
resolve this problem, there is a need to carry out two complementary research
projects, firstly to collect, conserve and characterize the loquat genetic resources
and secondly to initiate loquat breeding programmes so that new cultivars can be
obtained that could broaden the range of available cultivars adapted to the market
demands as proposed by Llacer et al. (2003).
2.2 Traditional methods of characterization
Conventionally, genetic diversity has been assessed on the basis of
dissimilarity in morphological and agronomic characters or on ancestry
information for the different crops (Sneller et al., 1997; Bernard et al., 1998; Liu et
al., 2004). Before the progress in the field of biotechnology, only the
morphological and physical characteristics were taken under consideration while
characterizing different cultivars. Same is the case with loquat. The basis of
identification and characterization of loquat cultivars has been the morphological
and pomological traits. Characteristics like leaf blade length and width, shape of
leaf blade, thickness and distribution of the lateral and middle shoots and size and
shape of flower clusters were used to distinguish cultivars. Similarly, fruit size,
colour of fruit and flesh, fruit shape, shape of stalk end and that of apex and calyx
12
cavity were considered important to distinguish different loquat cultivars (Badenes
et al., 2000).
In, India, 15 nut and kernel traits were used to assess the genetic divergence
among 229 naturally growing seedling trees of Persian walnut in four districts of
Himachal Pradesh. The parameters used for this purpose included nut weight, nut
width, nut height, nut thickness, index of roundness, shell thickness, kernel weight,
kernel width, kernel thickness, kernel percentage, fat percentage and protein
percentage (Sharma and Sharma, 2001).
Several researchers used the plant characteristics related to flowers, fruits
and leaves to illustrate and characterize different varieties of mandarin and its
hybrids (Domingues et al., 1999; Koehler-Santos et al., 2003).
Traditional methods of characterization were not common only in the past
but their significance is still recognized and the cultivars are illustrated through the
traits like leaf blade length, leaf blade width, seed weight, seed number, flesh /
seed ratio, fruit weight, size, shape and colour (Nandini and Chikkadevaiah, 2005;
Durgac et al., 2006; Hussain et al., 2007).
Hatay province is situated in the Eastern Mediterranean area in Turkey.
Generally, the loquat plants are mixed with other fruit trees. Traditionally, loquat
cultivation in the Hatay basin comprises the isolated trees found in gardens, small
plantations or family orchards. A study has recently been conducted to select high-
quality genetic resources of loquat on the basis of conventional research
parameters. Loquat accessions were studied in terms of fruit characteristics such as
13
fruit size, skin colour, number of seeds, flesh / seed ratio and total soluble solids.
As a result, , 13 genotypes have been selected among these populations for future
breeding work (Polat, 2007).
In past, all the important cultivars have been described on the basis of their
morphology and taxonomy through conventional research. Plant breeders used to
make the selections of breeding material on the basis of morphological traits that
were readily recognizable. Although the morphological characterization is a time
consuming process and is influenced by the environment, it is still considered to be
a practical way of making progress in the process of germplasm evaluation
(Nemera et al., 2006).
2.2.1 Limitations of traditional methods
Although conventional taxonomic approach has been traditionally
exercised for the varietal identification and can give a unique identification of the
cultivated varieties, it is not well appropriate to provide reliably judicious
identifications. It is not possible to find out how completely the genome has been
sampled by morphological description (Nandini and Chikkadevaiah, 2005).
Though the selection of breeding material on the basis of morphological
characteristics has been an effective method, morphological comparisons may have
limitations, including the influence of the environment or management practices
(Gepts 1993; Nemera et al., 2006).
The conventional methods for characterization and evaluation of genetic
variability in perennial fruit crop species, based on morphological and
14
physiological studies, are both time consuming and influenced by the environment
(Nicese et al., 1998). It is, therefore, difficult to distinguish genotypes just on the
basis of their external morphology. Furthermore, these phenotypic characters are
normally influenced not only by the environmental factors but also by the growth
stage of the plant (Baranek et al., 2006).
The conventional approach to characterize the cultivars in fruit tree species
on the basis of phenotypic observations is slow due to the long generation time.
Therefore, new methods based on studies at the DNA level should be incorporated
into fruit breeding programs in order to speed up and optimize genotype
fingerprinting and to study genetic associations among cultivars (Wunsch and
Hormaza, 2002; Shiran et al., 2007).
2.3 Molecular markers
Molecular markers are very rapidly being adopted by the researchers all
over the world for the crop improvement and are the appropriate and valuable tools
for basic and applied studies dealing with the biological mechanism in agricultural
production systems (Jones et al., 1997; Mohan et al., 1997). These tools are more
dependable than the phenotypic observations for the purpose of evaluating the
variations and examining the genetic stability (Leroy et al., 2000). These molecular
techniques differ in principle, application and amount of polymorphism observed
and in time requirements (Vilanova et al., 2001; Naghavi et al., 2004).
Molecular markers put forward an efficient tool for cultivars fingerprinting,
assessment of genetic resemblance and relationships (Vilanova et al., 2001) and
15
provide the best estimation of genetic diversity as they are independent of the
confusing effects of the environmental factors (Naghavi et al., 2004). We can
detect them in all tissues and at all stages of development (Badenes et al., 2004).
Molecular markers are exercised in different fields of genetics such as
genetic mapping, genome organization, characterization and identification of plant
cultivars. They are very appropriate means for the characterization of genotypes in
the gene banks (Raddova et al., 2003).
Use of molecular markers is even more important for the perennial and
recalcitrant crops, where progress in crop improvement is frequently hindered by
its long generation time (Upadhyay et al., 2004; Shiran et al., 2007).
Molecular markers such as isozymes (Lin, 1990), RAPD (Vilanova et al.,
2001; Pan et al., 2002; Badenes et al., 2004; Luo et al., 2007), SSR (Soriano et al.,
2005; Gisbert et al., 2007b) and AFLP (Feng et al., 2007 ) have been used for the
genetic diversity studies in the perspective of loquat genepools. Molecular markers
may be protein in nature or DNA based.
2.3.1 Protein markers
Protein markers, including structural proteins, seed storage proteins, and
isozymes were among the first collection of molecular markers exploited for the
assessment of genetic diversity and for the development of genetic linkage map.
Isozymes were the earliest genetic markers exercised in genotype fingerprinting
and have been applied in a number of fruit species. However, their effectiveness
has been restricted due to the small number of isozymes systems existing, the low
16
level of polymorphism obtained, and the influence of the environmental factors
(Khadari et al., 2005).
Although, isozymes supplied useful information (Chung and Ko, 1995;
Chevreau et al., 1997) but have some drawbacks like the limited number of
polymorphisms detected between close genotypes and inconsistency due to the
physiological stage (Oliveira et al., 1999).
2.3.2 DNA markers
Molecular markers that detect variation at the DNA level overcome most of
the limitations of biochemical and morphological markers. As confirmed by their
use in a variety of plant species, molecular markers are most appropriate for
assessment of genetic diversity and identification of varieties (Upadhyay et al.,
2004).
DNA markers basically detect differences in genetic makeup; in other
words, they are based on polymorphism in DNA sequences carried by different
individuals (Samec, 1993).
DNA markers are a more stable and useful substitute to isoenzymes. These
markers can more efficiently be used to determine the genetic diversity of
collections. In reality, many molecular markers are presently being used to
examine the genetic diversity of different species (Colombo et al., 2000).
Development of molecular markers associated with the traits of interest has the
advantage that desirable genotype of the plants can be selected in the early stage of
development (Gisbert et al., 2007a).
17
DNA markers are very valuable fundamental tool that plant breeders use
for the pedigree analysis, cultivar identification, and assessing genetic diversity.
DNA-based markers provide an opportunity for the genetic characterization that
allows the direct comparison of different genetic material without being influenced
by the environmental conditions (Weising et al., 1995; Nicese et al., 1998).
Use of analytical techniques based on DNA amplification for the study of
genetic diversity and relationships within the collections of genetic resources of
plant material is very common. Since the evaluation based on the morphological
characteristics is very time consuming and it may take several years in case of fruit
trees and other perennial plants, DNA based methods are more useful and more
economical (Raddova et al., 2003; Shiran et al., 2007). They can be used more
reliably than the phenotypic observations to evaluate the variations and to observe
the genetic stability. This fact highlights the need for alternative and
unconventional methods of ultimate detection based on molecular techniques like
RAPD or AFLP (Gaafar and Saker, 2006).
RFLP (restriction fragment length polymorphism) analysis involves
extensive labour, is highly expensive and time consuming, and is therefore
unfeasible when analyzing huge germplasm samples (Sarkhosh et al., 2006). It has
the limitations due to radioactive needs and complex methodology, in addition to
the larger genomic DNA requirement (Gaafar and Saker, 2006). Compared to
RFLP markers, RAPD markers can generate markers more rapidly (Selbach and
Cavalli-Molina, 2000) and data can be produced faster with less labour than RFLP
and microsatellites (Modgil et al., 2005).
18
PCR (polymerase chain reaction) based assays are considered to meet both
the genetic and technical requirements for the characterization of animal and plant
genetic resources (Powell et al., 1995). PCR based methods require lesser amounts
of genomic DNA, are non-radioactive, relatively low costing, and can be
developed rapidly (Al-Humaid and Motawei, 2004). The emergence of new
polymerase chain reaction (PCR) based molecular markers, such as randomly
amplified polymorphic DNA (RAPD), amplified fragment length polymorphisms
(AFLPs), and simple sequence repeats (SSR) has produced the opportunity for
excellent level genetic characterization of germplasm collections because they are
very much polymorphic and are not readily affected by the environmental
conditions (Geuna et al., 2003; Hokanson et al., 2001; Shiran et al., 2007).
In this group, RAPD (random amplified polymorphic DNA) is the method
which is most commonly used so far and was introduced by Williams et al.,
(1990). After a few years, the somewhat similar ISSR (inter simple sequence
repeats) (Zietkiewicz et al., 1994) and to some extent more technically demanding
AFLP (amplified fragment length polymorphism) (Vos et al., 1995) were
established.
Later on, STMS (sequence tagged microsatellite sites), which are based on
the micro satellite DNA loci with tandem repeats of one to six nucleotides became
increasingly popular for the population analysis. These loci are analyzed with PCR,
using sequencing information to develop the necessary primers. Variation at the
micro satellite loci, also known as simple sequence repeats (SSR), is generally
19
studied at all the identified loci separately, and can then be regarded as co-
dominantly inherited (Nybom, 2004).
Polymerase chain reaction (PCR) derived markers obtained with non
specific primers have become remarkably popular because they do not require
sequence information for the target species. As a result, these methods are
particularly suited to the circumstances where little or no research on molecular
genetics has been accomplished previously (Nybom, 2004).
SSRs and ISSRs are based on micro satellite and flanking sequences. These
have been observed to detect very high levels of polymorphism (Fahima et al.,
1998). However, previous knowledge about the genome is required before SSR
markers can be exploited to their maximum potential (Sarkhosh et al., 2006).
There is a lack of information regarding the minor fruit species at
molecular level; therefore, sequences flanking microsatellites from these species
are not available. Due to all these facts, RAPD markers are the most suitable
choice for characterization of the minor fruit species (Badenes et al., 2004).
AFLP markers are one of the most modern innovations in the genetic
marker technologies. These are based on RFLPs and RAPDs (Vos et al., 1995).
Both, AFLP and RAPDs can generate valuable information in loquat, but not all
markers are suitable for all purposes. The cost of generating the marker
information and the marker resolution are important considerations while selecting
a marker system. AFLP markers are more difficult to obtain and they cannot be
applied on a routine basis for fingerprinting. In convenient terms RAPD
20
technology is very helpful in the management of loquat germplasm (Vilanova et
al., 2001).
By using the AFLP (amplified fragment length polymorphism) practice a
large number of polymorphic DNA markers can be detected in a relatively short
time and is therefore a useful technique when high throughput is preferred (Vos et
al., 1995). These markers are more reliable and reproducible than RAPD markers
and less cumbersome than RFLP procedure (He and Prakash, 1997). According to
Powell et al. (1996) RAPDs yield more polymorphic markers than RFLPs but less
than AFLPs (Vilanova et al., 2001). AFLP technique might be more appropriate;
however, its higher cost does not give good reason for its use for cultivar
identification (Oliveira et al., 1999).
On the other hand, RAPDs became popular due to their efficiency,
simplicity, the relative ease to perform the assay and non-requirement of prior
information about DNA sequence (Khanuja et al., 1998).
RAPD markers are as efficient as AFLP markers (Ipek et al., 2003; Young
and Mark, 2004; Wen et al., 2004), ISSR markers (Martins et al., 2003) and SSR
markers (Zahuang et al., 2004) in the studies of genetic diversity (Sarkhosh et al.,
2006). They have been used in genetic diversity studies (Colombo et al., 2000;
Upadhyay et al., 2004; Sarkhosh et al., 2006, Cheng, 2007), phylogeny and
systematics (Sun et al., 1998), genetic linkage mapping (Cheung et al., 1997) and
gene tagging (Tiwari et al., 1998).
21
RAPDs, AFLPs and SSRs were compared in terms of their informative
ness and effectiveness in a study of genetic diversity and relationships among 32
cultivars of olive cultivated in Spain and Italy. All the three markers were found to
be highly efficient in discriminating the cultivars analyzed (Belaj et al., 2003a).
2.3.2.1 RAPD Markers
Williams et al. (1990) introduced the random amplified polymorphic DNA
(RAPD) method. This technique has efficiently been used for genome mapping in
plants (Staub et al. 1996; Mohan et al. 1997) and for the detection of plant disease
resistance genes (Michelmore et al. 1991; Martin et al.1991; Fazio et al. 1999).
Random amplified polymorphic DNA (RAPD) markers can be used for the
detection of DNA polymorphism without the requirement for predetermined
genetic data. Each product is obtained from a region of the genome that contains
two short fragments in inverted directions, on opposite strands that are
complimentary to the primer and adequately close together for the amplification to
work (Williams et al., 1990; Welsh and McClelland, 1990).
Quite a lot of studies have underscored the advantages of RAPD markers to
appraise the genetic diversity in different fruits. These markers have been used
dependably as molecular markers in cultivar characterization of peach (Chaparro et
al., 1994; Warburton and Bliss, 1996), plum (Ortiz et al., 1997), apple (Koller et
al., 1993; Yae and Ko, 1995), lemon (Deng et al., 1995) and grapes (Qu et al.,
1996) and were found highly appropriate to establish a molecular database for the
22
Psiadia species, and to discover the relationships and genetic homologies between
the species (Besse et al., 2003).
RAPD analysis has become extensively used to characterize and trace the
phylogeny of various plant and animal species (Dubouzet et al., 1997). The major
advantages of RAPD analysis over other techniques are its small sample DNA
requirement and the high frequency of polymorphic bands detected (Williams et
al., 1990).
RAPD analysis provides an uncomplicated and reliable method for
determining genomic variation. Because it is a comparatively simple technique to
apply, and the number of loci that can be detected is unlimited, RAPD analysis is
considered to be mor useful than RFLP's and other techniques (Lynch and Milligan
1994). This technique has further advantages over other marker systems of genetic
studies because it has a universal set of primers; no preliminary work such as filter
preparation, probe isolation or nucleotide sequencing is essential (Williams et al.
1990).
RAPD technique can detect sufficient polymorphism to make a distinction
among genotypes, even among the closely related cultivars. It can be helpful in the
identification of new cultivars as well as for the estimation of the genetic similarity
among different genotypes (Nicese et al., 1998). Genetic variations in the
populations are easily detectable using the RAPD technique with single-primer
amplifications (Vicario et al., 1995). According to Mulcahy et al. (1995) a single
primer may frequently be sufficient to discriminate all of the sampled varieties.
23
This can be used to characterize the DNA variation models within species
and among narrowly related taxa. RAPD markers have been extensively used for
the identification of genetic relationships among cultivars (Ba et al., 2004). The
use of RAPD markers proved effective in discriminating between even closely
related cultivars (Vilanova et al., 2001).
Since passport and pedigree data are time and again indefinite or
incomplete for many fruit and nut tree species (Warburton and Bliss, 1996),
RAPDs can be a valuable tool to appraise the degree of similarity of cultivars in
these woody species in order to choose the best parents to get new genetic
combination; it is especially important when a long generation time and as a result,
the long breeding process is involved in case of such perennial species (Nicese et
al., 1998).
RAPD markers are important not only for the characterization of the
germplasm but can also be used to assess the effects of selection over time and to
help in the development of cross breeding programmes since this process allows
the study of the genetic diversity of the existing germplasm (Nicese et al., 1998).
RAPD analysis offers a speedy, economical and stable way of generating a genetic
profile for the horticultural crops (Hormaza, 1999; Jordano and Godoy. 2000; Cai
et al., 2007).
This technique has the advantages of being rapidly employed, involving
very small amounts of genomic DNA and eliminating the need for blotting and
radioactive detection (Oliveira et al., 1999). The main advantages of RAPD are:
24
less laborious tests rapid scanning of the genome, greater number of loci per assay
and higher band-sharing and (Baranek et al., 2006).
A foremost advantage of RAPD markers over some other DNA based
markers is that they necessitate no prior sequence information, and no prior
knowledge regarding any particular gene in a target taxon (Palumbi, 1996).
Therefore, these markers can be used in the methodical study of new plant species
(Al-Humaid and Motawei, 2004).
RAPD markers can generate markers more quickly as compared to
restriction fragment length polymorphism (RFLP) markers, (Selbach and Cavalli-
Molina, 2000). They are comparatively more economical and faster to analyse,
when compared with the other marker systems. Moreover, their analysis does not
involve sophisticated laboratory equipments or skills (Creste et al., 2005).
The use of RAPD markers for the cultivar identification through DNA
profiling is the present method of choice in assessing the genetic variation
contained by germplasm collections (Bhutta et al., 2006). This technique is helpful
to develop genotype-specific banding patterns important for cultivar identification
(Gaafar and Saker, 2006).
2.3.2.2 Reproducibility of RAPD markers
Although RAPD has been extensively used for the construction of
phylogenetic relationships and it has the prospective to study phylogeny of plant
species, occasionally it has been controversial for its reproducibility (Kim et al.,
2005).
25
There are various factors which influence the reproducibility of RAPD
amplification profiles such as any difference in the process used for DNA isolation
(Korbin et al., 2000), concentration of primer or Taq-DNA polymerase, anealing
temperature, number of thermal cycles and concentration of MgCl2 (Bassam et al.,
1992; Kernodle et al., 1993), template quality and quantity, primer sequence and
the type of thermocycler (Hernendez et al., 1999).
In spite of the shortcoming caused by the lack of easy reproducibility,
RAPD markers can be of great importance as a fast process for taxonomic studies,
(Oliveira et al., 1999). A number of researchers have reported that majority of the
RAPD bands are reproducible if one take care while developing a standardized
procedure that is strictly followed in all the reactions (Hedrick, 1992; Gibbs et al.,
1994).
It is possible to avoid the poor reproducibility in RAPD analyses through
improvement in the laboratory techniques and the band scoring procedures
(Nybom, 2004). Improving the operator’s skill and standardizing the working
environment can also overcome this problem (Creste et al., 2005). However, the
use of a standardized RAPD protocol can guarantee the reproducibility of RAPD
patterns (Bhutta et al., 2006). In order to ensure high RAPD reproducibility, it is
important to optimize the PCR reaction (Gaafar and Saker, 2006).
Keeping in view that molecular markers other than RAPD technology are
very costly for assessment of minor fruit crops, RAPD emerges to be a good choice
for fingerprinting these species (Badenes et al., 2004) and looks to be more
appropriate for the studies of genetic diversity in fruit plants (Baranek et al., 2006).
26
Although newer methods like SSR and AFLP are ideal for their
informativeness, RAPD is still a better option for less sophisticated laboratories
because of its low cost, simplicity and lower infrastructure requirement (Upadhyay
et al., 2004). RAPD markers are even more appropriate than isozymes or
microsatellite markers for sorting out associations between different genera
(Guadagnuolo et al., 2001; Besse et al., 2003).
Morphologically very similar lines are found to be dissimilar at the
molecular level and such problems connected with taxonomical classification
highlight the need of complementary means for detection and characterization of
the genotypes. It is possible to find out a standard set of RAPD primers that can be
used to differentiate and characterize most of the familiar genotypes that thereby
serve as a valuable supplement to conventional agronomic and morphological data
for plant variety protection (Nandini and Chikkadevaiah, 2005).
There are numerous natural types in the fruit crops. A number of cultivars
are from seedling selections, various from crosses and some from mutation. It is
not easy to distinguish their parents. Randomly amplified polymorphic DNA
technique has been found very useful for ancestry analysis of fruit crops (Welsh
and McClelland, 1990; Williams et al., 1990).
2.3.2.3 Use of RAPD in loquat
An efficient sampling as well as accomplishment of germplasm resources
necessitates the exact identification of plant material. Molecular markers present an
efficient tool for cultivar fingerprinting, assessment of genetic similarity and
27
relationships. In a study, RAPD markers were tested as a means for loquat
germplasm management. Thirty-six primers were employed to screen 33 cultivars.
Twenty-three primers found polymorphic, they produced 29 polymorphic
amplification fragments, which were chosen as markers. Out of 33 accessions, 22
were identified by distinctive combination of RAPD markers. Four different
combinations were shared by two or more cultivars each (Vilanova et al., 2001).
RAPD-PCR has been used to find out the DNA polymorphism of 16 loquat
cultivars. Two of the used primers could amplify DNA fragments in these
cultivars. Nineteen DNA fragments were amplified in 16 cultivars by the two
primers. Three fragments were common while 16 were polymorphic or unique,
representing rich genetic diversity in the loquat cultivars. The results of DNA
fingerprinting demonstrated that 16 cultivars could be differentiated from each
other. The technique could be useful for the identification of bred cultivars and to
the marker-assisted breeding in loquat (Pan et al., 2002).
Sixty nine loquat (Eriobotrya japonica) accessions, including 27 varieties
from Japan, 23 from China, 4 from Greece, 7 from Israel, 3 from USA, 3 from
Mexico and the other 2 Eriobotrya species were analysed by random amplified
polymorphic DNA (RAPD). Out of the 60 primers, 28 primers yielded overall 135
fragments which were reproducibly amplified. Polymorphism was examined in
108 of the 135 bands. All varieties were efficiently distinguished by at least 1
band. It pointed out that RAPD analysis could be successfully applied to
distinguish loquat cultivars (Fukuda et al., 2002).
28
Germplasm collection from within Spain and the accessions introduced
from Italy, Japan and Portugal were studied by means of RAPD markers. Thirty
three highly reproducible markers were documented among 47 accessions. The
polymorphism obtained proved helpful to distinguish 39 accessions. Two groups
of accessions, which shared the same combination of RAPD markers, could not be
distinguished. According to the records and pomological characteristics, these
accessions corresponded to bud mutations (Badenes et al., 2003).
Eleven accessions of loquat including Eriobotrya japonica (cultivars
Moriowase, Jiefangzhong,Zaozhong No. 6, Baili, Wuqi, Luoyangqing, Hubeiliuer
and Yantangpipa), E. deflexa, E. prinoides and Guihouyesheng, wild loquat from
Guizhou Province, China, were analysed by random amplified polymorphic DNA
(RAPD) with 14 arbitrary decamer primers. A total of 130 DNA bands were
amplified, among which three bands were common. The degree of genetic
diversity was 97.9%. By UPGMA, 11 loquat genetic germplasms were separated
into 2 groups (red colour and white colour pulp group). Furthermore, the pulp
colour appeared to be a taxonomic indicator on DNA molecular level in loquat
(Chen et al., 2003).
The convenience of RAPD markers for genotyping a minor fruit species
such as loquat has been tested in order to evaluate their ability for recognizing
accessions and to make available a set of appropriate markers for use by a number
of scientists. Twenty nine polymorphic markers selected from an earlier study of
33 accessions were tested in 46 new accessions added to the collection. Using the
same standard conditions of PCR, only 20 markers out of 29 selected in the earlier
29
study gave consistent amplifications in the new set of plant material. Rest of them
required optimization of PCR conditions. This fact revealed that RAPD markers
were sensitive to the experimental environment, hence just a standard technique
did not ensure the reproducibility. To get through this problem, markers for plant
fingerprinting should be selected subsequent to the comparison across accession
sets. Only the markers which are reproducible with different sampling and
confirmed in several sets of accessions are appropriate for germplasm
fingerprinting. The markers obtained were sufficient for establishing origin and
relationships of cultivars, for recognizing synonyms and derived varieties from bud
sports. All bud sports were identical for all the selected RAPDs (Badenes et al.,
2004).
Recently, a new line of loquat ‘Chuannong No. 1’ has bee obtained by
breeding. For some reasons, its parentage history was unknown. The analysis of
this new line was carried out to discover its deriving cultivars by the comparison of
the genetic distance among this line and the three possible parents by using RAPD
markers. Five primers gave successful amplifications and polymorphism in the
loquat cultivars. Forty three bands were obtained, and sixteen out of them were
polymorphic. Cluster analysis gave an idea that ‘Longquan No. 5’ was likely the
parent of ‘Chuannong No. 1’ (Luo et al., 2007.
2.3.2.4 Use of RAPD in other fruit plants
The prospective use of the RAPD technique for characterization and
evaluation of genetic relationships was examined in nineteen walnut (Juglans regia
L.) genotypes used as parents or released as cultivars from the breeding program of
30
the University of California at Davis. Seventy two decamer primers were used,
most of which yielded scorable amplification patterns based on distinguishable
bands. A unique fingerprint was produced for every walnut genotype studied from
the results obtained. Cluster analysis divided the 19 genotypes into two main
groups whose dissimilarity was related to their heredity. Genotypes sharing
common parents were inclined to group together (Nicese et al., 1998).
RAPD markers were used to estimate the genetic resemblance among 35
mandarin accessions. One octamer and twenty two decamer primers generated 109
RAPDs, 45 of which were polymorphic. Jaccard coefficient was used to estimate
genetic similarity, and UPGMA to make the phenogram. The RAPDs obtained
were enough to produce some accession specific markers, and to divide these
accessions by clustering them into several groups. The genetic resemblance within
the mandarin group is high, and suggests the presence of narrow genetic base
among the cultivated mandarins. The genetic similarity of mandarins to other citrus
species was much lower (Filho et al., 1998).
Molecular characterization and phenetic relationships between some
cultivars of P. pyrifolia, and P. communis and genotypes of P. bourgaeana P.
cordata, and P. pyraster were examined through RAPD markers. Screening of
sixty decamer primers generated polymorphic patterns of Occidental and Oriental
pear genotypes. Twenty two selected primers created clear and reproducible
patterns and produced a total of 358 bands while 327 of them were polymorphic.
Out of 12 genotypes investigated, it was possible for 10 genotypes to locate
genotype specific RAPDs and fragment patterns which could be helpful for
31
cultivars identification. The patterns discriminated between genotypes and their
analysis set up a first approach to phenetic classification within the Pyrus genus
based on DNA markers, grouping the genotypes in accordance with their
geographic derivation. RAPD analysis of in vivo and in vitro material of seven
cultivars was also done which resulted in identical patterns for each genotype
(Oliveira et al., 1999).
DNA analysis was carried out on 18 Italian and exotic cultivars of Corylus
avellana that are imperative either for processed kernels or as table use, and one
cultivar of Corylus maxima with attractive characters. For the RAPD analysis,
oligonucleotide primers (10-mers) from Operon Technologies were tested. All of
the tested primers produced polymorphic bands having size between 500 and 3000
base pairs. Based on a set of 45 polymorphic markers, the genotypes were
compared and a phylogenetic tree was constructed by cluster analysis (Miaja et al.,
2001).
Genetic diversity studies were carried out in a set of 103 olive cultivars
using the RAPD technique from the World Germplasm Bank in Cordoba, Spain. A
total of 126 polymorphisms (6.0 polymorphic markers per primer) out of 135
reproducible products (6.4 fragments per primer) were obtained from the 21
primers used. The number of bands per primer varied from 4 to 11, while the
number of polymorphic bands ranged from 3 to 10, corresponding to 83% of the
amplification products. The pattern of genetic variation among olive cultivars from
three different Mediterranean zones (West, Centre and East) was analysed by
means of the analysis of molecular variance (AMOVA). Although most of the
32
genetic diversity was attributable to dissimilarity of cultivars within Mediterranean
zones (96.86%) significant φ-values among zones (φst = 0.031; p < 0.001)
proposed the existence of phenotypic differentiation. Most of the genetic diversity
was attributable to differences among genotypes within a country. It shows the
significance of the search of the distribution and amount of genetic diversity for a
better investigation of genetic resources of olive and the design of plant breeding
programmes. (Belaj et al., 2002).
Nineteen Albanian olive cultivars and two wild olives were studied to
determine their diversity level by using RAPD markers. A total of 76 polymorphic
bands were obtained using 16 primers (4.8 polymorphic markers per primer). The
number of bands per primer ranged from 4 to 10, whereas the number of
polymorphic bands ranged from 1 to 9, corresponding to 71% of the total
amplification products. A combination of 4 primers i.e. OPA-02, OPA-19, OPK-16
and OPP-19 could recognize all the accessions. The dendrogram, based on
Jaccard’s index, clustered the cultivars into three major groupings according to
their origin: the cultivars from the area of Berat, in South of Albania; cultivars
from the Centre and Centre North of Albania; and the cultivars from the Centre
and North West of Albania (Belaj et al., 2003b).
The RAPD technique was applied to study the genetic diversity and
relationships within the peach (Prunus persica L.) cultivars of the Czech Plant
Genetic Resources (PGR). The objective of the work was to construct a
dendrogram for assessing the genetic similarity and to separate the collection into
groups. 46 primers were applied to 6 peach cultivars from different places of the
33
country, having different the morphological characteristics like fruit shape and the
fruit colour. 12 primers were selected which gave polymorphic reproducible strong
and middle strong bands. They were afterward used for the RAPD reactions within
the entire peach collection. These RAPD primers differentiated 28 peach cultivars.
RAPD data were used to cluster the accessions analysed. Almonds and peach ×
almond hybrids were unmistakably divided in the frame of the complete
assortment. The combination kept up a correspondence to the botanical
classification, to the available pedigree information and to the cultivars description
(Raddova et al., 2003).
Indian coconut accessions were analyzed for the genetic diversity and
genetic relationship among them by using RAPD markers. DNA from 20
accessions of palm, 15 local and 5 exotic was amplified with 8 highly polymorphic
primers. These primers yielded 77 markers, with an average of 9.6 markers per
primer. Genetic diversity within accession ranged from 0.057 to 0.196. In general,
tall accessions were more heterozygous as they had higher degree of polymorphic
bands and genetic diversity. The amount of variation explained by within accession
and between accessions diversity was 0.58 and 0.42, respectively. In the same way
foreign accessions displayed more dissimilarity. Dwarfs from geographically
distant regions did not group separately (Upadhyay et al., 2004).
In a study, the phylogenetic relationships among Pyrus communis and
Pyrus pyrifolia were assessed using random amplified polymorphic DNA (RAPD)
and the conserved rDNA sequences. The pattern examined discriminated between
genotypes and their analysis established the approach to phenetic classification
34
within the Pyrus genus based on DNA markers, clustering the genotypes according
to their geographic origin. In RAPD analysis, UPGMA separated the cultivars into
two main groups; 19 P. pyrifolia cultivars and 6 P. communis cultivars (Kim et al.,
2005).
A simple and regular process for the analysis of tissue culture-derived
plants for somaclonal variations is a precondition for accurate monitoring of
quality control during mass micro propagation. In the same way, identification of
different varieties at molecular level is an essential element for efficient and
successful management of genetic resources. In Egypt, seven strawberry varieties
were screened using RAPD markers. Out of 20 RAPD primers tested, only four
were selected as producing polymorphic bands discriminating the investigated
cultivars. Based on those markers, the genetic distances between varieties were
found and their genetic relationships were estimated. It was revealed by the
phylogenetic tree that the cultivars showed close similarity within the cluster.
Although small morphological variations were evident in the leaves of some
clones, the developed RAPD profiles of different micro propagated clones were
typical to that of the mother plant (Gaafar and Saker, 2006).
RAPD technology was applied to study the genetic relationships between
36 Iranian, Russian, American and European almond cultivars and three wild
Amygdalus species. Thirty five 10-mer primers were used. All of them proved to
be polymorphic. Out of 734, 695 polymorphic bands were detected. Thirty five
polymorphic primers distinguished all the cultivars and species. Cluster analysis
grouped the cultivars studied according to their geographic origin or their pedigree
35
information. Iranian, American and European cultivars were clustered into three
separate groups (Kiani et al., 2006).
RAPD markers were used to find out the diversity level among 24 Iranian
pomegranate genotypes. One hundred decamer random primers were used for
PCR, among which 16 showed consistent polymorphic patterns. These primers
created 178 bands, out of which 102 were polymorphic. Cluster analysis of the
genotypes was carried out on the basis of data from polymorphic bands. The
highest and lowest relationships identified between genotypes were 0.89 and 0.29,
respectively. At 60% similarity, the genotypes were separated into four sub-
clusters. Cophenetic correlation coefficient between similarity matrix and
cophenetic matrix of dendrogram was relatively high (r = 0.9) demonstrating the
goodness of fit of the dendrogram. RAPD markers proved to be a valuable tool for
studying the genetic diversity of pomegranate (Sarkhosh et al., 2006).
The investigation of the prescreening data using 39 selected almond
accession and 80 RAPD primers illustrated that 42 primers produced true and
reproducible amplified products, which identified polymorphism among the
cultivars used. As well polymorphic primers were used for the investigation, a
relatively big number of polymorphic RAPD markers were distinguished by these
primers. While screening all the 39 cultivars and species, overall 729 bands were
revealed out of which 664 bands were polymorphic and generated 91.1%
polymorphism (Shiran et al., 2007).
Twenty six cherry genotypes (CC1 to CC26) from the Coruh Valley in
Turkey were assessed for genetic relationships by using RAPD markers, based on
36
56 decamer random primers. Seven out of them showed consistent
polymorphisms. These seven primers produced 80 markers, with 77 (96.25%)
exhibiting polymorphisms (Ercisli et al., 2007).
Differences among 8 cherry species and 2 inter specific progenies were
analyzed through the use of RAPD technique. Forty eight random oligonucleotide
primers were screened for PCR amplification to generate polymorphisms. The
diversity analysis was conducted using two distance-matrix methods. The
dendrogram was constructed to know the relationships among them. The results
revealed that in total, there were 840 amplified loci; 23 sweet cherry and 4 sour
cherry cultivars grouped together with 569 and 247 polymorphic loci respectively,
accounting for 67.74% and 29.40% of the total dissimilarity. Prunus fruticosa,
Prunus tomentosa and Prunus humilis produced a monophyletic group. A
relationship between Prunus pseudocerasus L. and Colt, which made a further
closely related group, was detected while Prunus cerasus, Prunus avium and other
cherry species were more different. The range of genetic remoteness was from
0.0623 to 0.2719 among the Prunus species, which were discrete by inheritance.
The topology of the tree was by and large in agreement with the taxonomic
categorization. The results pointed out that with the exception of ‘Hongdeng’
variety, there were one or more cultivar specific RAPD markers in the species and
cultivars studied. By the use of these specific markers, cherry species and varieties
may possibly be identified, So there is the potential to select excellent characters of
hybrids at an earlier phase (Cai et al., 2007).
37
DNA of 180 accessions in 10 demes in Prunus persica were amplified with
22, 10-base primers chosen from 200 random primers using RAPD technology.
One hundred and eighty loci were detected. With statistical analyses of the data,
genetic variation of the demes was expressed. Genetic diversity among and within
groups were 11.9 and 88.1% respectively. Through analyses of genetic variation
and genetic arrangement, the results could provide molecular biological facts for
conservation and utilization of P. persica germplasm (Cheng, 2007).
In an investigation, forty two pecan cultivars could be distinguished with
one or more promers on the basis of RAPD fingerprints. Seven cultivars: ‘Colby’,
‘Giles’, ‘Money Maker’, Evers’. ‘Elliot’, ‘Summer’ and ‘Wichita’ could be
distinguished through the absence of presence of a single RAPD band.
Identification of remaining cultivars required at least two bands to be scored
(Conner, 2008).
2.3.2.5 Molecular markers – supplement to conventional research
Morphological markers cannot be substituted by any of the molecular
techniques for the purpose of characterization. However, the results of molecular
or biochemical studies should be considered as complementary to morphological
characterization (Karp et al., 1997). Characterization of accessions in a germplasm
by applying molecular markers or agronomic characteristics is a general practice,
but simultaneous use of both in the research studies is not as much frequent
(Bramardi et al., 2005).
38
Genetic diversity among Iranian pomegranate genotypes was studied on the
basis of fruit characteristics with the simultaneous use of RAPD markers.
Grouping was made in accordance with the fruit characteristics. Among 113
random primers tested, 27 revealed good quality amplification and polymorphism,
and a total of 158 RAPD markers were formed. It was revealed by the study that
information based on fruit characteristics alone is not enough for the assessment of
genetic diversity (Zamani et al., 2007).
Recently, a new white flesh loquat cultivar, ‘Ninghaibai’ has been
characterized with new molecular marker procedures combined with the method of
conventional botany description. This combined use of both the procedures was
helpful to distinguish the ‘Ninghaibai’ cultivar from other eleven cultivars of
loquat (Feng et al., 2007).
2.4 Evolution of varieties
History shows that loquat species was grown as an ornamental tree having
small sized fruits and was introduce to Europe as ornamental (Polat and Caliskan,
2007; Soler et al., 2007). Through thousands of years of selection a lot of cultivars
have been selected with excellent quality and large sized fruit (Janick, 2007). Over
and over again, natural hybridization resulted in the variation in the seedlings and
selection of new varieties (Daito, 1995). Several studies have described
characteristics of promising loquat cultivars obtained by selection from natural
variation or breeding in China (Huang et al., 1993), Brazil (Athayde et al., 1992),
India (Singh and Lal, 1989) and Italy (Monastra and Insero, 1992; Baratta et al.,
39
1995). New cultivars of loquat obtained by radiation breeding have been reported
in China (Badenes et al., 2000).
Presently, a number of breeding programs are in progress and new cultivars
have been developed that show adaptation to different localities (Janick, 2007).
Japan has great contribution towards the development of loquat. Japanese
horticulturists selected two outstanding cultivars, ‘Tanaka’, and ‘Mogi’ from the
progeny of seedling introduced from China (Lin et al., 2007). Loquat is becoming
an important industry in China as well as Spain, Japan, India, Pakistan and Turkey
(Janick, 2007).
2.5 Status of Germplasm resources of loquat
According to Yu (1979), the genus Eriobotrya consists of 16 species. Only
E. japonica is grown for its fruit. Other species of the genus are raised as
rootstocks or as ornamentals (Morton, 1987; McConnell, 1989; Lin, 2007). There
are a lot of cultivars and selections of E. japonica in different provinces of China.
For example, there are 83 cultivars in Zhejiang, 57 in Jiangsu, 78 in Fujian, 31 in
Anhui, 18 in Guangdong and 9 in Sichuan (Zhang et al., 1990). Forty cultivars
were registered in Japan (Fujisaki, 1994). The biggest collection of germplasm,
having more than 250 cultivars, is situated in Fuzhou, China, while less than 50
cultivars are commonly cultivated in the country. Spain and Japan have germplasm
banks with 100 cultivars and 60 cultivars respectively. Italy has a collection of 16
cultivars (Lin, 2007). A white flesh loquat garden and a hybrid strains
garden have also been established in China (Lin, 1990; Zheng, 2007).
40
There are two main types of loquat: the ‘Chinese type’ loquat, which is
characterized by large sized fruits that are pear shaped with yellow flesh colour and
the ‘Japanese type’ loquat, having small sized fruits that are round shaped with
white or pale yellow flesh colour (CTIFL, 1988)., A large number of varieties have
emerged from these types in the different loquat growing countries (Badenes et al.,
2000). Cultivation of the species has resulted in the evolution of a large number of
new cultivars, due to different selection pressures applied by the growers. With the
increase in its cultivation, the number of new cultivars also increased (Vilanova et
al., 2001).
2.6 A brief description of some loquat cultivars
Loquat cultivars are going to be described since long on the basis of
morphology of their fruit, tree, leaf or flowers. A brief description of a few loquat
cultivars based on morpho-physical traits as given by different authors is as under:
‘Guangyu’ is a leading variety, grown on commercial scale and area under
this variety is being extended. Its fruits are large weighing 43.5 - 61.5 g.
Sometimes, its fruit weight reaches 75 g. They have good eating quality. The flesh
is excellent, tender and juicy, with a rich flavour and soluble solids content of
13.4%. The percentage of edible portion reaches 71% and its shelf life is good (Xu
et al., 2000).
‘Hongdenglong’ is a chance seedling which was obtained in 1987. It is a
promising late loquat variety that matures in mid or late June and has a very good
eating quality. Fruit size is large, with average weight of 63.1 g but it may reach
41
over 100 g. The average number of seeds per fruit is 4.10. Colour of the skin is
orange red, with a few spots. Flesh colour is orange red, fine, juicy and tender,
with a soluble solids content of 13.5% and an enjoyable acid sweet aroma (Jiang et
al., 2001).
‘Yangmeizhou 4’ is a chance seedling, which was discovered in 1980. It is
a hardy and high quality cultivar of loquat. Its fruit matures in the last week of
May. Fruit size is medium to large and weighing approximately 32.2 to 54.5 g.
Fruit shape is round or ovate and skin colour is orange yellow. Flesh is also orange
yellow and juicy with a soluble solids content of 11.7-13.8% and very pleasant
flavour. The percentage of edible portion of the fruit reaches 67.2 % and the skin
can be peeled quite easily. Number of seeds per fruit is 2.9 on average. The trees of
‘Yangmeizhou 4’ are fruitful and can tolerate low temperature of -12.8°C in the
winter (Wu, 2001).
‘Donghuzao’ is an early variety of loquat, which was obtained as a chance
seedling in China.. Trees are precocious and resistant to leaf spot, which give an
average yield of 22.5 kg per plant at an age of four years. Fruit matures from late
March to early April. Fruit is large sized and round shaped, having an average
weight of 59.2 g, and may reach upto 110 g. Skin colour is orange red with a few
rust spots on it. Flesh is also orange red in colour, having a soluble solids content
of 9.0 to 11.0 % and a pure sweet flavour. Fruit is of very good eating quality and
no cracking was observed in it (Zhao et al., 2001).
‘Taicheng 4’, is also an early variety, which was selected as a chance
seedling. It has an average fruit weight of 42.5 g, and only 1.34 seeds per fruit.
42
‘Jiefangzhong’ is the most important variety grown in Fujian province of China. It
produces the largest fruits of any loquat variety, fruit weight reaching 172.0 g
(Zheng, 2001).
‘Puxinben’ is another chance seedling which was selected in China in 1993.
Its excellent characteristics include large sized fruits of good quality and high
production. Fruit matures from early to mid May, weighing 58.1 to 74.1 g with a
attractive orange yellow skin. It is easy in peeling. The flesh is tender, fine and
juicy with soluble solids content of 10.2 to 12.6% and an enjoyable acid sweet
flavour. Two to three seeds are present in each fruit. Fruit is of very good eating
quality with an edible rate of 68.8 to 72.6 % (Peng et al., 2002).
‘Donghuzao’ is a new loquat variety resulting from a seedling discovered in
1986. It is a very promising early variety, which matures from late March to early
April in China. Fruits and are very large, having weight of 50 to 60 g on average,
but reaching 110 g. Colour of fruit skin is orange red, which is easy to peel. Flesh
fine, juicy and tender with an orange red colour. Fruits have a soluble solids
content of 10.6 to 11.2 % and the fruit juice contains an ascorbic acid content of
56.1 to 75.7 mg/L. The trees are precocious and productive, giving a yield of 24.8
kg of fruit per tree at the age of 5 years (Zhao et al., 2003).
Ninghai, in China, has the most favorable climate for cultivation of loquat,
with an annual precipitation of 1200 to 1500 mm. An average annual temperature
is 15°C, and minimum temperature in the cold months may reach -5°C. The white-
fleshed cultivar ‘Luoyangqing’ and red-fleshed cultivar ‘Dahongpao’ are the 2
major cultivars grown. The new cultivar, Ninghaibai, is also extensively planted.
43
'Ninghai Bai', was developed by an organized selection method from seedling. It is
characterized by large fruit size and white flesh colour. Its fruit shape is round or
long round. Weight of a single fruit is 40 to 65 g; the heaviest reaching 86 g. Fruit
is of good eating quality having an edible percentage of 73.6 %. The fruit colour is
light yellow white, the skin is thin, and the flesh is fine and sweet-smelling. The
total soluble solids content is 13 to 16% on average, the highest value reaching
19.2 %. The fruit matures during the end of May. The cultivar is high yielding and
resistant to freezing (Feng, 2003; Feng et al., 2004).
The fruit characteristics of 10 loquat cultivars were observed and compared
in a study conducted in China. Results showed the best one was ‘Guanyu’ cultivar,
which was followed by ‘Yangshi 1’. ‘Guanyu’ has a large sized fruit, weighing
45.2 g on average, but reaching 70 g. It has a soluble solids content of 12.6 % and
an edible percentage of 70 %. ‘Yangshi 1’ also has large fruit size, weighing 40 g
on average, which may reach 80 g, with a soluble solids content of 14.8 % and an
edible percentage of 65 % (Lou et al., 2004).
‘Dawuxing’ is another promising loquat cultivar developed in China. Fruit
matures during mid May, having a round or elongated round shape, with a yellow,
golden yellow or orange yellow skin colour. Weight of fruit is 43.3 to 57.0 g, with
a soluble solids content of 8.0 to 9.8 % and an edible portion of 70 to 80 %.
Number of seeds per fruit are 1 to 3 (Zhou et al., 2004).
‘Zaozhong 6’ loquat cultivar was developed by the Fujian Fruit Research
Institute, China. It was derived from a cross between ‘Jiefangzhong’ and the
Japanese variety ‘Shenweizaosheng’. It is an extra early loquat cultivar which is
44
commercially cultivated in loquat growing areas. It matures 15 to 20 days earlier
than the existing early cultivars. Its fruits are large sized, having an average weight
of 52 g, but reaching 116 g. It has a soluble solids content of 12 % and an edible
proportion of 70 % with an excellent eating quality. Application of fertilizers,
manures and fruit thinning is also essential for high quality and production (Zheng,
2001; He and Zhang, 2005).
Trials carried out in Meizhou region proved that ‘Maomu’ loquat cultivar
performed well under the local conditions. The trees are hardy. Average fruit
weight is 21.7 g, with 1 to 2 seeds per fruit and a soluble solids content of 12 %.
Eating quality is excellent (Luo, 2005).
Loquat cultivar, ‘Piera’ is a spontaneous bud mutation of ‘Algerie’, which
flowers, and gives fruit repeatedly during the year. It has greater fruit weight (59.3
g) than that of ‘Algerie’ (57.2 g). Up to 13 flowering flushes and subsequent fruit
set, included mainly in 3 groups, have been identified in this variety. ‘Piera’ gives
flowers from flower buds and mixed buds indistinctly. Fruit growth and
development occurs in three distinct periods. Only fruit developed during summer
becomes inferior in size having a weight of 9.8 g and is not commercially
acceptable. Accordingly, three harvest dates have been identified throughout the
year (Reig and Agusti, 2007).
“Luoyangqing’ cultivar is differentiated by the loose panicles, almost 12
cm long and 10 cm wide, having 58 to 134 flowers per panicles. Fruit weight
ranges from 31.60 to 35.20 g, having skin with orange red colour at maturity. Skin
is tough and thick but easily pealed off. The flesh is thick and juicy, orange red in
45
colour. The seed 2 to 4 per fruit are around 2 g. Edible portion is about 66.5 to 67
%. Fruit harvest begins in early May and ends in late May (Yang et al., 2007).
Guangxi, is one of the regions of China, which are rich in wild resources of
loquat germplasm. Huang et al. (2007) described the newly recorded cultivars of
loquat over there. One of them is Kumquat loquat, which is a clone of the wild
loquat. It gives flowers in August / September and matures during March / April.
Colour of fruit skin as well as pulp is orange. Weight of fruit is about 18 g with 3
to 4 seeds. Taste is sweet and slightly sour, while TSS is 13 %. It is can resist cold
and diseases. There is another cultivar named as Sour loquat, having comparatively
small sized fruit. It is a seedling that is slightly later in flowering and fruit bearing.
Its flowering starts in September, while ripening time is March / May. Average
fruit weight is 10.60 g with 3.30 seeds per fruit. Skin as well as pulp colour is
orange. TSS ranges from 8.00 to 10.00 %. Although it is sour in taste, it has a
strong resistance against cold and diseases.
At Dongling, China, a cultivar has been recorded with the name of ‘White
loquat with single seed’. It is also a seedling selection. It gives flowers in
September / October and matures in March / April. Fruit weight is 31 g with 1 or 2
seeds per fruit. Skin as well as pulp colour is yellowish white, while TSS is 14. It
has a sweet juicy flavour. It has more resistance against cold and diseases than
other cultivars (Huang et al., 2007).
Recently, a study was carried out in Turkey to select the loquat genotypes
with better quality. Selection of thirteen ‘Types’ was done on the basis of fruit
quality characteristics. Fruit weight of these types ranged from 20.5 g to 39.2 g,
46
fruit width and length ranged from 29.8 to 40.7mm and from 32.6 to 41.6 mm
respectively. Fruit width / length index of genotypes ranged from 0.89 to 1.06.
Average seed number per fruit and seed weight per fruit ranged from 2.0 to 5.3 and
from 2.9 to 7.4 g respectively. The flesh / seed ratio ranged from 3.88 to 5.10 in the
loquat types. When all the characteristics were taken into account, ‘Type 4’ and
‘Type 7’ were found better among these genotypes because of their fruit size,
while ‘Type 5’ was preferable for having low seed number and good flesh / seed
ratio (Polat, 2007).
He et al. (2007) investigated the loquat resources in Guandong, China.
They described four loquat cultivars; one grafted tree and three seedlings. ‘Taishan
Zhong’ is an early cultivar that ripens in the mid or late January, which is two
months earlier than local cultivars and one month earlier than ‘Zaozhong No.6’. Its
flesh is white in colour, and very high in sugar with an excellent flavour. It gives
good yield. On the other hand, fruit is small in size (12.8 g) with 1 to 3 seeds per
fruit. Length and width of leaf are 16.1 cm and 14.3 cm respectively TSS is 16.85
%. ‘Mojia No.1’ is a mid late cultivar, with orange colour and sweet flesh,
agreeable to the taste of Cantonese. Its skin is thick and tough with good shelf life.
Fruit weight is 53.2 g on average with 1 to 3 seeds per fruit. Length and width of
leaf are 19.8 cm and 5.2 cm respectively. TSS is 9.07 %. The tree canopy is loose
with long branches, which is inconvenient for cultural practices and fruit pocking.
‘Hanwuzhong’, another cultivar of loquat, has an average quality, but its growth is
vigorous and can establish a productive crown quickly. Fruit weighs 30.97 g with 2
to 4 seeds per fruit. Flesh is yellow while TSS is 9.92 %. ‘Qingbian’ is a grafted
tree (mother plant died) and has good quality and sells on higher prices than other
47
cultivars, but its production is low. Skin colour is yellowish while colour of flesh is
white. Weight of fruit is 28.7 g with 1 to 3 seeds per fruit. It has a TSS of 10.55 %.
The germplasm collection in Spain includes 90 accessions that are being
studied by pomological characteristics and molecular markers. Llacer et al. (2003)
have described the main characteristics of some of the loquat cultivars included in
the germplasm collection, which are as under:
‘Magdal’ is a medium vigorous cultivar.. The flower cluster is conical in
shape with a high number of flowers (178) per cluster. The fruit is long obovate
shaped, the skin and the flesh are both yellow orange in colour. The average fruit
weight is 45.50 g while fruit diameter is 36.60 mm. The seed shape is elliptical
with an average weight of 7.8 g. There are about 3.7 seeds per fruit.
‘Cardona’ is a medium vigorous culivar with upright tree habit. The flower
cluster is conical in shape having 168 flowers / cluster on an average. Shape of
fruit is round elliptic, the skin and the flesh are both yellow orange, the average
fruit weight is 45.4 g and fruit diameter is 41 mm. The seed shape is elliptic with
an average weight of 6.3 g. There are about 2.7 seeds per fruit.
‘Italiano-1’ is a very vigorous loquat cultivar. Tree habit is semi-upright.
The flower panicle is medium in size, having 160 flowers on an average. The fruit
shape is oblate, the skin and the flesh are both orange in colour, average fruit
weight is 51.40 g, while fruit diameter is 45.20 mm. The seed shape is ovate with
an average weight of 6.50 g. There are about 3.90 seeds per fruit.
48
‘Algerie’ is a vigorous cultivar of loquat. Tree habit is upright. Full bloom
occurs during the first week of November. The flower cluster is conical in shape
with a high number of flowers per panicle (an average of 200 flowers / panicle).
Fruit ripening time is during the first week of May (May 3rd, average). Shape of
fruit is round elliptic, the skin and the flesh are both yellow orange in colour, the
average fruit weight is 65.00 g and fruit diameter is 50.00 mm. It is easy to peel
from stalk end and has a good flavor. The seed shape is elliptic with an average
weight of 7.30 g. Number of seeds per fruit is about 2.3.
‘Golden Nugget’ is a very vigorous cultivar. Tree habit is semi spreading.
The flower cluster is intermediate in size having a high number of flowers per
cluster (an average of 189 flowers / panicle), white in color. The fruit is abovate,
the skin and the flesh are both yellow orange in colour, the average weight of fruit
is 54.60 g and fruit diameter is 45.30 mm. Flavor is relatively poor. The seed shape
is round with an average weight of 8.10 g. There are about 3.20 seeds per fruit.
‘Buenet’ is a medium vigorous cultivar. Tree habit is upright. The flower
cluster is conic shaped, with a high number of flowers per cluster (an average of
227 flowers / panicle). The fruit is round elliptic in shape, the skin and the flesh are
both orange in colour, the average fruit weight is 58.20 g and fruit diameter is
43.10 mm. The seed shape is elliptic with an average weight of 7.20 g, while there
are about 2.50 seeds per fruit.
‘Crisanto Amadeo’ is a vigorous loquat cultivar. Tree habit is upright. The
flower cluster is conical in shape, with a high number of flowers per panicle (an
average of 210 flowers / panicle). The fruit shape is round elliptic, the skin and the
49
flesh are both yellow orange in colour, very good in falvour. The average weight of
fruit is 68.70 g and diameter is 50.60 mm. The seed shape is round with an average
weight of 7.90 g, there are about 3.60 seeds per fruit.
‘Saval-2’ is a medium- vigorous cultivar of loquat. Tree habit is upright.
The flower cluster has a conical shape, with a high number of flowers per panicle
(an average of 273 flowers / cluster). The fruit is round, the skin and the flesh are
both yellow-orange in colour, the average fruit weight is 53.70 g while the
diameter is 43.90 mm. The seed shape is elliptic with an average weight of 8.60 g.
Average number of seeds per fruit is about 3.90.
‘Peluches’ is a very vigorous cultivar. Tree has a spreading habit. The
flower cluster is conical, with a high number of flowers per panicle (an average of
229 flowers / panicle). The fruit is long obovate in shape, the skin and the flesh are
both yellow-orange in colour, the average fruit weight is 95.0 g and diameter is
51.2 mm having about 3.7 seeds per fruit. The seed shape is elliptic with an
average weight of 11.2 g.
‘Tanaka’ is a medium-vigorous cultivar. Tree habit is upright. The shape of
panicle is conical, with a medium number of flowers per panicle (an average of
167 flowers / panicle). The fruit is abovate in shape, the skin and the flesh are both
yellow-orange in colour, the average fruit weight is 60.60 g and diameter is 48.50
mm. There are about 3.7 seeds per fruit. The seed shape is elliptic with an average
weight of 9.50 g.
50
2.7 Trends in loquat research
2.7.1 China
Loquat was an underutilized fruit species before the establishment of
modern China as breeding work was nonexistent. The rapid increase (from 2000 ha
in 1970 to 26,000 ha in 1995 and to 120,000 ha with an output of 460,000 tonnes
in 2005) was the outcome of new technology including genetic improvement that
resulted in evolution of new cultivars such as ‘Dawuxing’ and ‘Zaozhong No.6’,
improved planting techniques, the extensive use of grafting to seedling rootstocks,
flower and fruit thinning, and fruit bagging (Lin et al., 2007). After the
commencement of loquat breeding programmes, loquat resources were studied by
many research scientists all over the country. The rich germplasm resources were
evaluated and germplasm exploitation and utilization was initiated. Breeding of
loquat has been carried out regularly by means of introduction, selection and
crossbreeding. Many new cultivars were released and are currently cultivated in
large production areas (Huang et al., 1993; Zheng, 2001; Peng et al., 2002; Feng et
al., 2004; Zheng, 2007).
China covers a lot of aspects of loquat research. A lot of work has been
conducted including collection, investigation and preservation of germplasm
resources. From the late 1970s to the early 1980s, the National loquat germplsm
garden was established in Fuzhou, Fujian province (Lin. 1990; Wu, 2001; Zhao et
al., 2003; Feng et al., 2004; Zheng, 2007). In addition to collection, investigation
and preservation of loquat germplasm, work with reference to identification and
classification of loquat is also going on. Cytology and molecular biology
51
techniques have extensively been used in loquat research. PCR technology has
been effectively used to identify the center of origin and the evolution of species
and to classify and identify loquat germplasm resources (Pan et al., 2002; Chen et
al., 2003; Meng et al., 2003; Feng et al., 2007; Luo et al., 2007). Attention is also
being paid to introduction from other countries as well as from the old producing
areas of China (Zhao et al., 2003; Feng et al., 2004; Zheng, 2007).
Another feature of research in China is to search out the seed less loquat.
Unluckily, loquat has many large seeds, which are an undesirable character. It has
been a dream of several loquat scientists to get seedless loquat. Previous
techniques to obtain seedless clones include radiation (Li and Zhuo, 1991), use of
growth regulators (Gu, 1990; Sheng and Wu, 1998), endosperm culture (Chen,
1983) and via tetrajploid x diploid crosses, but these techniques so far have not
been successful. Now this problem has been overcome by mass screening of seed
to identify natural triploids that occur at a very small frequency (Liang et al.,
2007). A number of natural triploids have been found from 21 cultivars of loquat
during 1997 to 2005. These triploids are now at bearing stage and are seedless with
high quality (Guo et al., 2007).
2.7.2 Spain
Although, loquat reached in Europe as an ornamental tree with small sized
fruits (Soler et al., 2007; Polat and Caliskan, 2007), it is now an important minor
fruit in Spain. Spain is the second loquat producer and the first exporting country
in the world (Caballero and Fernandez, 2003; Soler et al., 2007; Canete et al.,
2007; Hueso et al., 2007). The National Institute of Agricultural Research
52
established a loquat collection in 1993. Afterward, a European project funded the
continuation of the European germplasm collection at Valencia. Since then more
than 130 accessions have been introduced. The collection is being extended by
surveys and exchange of accessions (Gisbert et al., 2007a).
Research is being carried out on various aspects including characterization,
breeding and management practices. Many loquat accessions have been
characterized on the basis of morphological characters (Martinez-Calvo et al.,
2000), RAPD markers (Vilanova et al., 2001; Badenes et al., 2003; Badenes et al.,
2004) and SSR markers (Soriano et al., 2005; Gisbert et al., 2007b). Work on
propagation techniques (Castro et al., 2007) fruit size improvement (Agusti et al.,
2007) and earliness of flowering (Cuevas et al., 2007) is also going on. Fruit size
and earliness are the most important factors in the commercialization of loquat.
Commercial size is usually achieved by means of heavy thinning either at bloom
(removing the upper two-thirds of the panicle), or in January (leaving 4 to 5 fruits
per cluster). Due to the high cost of labor in Spain a chemical alternative has been
developed using naphthalene acetic acid (NAA) and its derivates (Agusti et al.,
2000; Cuevas et al., 2004). Earliness can also be achieved by protected cultivation
(Lopez-Galvez et al., 1990) and by means of regulated deficit irrigation (Cuevas et
al., 2007). Dwarfing rootstocks such as ‘Quince C’ are under evaluation (Hueso et
al., 2007) to reduce spacing and management cost and therefore to increase
profitability (Lin et al., 2007).
53
2.7.3 Turkey
In Turkey, a quick increase has been observed after 1980. The total
production in 1980 was only 3000 tonnes, which increased to 9000 tonnes by 1990
and 12000 tonnes by 2003 (Polat and Caliskan, 2007; Karadeniz and Senyurt,
2007). Investigations on loquat are centered at Mustafa Kamal University in Hatay,
Cukurova University in Adana and Citrus Research Institute in Antalia.
Investigations are in progress on cultivar trials, resistance of cultivars to scab,
resistance to winter cold and spring frost, various propagation techniques,
germination of seeds, fruit thinning, parthenocarpy, high density and protected
cultivation (Demir, 1989; Polat and Kaska, 1991; Paydas et al., 1992; Yalcin and
Paydas, 1995; Polat, 1999; Polat and Caliskan, 2007; Polat, 2007).
54
Chapter 3
MATERIALS AND METHODS
3.1 Survey of the loquat growing areas
Loquat (Eriobotrya japonica Lindl.) growing region spreads from the
northern part of the Punjab province (generally known as Pothohar Plateau) to
Mardan district of the NWFP. Survey was conducted in the loquat growing areas
of Punjab and NWFP. Loquat growers, contractors and marketers were interviewed
to get the first hand knowledge regarding the production, marketing, sizeable
orchards and different genotypes of this fruit available at various places.
3.1.1 Selection of sites
Among various sites surveyed, nine sites were selected on the basis of their
suitability with respect to the availability of diversity in genotypes and the
cooperative attitudes on the part of the growers. Plants of different loquat
genotypes with distinct characters available at all these sites were selected and
permanently tagged. The selected sites include Kalar Kahar, Choa Saiden Shah,
Chhattar, Tret, Hasan Abdal, Wah, Hari Pur, Mardan and Takht Bhai
(Fig 1). Geographical location of these sites is given in the Table 1.
55
Fig.1 Map of Pakistan showing main loquat growing area (highlighted with yellow colour) and the sites selected for the study (red spots)
56
Table 1 Geographical location of the selected loquat sites
Location District Province Longitude Latitude Kalar Kahar Chakwal Punjab 72° 42' 00" E 32° 46' 60" N Choa Saiden Shah Chakwal Punjab 72° 59' 00'' E 32° 43' 00'' N Chhattar Islamabad Punjab 73° 10' 00'' E 33° 10' 00'' N Tret Rawalpindi Punjab 73° 17' 00'' E 33° 50' 00'' N Hasan Abdal Rawalpindi Punjab 72° 41' 08'' E 33° 49' 19" N Wah Rawalpindi Punjab 72° 45' 00" E 33° 46' 12" N Hari Pur Hari Pur NWFP 72° 55' 12" E 34° 01' 12" N Mardan Mardan NWFP 72° 02' 00'' E 34° 12' 00'' N Takht Bhai Mardan NWFP 71° 55' 39'' E 34° 16' 48'' N
57
3.1.2 Selection and tagging of plants
After repeated visits and observations, three plants of each genotype
with distinct characters available at each site were selected for the detailed
study regarding morpho-physical characteristics and RAPD analysis. Care
was taken to select the bearing plants with apparent good health at each
site with almost similar size and stem girth. Codes were assigned to each
genotype and permanent tags were attached with all the selected plants.
The brief view of the selected locations and the genotypes included in the
study is given below:
3.1.2.1 Kalar Kahar
Kalar Kahar is an historical place of district Chakwal in the Punjab
province. It is situated on the main Motorway of the country (M2) having
interchange link and Service area. It is quite famous for loquat production. Here,
the famous ‘Takht e Babari’ (Throne of Babar) is situated in the form of a rock
amid the loquat orchards, where the founder of Mughal Empire, Zaheer-ud-Din
Babar used to address his armies during early 16th century. He had established the
Royal Garden at Kalar Kahar. Presently, a lot of small and big orchards of loquat
are scattered in this area. Five genotypes were identified in a private orchard over
here. Different codes were assigned to all the five genotypes, which are; KK1,
KK2, KK3, KK4 and KK5.
58
3.1.2.2 Choa Saiden Shah
Choa Saiden Shah is situated in the south east of Kalar Kahar, district
Chakwal in the Punjab province. It is an old town and centre of commercial
activities including loquat fruit production and marketing. Coal and salt mines are
abundantly found over here. Three genotypes were identified in the district
government orchard at Choa Saiden Shah. Different codes were assigned to these
three genotypes, which are; CS1, CS2 and CS3.
3.1.2.3 Chhattar
Chhattar is situated at a short distance from Islamabad, the capital city. It
has been famous for loquat orchards for a long time, but presently most of the
orchards have been eliminated due to urbanization / constructions during the last
four decades after the time when the capital of the country shifted from Karachi to
Islamabad. At Chhattar, three genotypes of loquat were identified in the orchard of
a private grower. Different codes were assigned to these three genotypes, which
are; CH1, CH2 and CH3.
3.1.2.4 Tret
Tret is a mountainous place on the way from Islamabad to Murree, having
an elevation of 3420 feet. Loquat cultivation is quite successful in some pockets of
this area. Here five genotypes were identified at the experimental orchard of the
provincial government. Different codes were assigned to these five genotypes,
which are; TR1, TR2, TR3, TR4 and TR5.
59
3.1.2.5 Hasan Abdal and Wah
Hasan Abdal is an historical town in northern Punjab. It is located where
the Grand Trunk Road meets the Karakoram Highway near North West Frontier
Province. On the nearby hill, there is a meditation chamber related to a 15th century
Muslim Saint, Baba Wali Qandhari, popularly known as Baba Hasan Abdal.
Adjacent to Hasan Abdal, there is another town, Wah, which is famous for its
productive orchards. A number of small and large loquat orchards are scattered in
the areas of Hasan Abdal and Wah. Five genotypes were identified over here.
Different codes were assigned to these five genotypes, which are; HW1, HW2,
HW3, HW4 and HW5.
3.1.2.6 Hari Pur
Hari Pur is a city in the North West Frontier Province of Pakistan, 65 km
north of Islamabad, in a hilly plain area. This place is famous for high yielding
loquat orchards of good quality. Three loquat genotypes were identified at Haripur
in the orchard of a private grower and fruit contractor. Different codes were
assigned to these three genotypes, which are; HP1, HP2 and HP3.
3.1.2.7 Mardan
Mardan is an important loquat growing district of North West Frontier
Province of Pakistan. Three genotypes were identified in a private orchard near the
district head quarter. Different codes were assigned to these three genotypes, which
are; MN1, MN2 and MN3.
60
3.1.2.8 Takht Bhai
Takht Bhai is a place in Mardan district. Near Takht Bhai, there is a loquat
orchard belonging to Mr. Ikramullah Khan, a progressive grower of NWFP. He has
a good collection of loquat genotypes over there which he maintained during the
last five decades. Fifteen loquat genotypes were identified at Takht Bhai. Different
codes were assigned to these genotypes, which are; TB1, TB2, TB3, TB4, TB5,
TB6, TB7, TB8, TB9, TB10, TB11, TB12, TB13, TB14 and TB15.
3.2 Evaluation of loquat genotypes on the basis of morpho-physical
characteristics
Most of the traits studied were those described by UPOV (1995). To reduce
the environmental effects, data from two crop years (2005-06 and 2006-07) were
used. Following morpho-physical characteristics of the trees, leaves,
inflorescence, fruits and seeds were studied:
3.2.1 Morphological characteristics
3.2.1.1 Tree habit
Tree habit of the genotypes was noted as upright, semi upright or
spreading.
3.2.1.2 Shape of leaf tip
Shape of leaf tip was observed for the different genotypes as
sharp acute and blunt acute.
61
3.2.1.3 Shape of panicle
Shapes of panicle for different genotypes included conical,
truncate conical and cylindrical.
3.2.1.4 Fruit morphology
Parameters for this purpose included fruit colour, overall fruit
shape and fruit shape at the basal end and apex.
3.2.1.5 Seed morphology
Seed colour and seed shape for different genotypes were
recorded at the mature stage of fruit.
3.2.2 Physical traits
3.2.2.1 Leaf
Ten fully developed leaves from current growth of each plant
were selected at random at the time of fruit maturity from the
outside branches at the middle of canopy for the purpose of data
recording. Leaf length, leaf width and leaf area for different
genotypes were measured with the help of leaf area meter (AM
100, Analytical Development Company Ltd. England) and the
averages were taken for the purpose of analysis.
62
3.2.2.2 Inflorescence
Five clusters of each plant were selected at random from the
outside branches at the middle of the canopy to study the floral
characteristics. Length of panicle and number of flowers per
panicle was noted at the time of full bloom. Average values were
used for analysis. Number of days from flowering (when 5
percent flowers blossomed) to full bloom (when 70 percent of
the flowers on the trees fully opened) was also recorded as
described by Durgac et al., 2006.
3.2.2.3 Fruit
Fruit characteristics at mature stage were recorded from 20 fruits
randomly selected from different sides from middle of canopy.
Parameters included length of fruit, width of fruit, width / length
index, fruit weight, fruit volume, flesh to seed ratio by weight
and flesh / seed ratio by volume. Number of fruits per bunch was
calculated by taking the average of 10 bunches from each tree
randomly selected on the four sides at the middle of canopy.
Fruit yield per plant was recorded in kg. The period from full
bloom to maturity was also noted. The time when the greenness
of the fruits completely disappeared was considered as the
mature stage according to Badenes et al., 2000.
63
Fruit length and fruit width were recorded with the help of
‘vernear calipers’. Fruit width / length index was measured by
dividing fruit width by fruit length. Fruit weight was measured
with the help of electric balance.
Flesh to seed ratio by weight was calculated with the help of
following formula:
(Fruit weight – Seeds weight) Flesh to seed ratio by weight = --------------------------------------- Seeds weight
Flesh to seed ratio by volume was calculated by measuring the
fruit volume and seeds volume by water displacement method
using the following formula:
: (Fruit volume – Seeds volume) Flesh to seed ratio by volume = ------------------------------------- Seeds volume
3.2.2.4 Seed
Seeds from 20 berries randomly selected from middle of plant
canopy were used to observe the seed characteristics including
number of seeds per fruit, weight of individual seed and seed
content per fruit.
Number of seeds per fruit was calculated by counting the total
seeds contained by 20 berries and then taking the average. Seed
weight was noted by dividing the total seeds’ weight by the total
64
number of seeds. Seed content per fruit was worked out by the
following formula:
Seed content per fruit = Total seeds’ weight of 20 berries ÷ 20
3.2.3 Data Analyses
Statistical analysis of the physical traits was carried out in
randomized complete block design (RCBD) combined over years in
MSTAT-C package (MSTAT, 1991) and the means were compared by
Duncan’s Multiple Range test at 5% level of significance (Gomez and
Gomez, 1984).
3.3 DNA Polymorphism Analyses
For molecular marker studies, DNA was also isolated from leaf samples
and analyzed for Random Amplification Polymorphic DNA (RAPD) to assess the
genetic similarity / diversity among different cultivars of loquat. The laboratory
studies were carried out in the Department of Horticulture and the Department of
Biochemistry, Pir Mehr Ali Shah Arid Agriculture University Rawalpindi. Loquat
genotypes were characterized on the basis of morpho-physical traits and molecular
markers.
Loquat genotypes at all the selected locations representing genetic
diversity on the basis of morphophysical characteristics were subjected to
RAPD analysis. For this purpose tender leaves from the subject plants were
collected and DNA extraction was done by the method described by Zidani
65
et al. (2005). The detailed procedure followed for RAPD analysis is given
below:
3.3.1 DNA Extraction
Leaves were harvested and frozen immediately in liquid nitrogen. A 0.3 g
of leaf sample was ground in liquid nitrogen using a mortar and pestle. The
pulverized leaves were transferred to centrifuge tube. CTAB buffer (2%)
containing 1% (v/v) -mercaptoethanol and 1% PVP was then added to the tube
and thoroughly mixed. The tube was incubated at 60°C for 30 min with frequent
swirling. An equal volume of chloroform: isoamylalcohol (24:1) was added and
centrifuged at 10,000 rpm at 4°C for 15 min to separate the phases. The
supernatant was decanted and transferred to a new tube. The above mentioned
steps, starting with the addition of chloroform: isoamylalcohol (24:1) and ending
with decanting of supernatant was repeated twice.
The supernatant was precipitated with 2/3 volume of ethanol. The
precipitated nucleic acids were collected and washed twice with the buffer (75%
ethanol, 3 M sodium acetate, TE). Care was taken not to shake the tube vigorously
because DNA is very vulnerable to fragmentation at this stage. The pellet was air
dried and suspended in TE. The dissolved nucleic acids were brought to 1.4 M
NaCl and re-precipitated using 2 volumes of 75% ethanol. The pellet was washed
using 100 % ethanol. It was then dried and re-suspended in 100 µl of TE buffer.
The tube was incubated at 37°C for 30 min to dissolve genomic DNA, and RNase
was then added. Thereafter, the quantification of the DNA was done by
66
measuring optical density (OD) at 260 and 280 nm using
spectrophotometer.
3.3.2 Agarose Gel Electrophoresis
Quality of the extracted DNA was checked by running on 1.5 %
agarose gel. Ethidium bromide solution was used to stain the gel. Stained
gel was visualized by UV transilluminator and quality of the DNA was
assessed. The appropriate dilutions of DNA were made for the further
amplification and RAPD analysis.
3.3.3 Polymerase Chain Reaction
Polymerase chain reaction (PCR) was performed on T-Cy Thermal
cycler (Crea Con, The Netherlands) using 25 l reaction mixture
containing 20 mM Tris-HCl (pH 7.8), 100 mM KCl, 3 mM MgCl2, 200 µM
of each dNTP, 1µM Primer, 50 ng of DNA and one unit of Taq DNA
polymerase. Fourteen 10 base pair primers were used for amplification
reactions (Table 2), however, only the primers showing polymorphic
results were selected for the purpose of diversity and similarity analysis.
After PCR, the amplified fragments were separated on 1.5 % agarose
gel in 0.5 X Tris Boric EDTA (TBE) buffer, stained by ethidium bromide,
visualized and photographed with the help of gel documentation system
(Kodak EDAS 290).
67
Table 2 Ten base pair primers used for the DNA amplification of loquat
S. No. Name of the Primer Sequence (5- 3)
1 GL DecamerA-01 GAGGCCCTTC
2 GL DecamerA-02 TGCCGAGCTG
3 GL DecamerA-05 AGGGGTCTTG
4 GL DecamerA-06 GGTCCCTGAC
5 GL DecamerA-09 GGGTAACGCC
6 GL DecamerC-02 GTGAGGCGTC
7 GL DecamerC-05 GATGACCGCC
8 GL DecamerC-07 GTCCCGACGA
9 GL DecamerC-08 TGGACCGGTG
10 GL DecamerC-09 CTCACCGTCC
11 GL DecamerC-17 TTCCCCCCAG
12 GL DecamerC-18 TGAGTGGGTG
13 GL DecamerC-19 GTTGCCAGCC
14 GL DecamerC-20 ACTTCGCCAC
68
3.3.4 RAPD Data Analyses
Data recorded was statistically analyzed. The photographs of gels
were used to score data for RAPD markers. RAPD behave as dominant
markers (Clark and Lanigan, 1993), thus they tend to be bi-state (present-
absent) type of scoring. Each DNA fragment amplified by a given primer
was considered as a unit character and the RAPD fragments were scored as
present (1) or absent (0) for each of the primer-accession combinations.
The molecular size of the amplification products was measured with
marker DNA ladder. The presence and absence of the bands was scored in
a binary data matrix. Polymorphic bands were scored and used for further
analysis. RAPD analysis was carried out using the Statistica 5.5 software.
Dandrogram was constructed showing the degree of related / differences among all
the genotypes.
69
Chapter 4
RESULTS AND DISCUSSION
Loquat is an important fruit crop of some areas of Pakistan. It gives good
returns to the growers, as there is no other fresh fruit available in the market during
March / April. Unfortunately this fruit did not attract the attention of the
researchers in the past. Although there are a number of genotypes scattered in
different areas of the loquat-growing region, but no work describing characteristics
of these genotypes has been reported so far in the country. Loquats are mainly
propagated through seed, so it has led to development of many new accessions,
which are result of different crosses occurring in the orchards naturally. Concept of
only two cultivars exists in the farming community; the red flesh cultivar locally
known as ‘Ratta’ and the white flesh cultivar known as ‘Saita’. The same concept
has been presented by Genai (1968). Actually these are not two cultivars but two
loquat groups based on their flesh colour, each group comprising of a number of
genotypes with a variety of characteristics.
Better genotypes exist rarely in some orchards of the remote areas but most
of the orchards are stuffed with the inferior genotypes having poor quality fruit.
Since no screening of better genotypes has been done in the past, hence no special
attention has been given towards their conservation or multiplication and they are
at the verge of extinction. Study regarding the characterization of loquat genotypes
has been performed for the first time in Pakistan. One aspect of this work includes
the study of different loquat genotypes available at all the selected locations in the
loquat growing region on the basis of morpho-physical traits. The other aspect
70
consists of the characterization on the basis of some RAPD markers and gives a
view of the genetic diversity and similarity of the loquat genotypes at DNA level.
4.1 CHARACTERIZATION OF LOQUAT GENOTYPES ON THE
BASIS OF MORPHO-PHYSICAL TRAITS
4.1.1 Kalar Kahar
4.1.1.1 General morphology
All the genotypes had the semi upright tree habit except KK1, which had
the upright habit. KK2, KK3 and KK4 had the leaves with blunt acute tips while
the other two genotypes’ leaves had sharp acute tip. Shape of the panicle in all the
genotypes was conical (Table 3). In Spain, loquat varieties, ‘Cardona’ and
‘Italiano-1’ have been reported to have an upright and semi upright tree habit
respectively, while shape of panicle in both the varieties was conical (Llacer et al.,
2003).
4.1.1.2 Fruit and seed morphology
Skin colour ranged from yellowish white (KK5) to orange yellow (KK1,
KK2 and KK3) while pulp colour in all the genotypes was orange except in KK4,
which had yellowish white pulp colour. Fruit shape was round in KK3 and KK5,
obovoid in KK1 and KK4 while oblong in KK2. Fruit shape of all the genotypes at
the basal end was obtuse except that in KK4, which was round. Fruit shape at the
apex in case of KK2, KK3 and KK5 was raised. It was flat in KK1 while depressed
in KK4. Colour of seeds in KK1 was brown, whereas it was light brown in all
71
other genotypes. Seed shape in all the genotypes was the same i.e., elliptic
(Table 4).
In China, orange yellow skin as well as flesh colour has also been reported
in ‘Yangmeizhou 4’ (Wu, 2001), while orange skin as well as pulp colour has been
observed in ‘Kumquat loquat’ and ‘Sour loquat’ (Huang et al., 2007). In
‘Qingbian’, skin colour is yellowish white, whereas pulp colour is white (He et al.,
2007). ‘Ninghaibai’ has a yellowish white skin while the flesh colour is white with
a long round or round fruit shape (Feng et al., 2004). Round fruit shape has been
observed in ‘Donghuzao’ (Zhao et al., 2001), ‘Baili’ and ‘Qingzhong’ (Feng et al.,
2007). In Spain, Magdal and ‘Cardona’ cultivars have fruits with obovoid and
oblong shape respectively, while shape of seeds in both the cultivars has been
elliptic (Llacer et al., 2003).
4.1.1.3 Fruit characteristics
Fruit characteristics of the genotypes are given in Table 5. KK2 was at the
top with reference to fruit length (3.45 cm), fruit width (3.15 cm), fruit weight
(17.29 g), fruit volume (16.52 mm3), flesh seed ratio by weight (2.59), flesh seed
ratio by volume (2.91) and fruit yield per plant (49.03 kg). It was significantly
different from the genotypes which followed it. In all these parameters, KK3 has
been found at the bottom except for fruit yield per plant and fruit width. Maximum
number of fruits per bunch was observed in KK3 (16.27) followed by KK4
(15.05). Least fruit width (2.30 cm) as well as the width length index (0.77) was
observed in KK4. KK5 had the highest width length index (1.06) followed by
72
Table 3 General appearance of the loquat plants of 5 genotypes at Kalar Kahar
Genotypes Tree habit Shape of leaf tip Shape of panicle KK1 Upright Sharp acute Conical KK2 Semi upright Blunt acute Conical KK3 Semi upright Blunt acute Conical KK4 Semi upright Blunt acute Conical KK5 Semi upright Sharp acute Conical
Table 4 Fruit and seed morphology of 5 loquat genotypes at Kalar Kahar Genotypes Skin
colour Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
KK1 Orange yellow
Orange Obovoid Obtuse Flat Brown Elliptic
KK2 Orange yellow
Orange Oblong Obtuse Raised Light brown
Elliptic
KK3 Orange yellow
Orange Round Obtuse Raised Light brown
Elliptic
KK4 Yellow Yellowish white
Obovoid Round Depressed Light brown
Elliptic
KK5 Yellowish white
Orange Round Obtuse Raised Light brown
Elliptic
73
KK3 (0.99), while KK5 was at the bottom with reference to number of fruits per
bunch (8.83) and yield per plant (30.75 kg). KK3 took maximum days (131.00)
from full bloom to maturity. KK4 required the least time (115.5 days) from full
bloom to maturity.
KK2 was found to be the best among the 5 genotypes with reference to
most of the characteristics (fruit weight, fruit volume, flesh seed ratio by weight,
flesh seed ratio by volume and yield per tree). Its fruit weight (17.29 g) was
comparable with ‘Kumquat loquat’ which had a fruit weight of 18.0 g (Huang et
al., 2007) and even greater than ‘Taishan Zhong’ having 12.8 g fruit weight (He et
al., 2007). But it is very small as compared with the fruit weight observed in
‘Zhaozhong’ (30 g), ‘Jidanbai’ (35.6 g) and ‘Guangyu’ (43.61 g), which are some
of the main cultivars of China (Feng et al., 2007). In Spain, Buenet’, ‘Cardona’,
‘Peluches’ and ‘Tanaka’ cultivars have a fruit weight of 43.10 g, 45.40 g, 95.00 g
and 60.60 g respectively (Llacer et al., 2003). ‘Selezione 2 PA’, ‘Nespolone di
Trabia’ and ‘Ferdinando’ in Italy produce fruits weighing 43.30 g, 50.40 g and
44.20 g respectively (Insero et al., 2003).
Flesh seed ratio in KK2 (2.59) is comparatively low as compared with
many cultivars growing in China, Spain and Turkey. ‘Gold Nugget’, ‘Baffico’ and
‘Kanro’ in Turkey have been found to have a flesh seed ratio of 3.83, 4.16 and 5.42
respectively (Durgac et al., 2006). In Italy, flesh seed ratio in ‘Vainiglia’,
‘Ferdinando’ and ‘Magdal’ have been found to be 5.4, 5.3 and 6.5 respectively
74
Table 5 Fruit characteristics of 5 loquat genotypes at Kalar Kahar
Fruit
characteristics Year KK1 KK2 KK3 KK4 KK5
CV
%
Fruit length
(cm)
Year I 2.98 b 3.45 a 2.71 c 2.96 b 2.77 c 2.83
Year II 2.94 b 3.45 a 2.74 c 3.04 b 2.81 c 1.76
Mean 2.96 B 3.45 A 2.73 C 3.00 B 2.79 C 2.35
Fruit width
(cm)
Year I 2.43 d 3.15 a 2.69 c 2.28 e 2.94 b 2.37
Year II 2.34 d 3.14 a 2.71 c 2.33 d 3.00 b 2.71
Mean 2.39 D 3.15 A 2.70 C 2.30 D 2.97 B 2.55
WLI
Year I 0.82 d 0.91 c 0.99 b 0.77 e 1.06 a 1.78
Year II 0.79 d 0.91 c 0.99 b 0.76 e 1.07 a 1.69
Mean 0.81 D 0.91 C 0.99 B 0.77 E 1.06 A 1.74
Fruit weight
(g)
Year I 15.78 a 17.34 a 9.81 b 10.52 b 15.23 a 8.51
Year II 15.30 b 17.24 a 10.25 c 10.85 c 15.53 b 5.87
Mean 15.54 B 17.29 A 10.03 C 10.68 C 15.38 B 7.31
Fruit volume
(mm3)
Year I 15.06 ab 16.56 a 9.44 c 10.14 c 14.44 b 7.29
Year II 14.63 b 16.47 a 9.78 c 10.44 c 14.83 b 5.50
Mean 14.85 B 16.52 A 9.61 C 10.29 C 14.64 B 6.45
F:S wt.
Year I 2.41 b 2.59 a 1.93 c 2.02 c 2.38 b 4.26
Year II 2.40 b 2.60 a 1.99 c 2.07 c 2.40 b 3.31
Mean 2.40 B 2.59 A 1.96 C 2.04 C 2.39 B 3.81
F:S vol.
Year I 2.73 b 2.92 a 2.11 c 2.24 c 2.57 b 3.62
Year II 2.68 b 2.91 a 2.23 c 2.32 c 2.67 b 2.34
Mean 2.70 B 2.91 A 2.17 D 2.28 C 2.62 B 3.03
Fruits per
bunch
Year I 12.40 c 14.33 b 16.20 a 14.80 b 8.60 d 4.49
Year II 12.63 c 14.67 b 16.33 a 15.30 ab 9.07 d 4.15
Mean 12.52 C 14.50 B 16.27 A 15.05 B 8.83 D 4.32
Days FB to M
Year I 121.0 c 125.7 b 130.0 a 115,0 d 116.3 d 1.22
Year II 119.0 c 127.0 b 132.0 a 116.0 c 118.0 c 1.48
Mean 120.0 C 126.3 B 131.0 A 115.5 D 117.2 D 1.36
Yield per tree
(kg)
Year I 44.23 ab 48.50 a 37.95 c 40.26 bc 32.22 d 6.57
Year II 45.47 ab 49.57 a 36.49 c 41.09 bc 29.27 d 7.44
Mean 44.85 B 49.03 A 37.22 C 40.67 C 30.75 D 7.02
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by volume ; Days FB to M = Days from full bloom to maturity
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
75
(Insero et al., 2003). High flesh seed ratios were observed in ‘Cardona’, Buenet’,
‘Peluches’ and ‘Tanaka’ (6.20, 7.08, 7.48 and 5.38 respectively) at Spain (Llacer et
al., 2003).
Fruit yield of KK2 (49.03 kg / tree) is much better than that of ‘Kanro’
which yielded 24.5 kg / tree, while it was very low as compared with ‘Champagne
de Grasse’ (Karadeniz, 2003) and ‘Algerie’ (Hermoso and Farre, 2003), which
gave 70 kg and 74 kg fruit per tree respectively.
4.1.1.4 Seed characteristics
Maximum number of seeds per fruit (3.63) was observed in KK2 followed
by KK3 (3.45) both genotypes being statistically at par with each other. Lowest
number of seeds per fruit (2.14) was observed in KK5. On the other hand, KK5
had the maximum seed weight (2.12 g) followed by KK1 (1.49 g) while KK3 had
the lowest seed weight (0.99 g). Seed content per fruit was highest in KK2 (4.81 g)
followed by KK1 (4.56 g) and KK5 (4.54 g), all the three genotypes having non
significant difference. Lowest seed content (3.39 g) was observed in KK3. Seed
characteristics are given in Table 6.
‘Algerie’ in Spain had 2.30 seeds per fruit (Llacer et al., 2003), which is
slightly higher than that observed in KK5 (2.14), while in China, ‘Taicheng 4’ (Xie
et al., 2007) and ‘White loquat’ (Huang et al., 2007) were reported to have only
1.32 and 2 seeds per fruit respectively. ‘Peluches’ in Spain had 3.7 seeds per fruit
with 11.20 g seed content per fruit (Llacer et al., 2003), but also had a high flesh
seed ratio of 7.48 as its fruit is very large. In Italy, ‘Selezione 2 PA’, ‘Nespolone di
76
Trabia’ ‘Ferdinando’ and ‘Vainiglia’ were observed to have 3.2, 3.8, 3.4 and 3.7
seeds per fruit respectively (Insero et al., 2003).
4.1.1.5 Leaf characteristic
KK2 had the maximum leaf length (28.14 cm) followed by KK1 (27.91
cm) both being statistically non significant with each other. KK1 had the maximum
leaf width (9.67 cm) followed by KK2 (9.36 cm) while KK2 was at par with KK1
as well as with KK4. Leaf area was highest (167.7 cm2) in KK1 and was followed
by KK2 (166 cm2) with non significant difference. KK5 was at the bottom with
reference to leaf length (13.43 cm), leaf width (4.18 cm) as well as leaf area (39.47
cm2). Leaf characteristics are shown in Table 7. In China, ‘Hanwuzhong’ is a
loquat variety which has average leaf length of 26.50 cm and leaf width of 9.00 cm
(He et al., 2007) This leaf size is somewhat less than that found in KK1, KK2 and
KK4 while greater than that of the other two genotypes.
4.1.1.6 Inflorescence
Number of flowers per panicle was highest in KK4 (177.89) followed by
KK3 (173.93) with non significant difference. This number is comparable with
‘Madgal’ variety, which had 178 flowers per panicle in Spain (Llacer et al., 2003).
Number of flowers per panicle was significantly low in other genotypes. Least
number of flowers per panicle was observed in KK5 (113.41) which is comparable
with 108.10 flowers per panicle observed in ‘Nespolone di Trabia’ (Insero et al.,
2003). Size of panicle was largest in KK4 (20.72 cm). It was followed by KK2
(19.58 cm) with significant difference. The smallest panicle size (15.44 cm) was
77
Table 6 Seed characteristics of 5 loquat genotypes at Kalar Kahar
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean KK1 3.10 b 3.03 b 3.07 C 1.50 b 1.49 b 1.49 B 4.63 a 4.50 a 4.56 A
KK2 3.67 a 3.58 a 3.63 A 1.32 c 1.34 c 1.33 C 4.83 a 4.79 a 4.81 A
KK3 3.28 ab 3.62 a 3.45 AB 1.02 d 0.95 e 0.99 E 3.35 b 3.43 b 3.39 B
KK4 3.20 ab 3.23 b 3.22 BC 1.09 d 1.09 d 1.09 D 3.48 b 3.54 b 3.51 B
KK5 2.10 c 2.18 c 2.14 D 2.15 a 2.09 a 2.12 A 4.51 a 4.57 a 4.54 A
CV % 7.82 4.61 6.39 4.20 4.02 4.11 6.81 4.88 5.92
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
78
Table 7 Leaf characteristics of 5 loquat genotypes at Kalar Kahar
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2) Year I Year II Mean Year I Year II Mean Year I Year II Mean
KK1 27.74 ab 28.08 a 27.91 A 9.70 a 9.65 a 9.67 A 170.0 a 165.4 a 167.7 A KK2 28.05 a 28.24 a 28.14 A 9.48 a 9.23 a 9.36 AB 168.2 a 163.9 a 166.0 A KK3 21.38 c 20.89 c 21.14 C 7.31 b 7.33 b 7.32 C 108.6 c 106.8 b 107.7 C KK4 26.89 b 26.24 b 26.57 B 9.38 a 8.99 a 9.19 B 158.3 b 154.3 a 156.3 B KK5 13.77 d 13.08 d 13.43 D 4.21 c 4.16 c 4.18 D 40.16 d 38.78 c 39.47 D CV % 2.14 3.12 2.67 3.66 5.14 4.45 3.94 5.66 4.85
79
Table 8 Floral characteristics of 5 loquat genotypes at Kalar Kahar
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full bloom
Year I Year II Mean Year I Year II Mean Year I Year II Mean
KK1 147.08 b 153.44 b 150.26 B 16.49 c 16.86 b 16.67 C 41.33 ab 42.67 ab 42.00 B
KK2 149.19 b 154.85 b 152.02 B 19.40 b 19.76 a 19.58 B 40.00 b 40.33 b 40.17 C
KK3 171.21 a 176.65 a 173.93 A 16.54 c 16.95 b 16.74 C 43.33 a 44.67 a 44.00 A
KK4 175.78 a 179.99 a 177.89 A 21.17 a 20.26 a 20.72 A 39.33 b 37.00 c 38.17 D
KK5 111.35 c 115.47 c 113.41 C 15.19 d 15.69 b 15.44 D 39.67 b 41.00 b 40.33 BC
CV % 4.41 5.64 5.08 3.69 3.76 3.73 3.07 3.63 3.37
80
found in KK5. The genotype KK3 took maximum time (44 days) from flowering
to full bloom while KK1 took 42 days. Number of days from flowering to full
bloom was lowest (38.17 days) in KK4 (Table 8).
4.1.2 Choa Saiden Shah
4.1.2.1 General morphology
All the genotypes had the spreading tree habit with sharp acute shape of
leaf tips. Shape of panicle in all the three genotypes was also the same that is
conical (Table 9). Loquat cultivar, ‘Peluches’ in Spain has spreading tree habit
with a conical shape of panicles. ‘Buenet’, ‘Saval-2’ and ‘Crisanto Amadeo’ also
have conical shape of panicle, while tree habit in these cultivars is upright (Llacer
et al., 2003).
4.1.2.2 Fruit and seed morphology
Skin colour in three genotypes was orange yellow. Pulp colour in CS1 and
CS3 was yellowish white while it was orange in CS2. Fruit shape was obovoid in
CS1 and CS2 while oblong in CS3. Fruit shape of all the three genotypes at the
basal end was obtuse. Fruit shape at the apex in case of CS1 and CS2 was raised
while it was flat in CS3. Seed colour in CS1 and CS2 was light brown while it was
brown in CS3. Seed shape in all the genotypes was the same i.e., elliptic (Table
10).
In Spain, orange yellow skin as well as pulp colour has been observed in a
number of loquat varieties including ‘Cardona’, ‘Algerie’ and ‘Golden Nugget’.
81
Table 9 General appearance of loquat plants of 3 genotypes at Choa
Saiden Shah
Genotypes Tree habit Shape of leaf tip Shape of panicle
CS1 Spreading Sharp acute Conical
CS2 Spreading Sharp acute Conical
CS3 Spreading Sharp acute Conical
Table 10 Fruit and seed morphology of 3 loquat genotypes at Choa
Saiden Shah
Genotypes
Skin colour
Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
CS1 Orange yellow
Yellowish white
Obovoid Obtuse Raised Light brown
Elliptic
CS2 Orange yellow
Orange Obovoid Obtuse Raised Light brown
Elliptic
CS3 Orange yellow
Yellowish white
Oblong Obtuse Flat Brown Elliptic
82
‘Buenet’ has orange skin as well as pulp colour (Llacer et al., 2003). ‘Qingbian’ in
China has a yellowish white skin colour and white pulp colour (He et al., 2007).
‘Madgal’, ‘Golden Nugget’ and ‘Tanaka’ have fruits with obovoid shape, while
‘Algerie’ and ‘Buenet’ have fruits with oblong shape. Elliptical seed shape has
been reported in ‘Magdal’, ‘Algerie’ and ‘Cardona’ (Llacer et al., 2003).
4.1.2.3 Fruit characteristics
Fruit characteristics of the genotypes are given in Table 11. Fruit length
was highest in CS2 (3.62 cm) which was followed by CS1 (3.52 cm) both being at
par. CS3 had significantly low fruit length (3.03 cm). Fruit width was significantly
highest in CS2 (3.21 cm) followed by CS1 (2.70 cm). It was least in CS3 (2.64 cm)
which was at par with CS1. Width length index was maximum in CS2 (0.89)
followed by CS3 (0.87) and was significantly low in CS1 (0.77). Fruit weight was
highest in CS2 (21.37 g) followed by CS1 (15.42 g) and minimum in CS3 (11.47
g) showing significant difference in all the three genotypes. Fruit volume was also
highest in CS2 (20.61 mm3) and lowest in CS3 (10.81 mm3). Flesh seed ratio by
weight was highest in CS1 (2.55) followed by CS2 (2.46) both genotypes being at
par. Flesh seed ratio by volume was also highest in CS1 (2.94) followed by CS2
(2.86) both being at par. CS3 had significantly low flesh seed ratio by weight
(2.04) and flesh seed ratio by volume (2.31). Significant differences were observed
in terms of number of fruits per bunch. It was maximum in CS1 (16.07) and
minimum in CS3 (11.92). CS3 took maximum time from full bloom to maturity
(127.83 days) while this time was significantly low in CS1 (120.33 days).
83
Table 11 Fruit characteristics of 3 loquat genotypes at Choa
Saiden Shah
Fruit
characteristics
Year CS1 CS2 CS3 CV %
Fruit length
(cm)
Year I 3.48 a 3.61 a 2.99 b 4.70
Year II 3.55 a 3.63 a 3.06 b 3.93
Mean 3.52 A 3.62 A 3.03 B 4.33
Fruit width
(cm)
Year I 2.67 b 3.20 a 2.62 b 4.63
Year II 2.73 b 3.21 a 2.65 b 4.05
Mean 2.70 B 3.21 A 2.64 B 4.35
WLI Year I 0.77 b 0.89 a 0.87 a 2.73
Year II 0.77 b 0.88 a 0.87 a 1.43
Mean 0.77 B 0.89 A 0.87 A 2.12
Fruit weight
(g)
Year I 15.21 b 21.13 a 11.31 c 9.04
Year II 15.63 b 21.60 a 11.62 c 3.75
Mean 15.42 B 21.37 A 11.47 C 6.86
Fruit volume
(mm3)
Year I 14.61 b 20.38 a 10.68 c 9.24
Year II 14.96 b 20.84 a 10.95 c 4.17
Mean 14.79 B 20.61 A 10.81 C 7.11
F:S wt. Year I 2.53 a 2.44 a 2.00 b 7.49
Year II 2.56 a 2.48 a 2.07 b 4.52
Mean 2.55 A 2.46 A 2.04 B 6.15
F:S vol. Year I 2.95 a 2.82 a 2.29 b 8.50
Year II 2.92 a 2.89 a 2.34 b 3.26
Mean 2.94 A 2.86 A 2.31 B 6.41
Fruits per
bunch
Year I 16.30 a 12.33 b 11.70 b 8.86
Year II 15.83 a 12.83 b 12.13 b 8.17
Mean 16.07 A 12.58 B 11.92 B 8.52
Days FB to M Year I 121.33 b 126.33 ab 129.00 a 0.77
Year II 119.33 b 124.00 b 126.67 a 1.66
Mean 120.33 B 125.17 A 127.83 A 1.28
Yield per tree
(kg)
Year I 46.48 a 37.52 b 40.62 b 4.76
Year II 48.53 a 39.05 b 43.65 ab 5.98
Mean 47.51 A 38.28 C 42.13 B 5.44
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by
volume; Days FB to M = Days from full bloom to maturity
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
84
Significant differences were noted in terms of yield per tree. It was maximum in
CS1 (47.51 kg) and minimum in CS2 (38.28 kg). CS2 was the best one with
reference to fruit weight (21.37 g), but had the lowest yield per plant (38.28 kg).
Fruit weight of CS2 (21.37 g) is almost the same as that of ‘Maomu’
variety (21.70 g) in China (Luo, 2005) and ‘Baffico’ (22.55 g) in Turkey (Durgac
et al., 2006), but it is smaller than that of most of the other varieties growing in
Chian, Spain and Turkey. ‘Qingbian’, ‘Hanwuzhong’ and ‘Mojia No. 1’ in China
have fruit weight of 28.70 g, 30.97 g and 53.20 g respectively (He et al., 2007).
Fruit weight of ‘Magdal’ and ‘Crisanto Amadeo’ in Spain has been observed as
45.50 g and 68.70 g respectively (Llacer et al., 2003). In Turkey, ‘Ottawiani’ and
‘Dr. Trabut’ have been found to have fruit weight of 49.78 g and 43.23 g
respectively (Yalcin and Paydas, 1995). ‘Selezione 2 PA’. ‘Nespolone di Trabia’
and ‘Ferdinando’ in Italy produce large fruits weighing 43.30 g, 50.40 g and 44.20
g respectively (Insero et al., 2003).
CS1 proved to have the highest flesh seed ratio by weight (2.49) and flesh
seed ratio by volume (2.92). But its flesh seed ratio was very low as compared with
that of ‘Selezione 2 PA’ (4.8), ‘Ferdinando’ (5.3) and ‘Algerie’ (6.2) as observed
in Italy (Insero et al., 2003), ‘Kanro’ (5.42) and ‘Bafico’ (4.16) as noted in Turkey
(Durgac et al., 2006). High flesh seed ratios were observed in ‘Cardona’, Buenet’,
‘Peluches’ and ‘Tanaka’ (6.20, 7.08, 7.48 and 5.38 respectively) in Spain (Llacer et
al., 2003). In another study conducted in Turkey, 13 types of loquat, b1 to b13,
were reported to have flesh seed ratios ranging from 3.88 to 5.10 (Polat, 2007).
85
Best yield per plant among 3 genotypes (47.51 kg) noted in CS1 is higher
than ‘Kanro’ (24.5 kg per plant), while lower than ‘Champagne de Grasse’, (70 kg
per plant) ‘M. Marie’ (69 kg per plant) in Turkey (Karadenez, 2003) and ‘Algerie’
(74 kg per plant) in Spain (Hermoso and Farre, 2003).
4.1.2.4 Seed characteristics
Significant differences were observed in terms of number of seeds per fruit.
Maximum number of seeds per fruit was observed in CS1 (3.64) followed by CS3
(3.39) with a significant difference. CS2 had the lowest number of seeds per fruit
(3.28) and was at par with CS3. Significant differences were noted in weight per
seed and seed content per fruit. Weight per seed was maximum in CS2 (1.89 g)
followed by CS1 (1.20 g) and minimum in CS3 (1.11 g). Similarly, seed content
per fruit was highest in CS2 (6.18 g) and lowest in CS3 (3.78 g). Table 12 shows
the seed characteristics of the three genotypes.
The genotype with the biggest fruit size (CS2) has been observed to have
the lowest number of seeds per fruit (3.28), while highest seed content per fruit
(6.18 g). Number of seeds per fruit and seed content per fruit observed in CS2 are
low as compared with those of ‘Magdal’ (3.70 seeds per fruit and 7.80 g seed
content), observed in Spain. But fruit weight of ‘Magdal’ is more than double the
weight of CS2 (Llacer et al., 2003). In Italy, ‘Selezione 2 PA’, ‘Nespolone di
Trabia’ ‘Ferdinando’ and ‘Vainiglia’ were observed to have 3.2, 3.8, 3.4 and 3.7
seeds per fruit respectively (Insero et al., 2003). In China, ‘Taicheng 4’ (Xie et al.,
2007) and ‘White loquat’ (Huang et al., 2007) were reported to have only 1.34 and
2 seeds per fruit respectively.
86
4.1.2.5 Leaf characteristic
Leaf characteristics of the three genotypes are given in Table 13.
Differences were non significant in terms of leaf length while significant in terms
of leaf width. Maximum leaf width was observed in CS2 (9.06 cm) followed by
CS1 (8.72 cm). CS3 had the minimum leaf width (8.30 cm) with significant
difference. Significant differences were also noted in case of leaf area. Leaf area
was maximum in CS2 (148.33 cm2) followed with a significant difference by CS3
(126.71 cm2) while it was minimum in CS1 (120.40 cm2). ‘Ningbai 1’ in China has
almost the same leaf size as found in the above genotypes. In this cultivar, leaf
length and width have been observed as 25.5 cm and 8.00 cm respectively (Feng et
al., 2007).
4.1.2.6 Inflorescence
Significant differences were observed in terms of floral characteristics
(Table 14). CS1 had the maximum number of flowers per panicle (157.68) and
maximum panicle size (20.47 cm). CS3 had the minimum number of flowers per
panicle (144.40) and minimum panicle size (18.31 cm). CS3 took maximum time
from flowering to full bloom (39.00 days) while this time was least in case of CS2
(34.50 days). In China, number of flowers per panicle has been noted as 61 in
‘Ningbai 2’ (Feng et al., 2007) and 134 in ‘Luoyangqing’ (Yang et al., 2007).
87
Table 12 Seed characteristics of 3 loquat genotypes at Choa Saiden Shah
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
CS1 3.72 a 3.57 a 3.64 A 1.16 b 1.23 b 1.20 B 4.30 b 4.39 b 4.35 B
CS2 3.23 b 3.32 b 3.28 B 1.90 a 1.87 a 1.89 A 6.14 a 6.21 a 6.18 A
CS3 3.37 b 3.42 b 3.39 B 1.12 b 1.11 c 1.11 C 3.77 c 3.78 c 3.78 C
CV % 4.27 2.22 3.40 4.44 2.07 3.46 4.07 1.94 3.18
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
Table 13 Leaf characteristics of 3 loquat genotypes at Choa Saiden Shah
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2)
Year I Year II Mean Year I Year II Mean Year I Year II Mean CS1 22.07 ns 21.61 ns 21.84 ns 8.76 a 8.68 ab 8.72 B 122.30 b 118.51 b 120.40 B
CS2 21.82 ns 22.52 ns 22.17 ns 9.02 a 9.10 a 9.06 A 146.66 a 150.01 a 148.33 A
CS3 22.56 ns 22.00 ns 22.28 ns 8.35 b 8.24 b 8.30 C 128.49 b 124.94 b 126.71 B
CV % 3.05 3.72 3.40 1.61 2.39 2.03 5.32 5.41 5.37
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
88
Table 14 Floral characteristics of 3 loquat genotypes at Choa Saiden Shah Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full bloom
Year I Year II Mean Year I Year II Mean Year I Year II Mean
CS1 155.67 ns 159.68 a 157.68 A 20.28 a 20.67 a 20.47 A 37.00 a 38.67 a 37.83 A
CS2 146.70 ns 151.13 ab 148.91 B 19.20 ab 19.25 b 19.23 B 34.33 b 34.67 b 34.50 B
CS3 143.27 ns 145.53 b 144.40 B 18.14 b 18.47 c 18.31 C 38.00 a 40.00 a 39.00 A
CV % 2.65 2.30 2.48 3.65 1.68 2.83 2.42 4.41 3.59
Means not sharing a letter differ significantly at p < 0.05 Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
89
4.1.3 Chhattar
4.1.3.1 General morphology
All the genotypes had the upright tree habit and conical shape of the
panicle. Shape of leaf tip was similar i.e., blunt acute in CH1 and CH2. In case of
CH3, shape of leaf tip was sharp acute (Table 15). ‘Algerie’, ‘Tanaka’, ‘Buenet’
and ‘Saval 2’ cultivars of loquat cultivars have also been reported to have upright
tree habit with conical shape of panicles in Spain (Llacer et al., 2003).
4.1.3.2 Fruit and seed morphology
Skin colour as well as the pulp colour in all the three genotypes was orange
yellow. Same skin and pulp colours have also been observed in ‘Msgdal’,
‘Cardona’, ‘Algerie’ and ‘Golden Nugget’ cultivars in Spain (Llacer et al., 2003),
but they have a larger fruit size as compared with that of the genotypes of Chhattar.
All these genotypes had different fruit shapes. Fruit shape was obovoid, round and
oblong in CH1, CH2 and CH3 respectively. Although, fruit shape at the basal end
in these genotypes was same i.e., obtuse, fruit shape at the apex was flat in CH1
while raised in case of other two genotypes. Seed colour was brown in CH1 and
light brown in the other two genotypes. All the genotypes were similar with respect
to seed shape which was elliptic (Table 16).
In Spain, elliptical seed shape has been observed in ‘Magdal’, ‘Cardona’
and ‘Saval-2’ while shape of fruits in these cultivars was obovoid, oblong and
round respectively (Llacer et al., 2003). In China round fruit shape has been
90
Table 15 General appearance of the loquat plants of 3 genotypes at
Chhattar
Genotypes Tree habit Shape of leaf tip Shape of panicle CH1 Upright Blunt acute Conical CH2 Upright Blunt acute Conical CH3 Upright Sharp acute Conical
Table 16 Fruit and seed morphology of 3 loquat genotypes at Chhattar
Genotypes Skin colour
Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
CH1 Orange yellow
Orange yellow
Obovoid Obtuse Flat Brown Elliptic
CH2 Orange yellow
Orange yellow
Round Obtuse Raised Light brown
Elliptic
CH3 Orange yellow
Orange yellow
Oblong Obtuse Raised Light brown
Elliptic
91
observed in ‘Donghuzao’ (Zhao et al., 2001), ‘Baili’ and ‘Qingzhong’ (Feng et al.,
2007). ‘Jidanbai’ and ‘Niuteibaisha’ had obovoid, while ‘Guangyu’ had oblong
fruit (Feng et al., 2007).
4.1.3.3 Fruit characteristics
Fruit characteristics of the genotypes are given in Table 17. Fruit length
was highest in CH3 (3.94 cm) followed by CH1 (3.54 cm) with significant
difference. It was lowest in CH2 (3.42 cm). Fruit width was highest in CH3 (3.40
cm) which was significantly higher than that of CH2 (3.23 cm). Lowest fruit width
was observed in CH1 (2.59 cm). Width length index was highest in CH2 (0.94)
followed by CH3 (0.87). CH1 had the lowest width length index. Significant
differences were also observed in terms of fruit weight as well as fruit volume.
Fruit weight was highest in CH3 (23.06 g) and lowest in CH1 (12.71 g). Similarly,
fruit volume was highest in CH3 (22.28 cm3) and lowest in CH1 (11.82 mm3).
CH3 and CH2 were at par with respect to flesh seed ratio by weight (2.68 and 2.66
respectively) and flesh seed ratio by volume (3.15 and 3.02 respectively). These
ratios were least in CH1 (1.68 and 1.92 respectively).
Maximum fruits per bunch were observed in CH2 (16.33) followed by CH1
(15.83) both being at par. CH3 had the lowest number of fruits per bunch (14.62).
CH1 took maximum time from full bloom to maturity (113.33 days). This period
was lowest in CH3 (103.83 days). Significant differences were observed in terms
of fruit yield per tree. Yield was highest in CH1 (40.66 kg) followed by CH3
(34.77 kg) and lowest in CH2 (28.13 kg) all having significant differences.
92
Although, among the three genotypes, CH1 had the highest yield, it was at the
bottom with reference to fruit weight. CH3 was found to be the best of the three
genotypes with respect to fruit weight, flesh seed ratio by weight and flesh seed
ratio by volume.
Fruit weight of CH3 is comparable with that of ‘Baffico’, which had a fruit
weight of 22.55 g, while smaller than ‘Dr. Tarbut’, which had a fruit weight of
29.54 g in Turkey (Durgac et al., 2006). Many loquat cultivars in China have the
fruits which are much larger than those of the genotypes of Chhattar. Fruit weight
was 30.97 g in ‘Hanwuzhong’, 53.2 g in ‘Mojia No. 1’ (He et al., 2007), 63.1 g in
‘Hongdenglong’ (Jiang et al., 2001) and 59.2 g in ‘Donghuzao’ (Zhao et al., 2001;
Zhao et al., 2003). In Spain, ‘Cardona’, ‘Algerie’, ‘Buenet’ and ‘Crisanto Amadeo’
have been observed to have fruit weight of 45.4 g, 65 g, 58.2 g and 68.7 g
respectively (Llacer et al., 2003). ‘Selezione 2 PA’. ‘Nespolone di Trabia’ and
‘Ferdinando’ in Italy have fruit weight of 43.30 g, 50.40 g and 44.20 g respectively
(Insero et al., 2003).
In most of the loquat cultivars grown in other loquat growing countries,
flesh seed ratios are very high than those found in the genotypes of Chhattar. In
Turkey, ‘Gold Nugget’, ‘Baffico’ and ‘Kanro’ in Turkey were found to have a
flesh seed ratio of 3.83, 4.16 and 5.42 respectively (Durgac et al., 2006). In a
recent study conducted in Turkey, 13 types of loquat, b1 to b13, were observed to
have flesh seed ratios ranging from 3.88 to 5.10 (Polat, 2007). In Italy, flesh seed
93
Table 17 Fruit characteristics of 3 loquat genotypes at Chhattar
Fruit
characteristics
Year CH 1 CH 2 CH 3 CV %
Fruit length
(cm)
Year I 3.52 b 3.42 c 3.93 a 1.32
Year II 3.55 b 3.43 c 3.94 a 0.95
Mean 3.54 B 3.42 C 3.94 A 1.15
Fruit width
(cm)
Year I 2.57 c 3.22 b 3.39 a 2.16
Year II 2.61 c 3.23 b 3.41 a 1.56
Mean 2.59 C 3.23 B 3.40 A 1.88
WLI Year I 0.73 c 0.94 a 0.87 b 1.52
Year II 0.73 c 0.94 a 0.86 b 1.04
Mean 0.73 C 0.94 A 0.87 B 1.31
Fruit weight
(g)
Year I 12.41 c 18.73 b 22.90 a 5.58
Year II 13.02 c 19.38 b 23.21 a 3.86
Mean 12.71 C 19.06 B 23.06 A 4.78
Fruit volume
(mm3)
Year I 11.57 c 17.66 b 22.15 a 5.88
Year II 12.08 c 18.34 b 22.42 a 3.92
Mean 11.82 C 18.00 B 22.28 A 4.97
F:S wt. Year I 1.66 b 2.65 a 2.70 a 3.34
Year II 1.70 b 2.66 a 2.66 a 3.64
Mean 1.68 B 2.66 A 2.68 A 3.49
F:S vol. Year I 1.92 b 3.02 a 3.17 a 3.12
Year II 1.92 b 3,02 a 3.13 a 4.02
Mean 1.92 B 3.02 A 3.15 A 3.59
Fruits per
bunch
Year I 15.70 a 16.13 a 14.50 b 3.24
Year II 15.97 ab 16.53 a 14.73 b 3.35
Mean 15.83 A 16.33 A 14.62 B 3.30
Days FB to M Year I 114.00 a 110.33 b 104.67 c 1.66
Year II 112.67 a 111.67 a 103.00 b 1.28
Mean 113.33 A 111.00 B 103.83 C 1.49
Yield per tree
(kg)
Year I 40.35 a 27.82 c 35.07 b 4.18
Year II 40.97 a 28.45 c 34.47 b 7.19
Mean 40.66 A 28.13 C 34.77 B 5.89
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by
volume; Days FB to M = Days from full bloom to maturity
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
94
ratio in ‘Vainiglia’, ‘Ferdinando’ ‘Peluche’ and ‘Magdal’ have been found to be
5.4, 5.3, 5.9 and 6.5 respectively (Insero et al., 2003). High flesh seed ratios were
observed in Buenet’, ‘Peluches’, ‘Cardona’ and ‘Tanaka’ (7.08, 7.48, 6.20 and 5.38
respectively) in Spain (Llacer et al., 2003).
Yield of these genotypes at Chhattar is much better than that of ‘Kanro’
which yielded 24.5 kg / tree, while it is very low as compared with ‘M. Marie’ and
‘Champagne de Grasse’ which gave 69 kg and 70 kg fruit / tree respectively in
Turkey (Karadeniz, 2003). This yield is also low as compared with that of
‘Algerie’ (74 kg per fruit) and ‘Gold Nugget’ (74 kg per fruit) observed in Spain
(Hermoso and Farre, 2003).
4.1.3.4 Seed characteristics
Table 18 shows that maximum number of seeds per fruit (3.16) was
observed in CH3 followed by CH2 (3.11) both genotypes being statistically at par
with each other. Lowest number of seeds per fruit (2.84) was observed in CH1,
which is slightly greater than ‘Niuteibaisha’ cultivar in China having 2.78 seeds
per fruit (Feng et al., 2007). In Italy, ‘Selezione 2 PA’, ‘Ferdinando’ and
‘Vainiglia’ were observed to have 3.2, 3.4 and 3.7 seeds per fruit respectively
(Insero et al., 2003). ‘Algerie’ in Spain had 2.30 seeds per fruit (Llacer et al.,
2003). In China, ‘White loquat’ (Huang et al., 2007) and ‘Taicheng 4’ (Xie et al.,
2007) were reported to have only 2 and 1.32 seeds per fruit respectively.
Highest seed weight (1.98 g) was found in CH3, while the other two
genotypes i.e. CH2 and CH1 were at par with each other having seed weight 1.68 g
95
and 1.67 g respectively. All the three genotypes were significantly different in
terms of seed content per fruit. It was highest in CH3 (6.26 g) and lowest in CH1
(4.74 g).
4.1.3.5 Leaf characteristic
It is evident from Table 19 that all the three genotypes were significantly
different in terms of leaf characteristics. CH2 was at the top with reference to leaf
length (28.66 cm), leaf width (8.67 cm) and leaf area (147.74 cm2). Its leaf size is
comparable with ‘Qingzhong’ having 29.20 cm and 8.40 cm leaf length and leaf
width respectively (Feng et al., 2007). CH1 had the lowest leaf length (23.44 cm),
leaf width (6.81 cm) and leaf area (103.90 cm2). This leaf size is quite comparable
with ‘Wugongbai’ cultivar having 23.00 cm and 6.80 cm length and width
respectively (Feng et al., 2007).
4.1.3.6 Inflorescence
Number of flowers per panicle was maximum in CH2 (171.21) which was
significantly higher than that in CH1 (157.21) and CH3 (151.69), the last two
being at par with each other. In Spain, loquat cultivars ‘Vainiglia’ and ‘Nespolone
di Ficarazzi’ have been reported to produce 176.20 and 158.40 flowers per panicle
respectively (Insero et al., 2003), which is comparable with the genotypes of
Chhattar. Maximum panicle size was noted in CH1 (16.32 cm) followed by CH3
(15.94 cm) with non significant difference. Panicle size was minimum in CH2
(14.12 cm) having a significant difference with the other two genotypes. CH1 took
96
Table 18 Seed characteristics of 3 loquat genotypes at Chhattar
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
CH 1 2.83 b 2.85 b 2.84 B 1.65 b 1.69 b 1.67 B 4.67 b 4.81 c 4.74 C
CH 2 3.07 a 3.15 a 3.11 A 1.67 b 1.68 b 1.68 B 5.13 b 5.30 b 5.21 B
CH 3 3.13 a 3.18 a 3.16 A 1.98 a 1.99 a 1.98 A 6.19 a 6.33 a 6.26 A
CV % 2.28 1.96 2.13 3.29 1.14 2.45 4.46 1.73 3.35
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
97
Table 19 Leaf characteristics of 3 loquat genotypes at Chhattar
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
CH 1 24.05 b 22.82 c 23.44 C 6.87 b 6.75 c 6.81 C 105.56 b 102.25 b 103.90 C
CH 2 28.31 a 29.01 a 28.66 A 8.64 a 8.70 a 8.67 A 146.25 a 149.22 a 147.74 A
CH 3 25.44 b 24.94 b 25.19 B 7.61 b 7.52 b 7.57 B 118.58 b 113.90 b 116.24 B
CV % 3.98 3.54 3.77 4.79 3.67 4.27 5.96 5.07 5.54
98
Table 20 Floral characteristics of 3 loquat genotypes at Chhattar
Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full bloom
Year I Year II Mean Year I Year II Mean Year I Year II Mean
CH 1 158.20 ab 156.22 b 157.21 B 16.47 a 16.17 a 16.32 A 45.33 a 46.67 a 46.00 A
CH 2 169.43 a 173.00 a 171.21 A 14.18 b 14.06 b 14.12 B 41.33 b 42.67 b 42.00 B
CH 3 149.60 b 153.78 b 151.69 B 16.03 a 15.85 a 15.94 A 42.00 ab 43.67 b 42.83 B
CV % 3.36 3.25 3.31 4.76 4.19 4.49 2.96 3.45 3.22
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
99
the maximum time from flowering to full bloom (46.00 days) which was
significantly higher than that in CH3 (42.83 days) and CH2 (42.00 days) while the
last two genotypes were at par with each other (Table 20).
4.1.4 Tret
4.1.4.1 General morphology
Genotypes TR1, TR2 and TR3 were found to have upright tree habit. Tree
habit was semi upright in TR4 and spreading in TR5. Shape of leaf tip was blunt
acute in TR1, TR3 and TR4 while it was sharp acute in TR2 and TR5. Shape of
panicle was truncate conical in TR4 while conical in all other genotypes (Table
21). In literature, upright, semi upright and spreading tree habit has been reported
in ‘Cardona’, ‘Italiano 1’, ‘Algerie’ and ‘Peluches’ cultivars of loquat respectively,
all having the same shape of panicles i.e. conical (Llacer et al., 2003).
4.1.4.2 Fruit and seed morphology
Fruit skin colour was yellowish white in TR3 while orange yellow in all
other genotypes. Pulp colour was yellowish white in TR3, orange in TR5 while it
was orange yellow in the other three genotypes. Fruit shape was obovoid in all the
genotypes except in TR5 which had oblong fruit shape. Fruit shape at the basal end
in all the genotypes was obtuse except in TR5 which had rounded fruit shape at the
basal end. Fruit shape at the apex was flat in TR1, TR4 and TR5 while raised in the
other two genotypes. Seed colour was brown in TR1 and TR5 while light brown in
the other genotypes. Seed shape was elliptic in genotypes TR1, TR3 & TR5 and
100
Table 21 General appearance of the loquat plants of 5 genotypes at Tret
Genotypes Tree habit Shape of leaf tip Shape of panicle
TR1 Upright Blunt acute Conical
TR2 Upright Sharp acute Conical
TR3 Upright Blunt acute Conical
TR4 Semi upright Blunt acute Truncate conical
TR5 Spreading Sharp acute Conical
Table 22 Fruit and seed morphology of 5 loquat genotypes at Tret
Genotypes Skin colour
Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
TR1 Orange yellow
Orange yellow
Obovoid Obtuse Flat Brown Elliptic
TR2 Orange yellow
Orange yellow
Obovoid Obtuse Raised Light brown
Round
TR3 Yellowish white
Yellowish white
Obovoid Obtuse Raised Light brown
Elliptic
TR4 Orange yellow
Orange yellow
Obovoid Obtuse Flat Light brown
Round
TR5 Orange yellow
Orange Oblong Rounded Flat Brown Elliptic
101
round in TR2 and TR4 (Table 22).
In Spain, elliptical seed shape has been reported in Magdal and ‘Cardona’
while round seed shape in ‘Golden Nugget’ and ‘Crisanto Amadeo’. The cultivas,
‘Madgal’, ‘Golden Nugget’ and ‘Tanaka’ were found to have obovoid fruits with
orange yellow skin as well as the pulp colour, whereas ‘Algerie’ and ‘Cardona’
had oblong fruit shape with orange yellow skin as well as the pulp colour (Llacer et
al., 2003). In China, ‘Baiyu’ and ‘Guangyu’ were reported to have oblong shaped
fruit (Feng et al., 2007).
4.1.4.3 Fruit characteristics
Fruit characteristics of the genotypes are given in Table 23. Significant
differences were observed among the different genotypes. Highest fruit length was
recorded in TR4 (5.09 cm) followed by TR2 (4.06 cm) with a significant
difference. It was lowest in TR5 (2.95 cm). Fruit width was also highest in TR4
(4.06 cm) followed by TR2 (3.10 cm) and lowest in TR5 (2.70 cm). Width length
index was highest in TR5 (0.91). It was followed with a significant difference by
TR1 (0.86) and TR3 (0.86), last two being at par. Width length index was least in
TR2 (0.76). Fruit weight was maximum in TR4 (38.77 g) followed by TR2 (20.04
g) with a significant difference. Minimum fruit weight was recorded in TR1 (12.73
g). Fruit volume was also maximum in TR5 (36.18 mm3) followed by TR2 (18.50
mm3) while minimum in TR5 (11.79 mm3).
TR4 was at the top with reference to flesh seed ratio by weight (2.80) and
flesh seed ratio by volume (3.03). It was followed by TR2 having these ratios as
102
2.67 and 2.89 respectively. TR3 was at the bottom with the lowest flesh seed ratio
by weight (2.11) as well as by volume (2.35). TR1 had the maximum number of
fruits per bunch (14.70) followed by TR4 (14.12) both being at par. Minimum
number of fruits per bunch was observed in TR5 (12.53) which was significantly
low among all the five genotypes. TR2 and TR3 took maximum time from full
bloom to maturity (121.00 and 121.50 days respectively) which was significantly
high as compared with the other genotypes. On the other hand, TR5 took the least
time from full bloom to maturity (112.83 days).
Yield per tree was highest in TR4 (54.93 kg) followed by TR3 (51.93 kg)
both being at par. TR5 had the lowest yield per tree (33.71 kg). All the genotypes
gave better yield as compared with that of ‘Kanro’ (24.50 kg per tree), while very
low as compared with ‘M. Marie’ (69 kg per tree) and ‘Champagne de Grasse’ (70
kg per tree) as observed in Turkey (Karadeniz, 2003). This yield is also low as
compared with that of ‘Algerie’ (74 kg per fruit) and ‘Gold Nugget’ (74 kg per
fruit) observed in Spain (Hermoso and Farre, 2003).
Among the 5 genotypes, TR4 was found to be the best one with reference
to fruit weight, flesh seed ratio by weight, flesh seed ratio by volume and yield per
tree. Its fruit weight (38.77 g) was much higher than that of ‘Dr. Trabut’ with an
average fruit weight of 29.54 g (Durgac et al., 2006) and ‘Hanwuzhong’ with a
fruit weight of 30.97 g (He et al., 2007). On the other hand, ‘Tanaka’ and
‘Peluches’ have even larger fruits, weighing 60.60 g and 95.00 g respectively
(Llacer et al., 2003). ‘Ottawiani’ and ‘Dr. Trabut’ have been found to have fruit
weight of 49.78 g and 43.23 g respectively (Yalcin and Paydas, 1995). ‘Selezione 2
103
PA’. ‘Nespolone di Trabia’ and ‘Ferdinando’ in Italy produce fruits weighing
43.30 g, 50.40 g and 44.20 g respectively (Insero et al., 2003). In Japan, leading
loquat cultivars, ‘Mogi’, ‘Nacasakiwase’ and ‘Tanaka’ have average fruit weights
of 50 g, 60 g and 70 g respectively (Durgac et al., 2006).
Studies conducted in leading loquat countries indicate that most of the
loquat cultivars grown over there have flesh seed ratios, which are very high than
those found in the genotypes of Tret. In Turkey, ‘Baffico’, ‘Gold Nugget’ and
‘Kanro’ in Turkey were found to have a flesh seed ratio of 4.16, 3.83, and 5.42
respectively (Durgac et al., 2006).
In another study, 13 types of loquat (b1 to b13), were observed to have
flesh seed ratios ranging from 3.88 to 5.10 (Polat, 2007). In Italy, flesh seed ratio
in ‘Ferdinando’, ‘Vainiglia’, ‘Peluche’ and ‘Magdal’ have been reported to be 5.3,
5.4, 5.9 and 6.5 respectively (Insero et al., 2003). In Spain, ‘Cardona’, Buenet’,
‘Peluches’ and ‘Tanaka’ had a high flesh seed ratio of 6.20, 7.08, 7.48 and 5.38
respectively (Llacer et al., 2003).
4.1.4.4 Seed characteristics
Significant differences were observed among the genotypes with respect to
the seed characteristics studied (Table 24). TR4 had the highest number of seeds
per fruit (4.83) followed by TR2 (4.23). Least number of seeds per fruit was
observed in TR1 (3.18). Maximum seed weight (2.11 g) as well as maximum seed
content per fruit (10.19 g) was also recorded in TR4. It was followed by TR2 in
these characteristics, having 1.29 g seed weight and 5.46 g seed content per fruit.
104
Table 23 Fruit characteristics of 5 loquat genotypes at Tret
Fruit
characteristics
Year TR1 TR2 TR3 TR4 TR5 CV
%
Fruit length
(cm)
Year I 3.18 d 4.08 b 3.40 c 5.07 a 2.93 e 2.30
Year II 3.15 d 4.04 b 3.37 c 5.10 a 2.97 e 2.10
Mean 3.17 D 4.06 B 3.39 C 5.09 A 2.95 E 2.20
Fruit width
(cm)
Year I 2.75 cd 3.13 b 2.91 c 4.03 a 2.68 d 3.38
Year II 2.72 c 3.06 b 2.88 bc 4.09 a 2.71 c 3.34
Mean 2.73 D 3.10 B 2.89 C 4.06 A 2.70 D 3.36
WLI Year I 0.86 b 0.77 d 0.86 b 0.80 c 0.91 a 2.41
Year II 0.86 b 0.76 d 0.85 b 0.80 c 0.91 a 2.03
Mean 0.86 B 0.76 D 0.86 B 0.80 C 0.91 A 2.23
Fruit weight
(g)
Year I 12.98 d 20.27 b 15.51 c 38.54 a 12.54 d 6.04
Year II 12.49 d 19.80 b 14.97 c 39.00 a 13.02 d 4.92
Mean 12.73 D 20.04 B 15.24 C 38.77 A 12.78 D 5.51
Fruit volume
(mm3)
Year I 12.14 d 18.76 b 14.47 c 36.09 a 11.55 d 5.91
Year II 11.66 d 18.23 b 13.99 c 36.27 a 12.03 d 5.10
Mean 11.90 D 18.50 B 14.23 C 36.18 A 11.79 D 5.53
F:S wt. Year I 2.23 bc 2.69 a 2.33 b 2.79 a 2.10 c 3.71
Year II 2.21 cd 2.65 b 2.30 c 2.82 a 2.11 d 3.61
Mean 2.22 D 2.67 B 2.32 C 2.80 A 2.11 E 3.66
F:S vol. Year I 2.49 bc 2.89 a 2.65 b 3.03 a 2.34 c 4.44
Year II 2.52 bc 2.89 a 2.66 b 3.02 a 2.36 c 3.52
Mean 2.50 D 2.89 B 2.65 C 3.03 A 2.35 E 4.01
Fruits per
bunch
Year I 14.93 a 14.10 ab 14.03 ab 14.40 a 12.87 b 2.68
Year II 14.47 a 13.60 ab 13.33 b 13.83 ab 12.20 c 3.24
Mean 14.70 A 13.85 B 13.68 B 14.12 AB 12.53 C 2.96
Days FB to M Year I 118.00 b 121.33 a 121.33 a 114.00 c 113.00 c 1.23
Year II 117.00 b 120.67 a 121.67 a 113.67 c 112.67 c 1.00
Mean 117.50 B 121.00 A 121.50 A 113.83 C 112.83 C 1.12
Yield per tree
(kg)
Year I 45.50 c 40.92 d 50.80 b 55.90 a 34.68 e 6.32
Year II 46.87 bc 44.02 c 53.07 ab 53.95 a 32.73 d 7.75
Mean 46.18 B 42.47 C 51.93 A 54.93 A 33.71 D 7.08
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by
volume; Days FB to M = Days from full bloom to maturity
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
105
Lowest seed weight was observed in TR5 (0.98 g), while lowest seed content per
fruit was recorded in TR1 (3.95 g).
Although TR1 has been observed to have the lowest number of seeds per
fruit as well as the lowest seed content per fruit, it is not so attractive due to being
the smallest one with a low flesh seed ratio. On the other hand, TR4 is an
outstanding genotype despite having the highest number of seeds as well as the
seed content per fruit, because of its biggest fruit weight (38.77 g) and the highest
flesh seed ratio among the five genotypes of Tret. Seed content of two cultivars in
Spain, ‘Saval 2’ and ‘Peluches’ has been reported to be even more i.e. 8.60 g and
11.20 g per fruit respectively (Llacer et al., 2003) but they are considered excellent
due to their larger fruits.
In Italy, ‘Selezione 2 PA’, ‘Nespolone di Trabia’ ‘Ferdinando’ and
‘Vainiglia’ have been observed to have 3.2, 3.8, 3.4 and 3.7 seeds per fruit
respectively (Insero et al., 2003). In Turkey, ‘Kanro’ ‘Baffico’ and ‘Gold Nugget’
had 2.40, 2.62 and 3.00 seeds per fruit respectively (Durgac et al., 2006). In China,
‘Niuteibaisha’ cultivar had 2.78 seeds per fruit (Feng et al., 2007). Loquat cultivars
having 1 to 2 seeds per fruit are considered ideal. ‘White loquat’ (Huang et al.,
2007) and ‘Taicheng 4’ (Xie et al., 2007) were observed to have only 2 and 1.32
seeds per fruit respectively.
4.1.4.5 Leaf characteristic
TR5 remained at the top having the highest leaf length (28.82 cm), leaf
width (8.65 cm) and leaf area (142.73 cm2). TR4 followed it with reference to leaf
106
length (28.21 cm) with non significant difference and with reference to leaf area
(126.90 cm2) with a significant difference. TR5 was followed by TR3 with
reference to leaf width (7.80 cm) with a significant difference. Lowest leaf length
was observed in TR2 (22.02 cm) which also had the lowest leaf area (100.26 cm2).
Leaf characteristics of the genotypes are given in Table 25. In the loquat cultivar,
‘Qingzhong’ leaf length and width have been reported to be 29.20 cm and 8.40 cm
respectively (Feng et al., 2007) which is comparable with the genotypes of Tret.
4.1.4.6 Inflorescence
As shown in Table 26, highest number of flowers per panicle was found in
TR1 (153.78) followed by TR4 (147.21) both being statistically at par. Lowest
number of flowers per panicle was observed in TR5 (117.02). Loquat cultivars
‘Nespolone di Trabia’, ‘Ferdinando’ and ‘Nespolone di Ficarazzi’ have been
reported to produce 108.10, 130.40 and 158.40 flowers per panicle respectively
(Insero et al., 2003). This number is comparable with that of the genotypes at Tret.
Panicle size was greatest in TR1 (17.26 cm) followed by TR5 (16.36 cm) with a
significant difference. It was lowest in TR4 (14.53 cm). TR4 took maximum time
from flowering to full bloom (36.83 days) which was significantly higher than that
of TR5 (34.17 days). Least time from flowering to full bloom was taken by TR2
(31.50 days).
107
Table 24 Seed characteristics of 5 loquat genotypes at Tret
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean TR1 3.20 d 3.17 c 3.18 D 1.25 b 1.23 b 1.24 C 4.01 d 3.89 e 3.95 D
TR2 4.25 b 4.22 b 4.23 B 1.29 b 1.29 b 1.29 B 5.50 b 5.43 b 5.46 B
TR3 4.12 c 4.15 b 4.13 C 1.13 c 1.09 c 1.11 D 4.66 c 4.53 c 4.60 C
TR4 4.82 a 4.85 a 4.83 A 2.11 a 2.10 a 2.11 A 10.16 a 10.21 a 10.19 A
TR5 4.15 bc 4.20 b 4.18 BC 0.97 d 0.99 d 0.98 E 4.04 d 4.18 d 4.11 D
CV % 1.74 1.44 1.60 3.27 2.24 2.81 3.80 2.91 3.39
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
108
Table 25 Leaf characteristics of 5 loquat genotypes at Tret
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2)
Year I Year II Mean Year I Year II Mean Year I Year II Mean TR1 23.59 b 23.80 b 23.70 B 6.90 c 6.97 b 6.94 C 104.05 cd 107.61 c 105.83 CD TR2 21.83 c 22.20 b 22.02 C 7.16 bc 7.40 b 7.28 BC 98.55 d 101.98 c 100.26 D TR3 24.26 b 23.91 b 24.08 B 7.86 ab 7.74 b 7.80 B 113.14 c 110.54 c 111.84 C TR4 28.06 a 28.35 a 28.21 A 7.09 bc 7.21 b 7.15 C 125.28 b 128.51 b 126.90 B TR5 28.28 a 29.36 a 28.82 A 8.58 a 8.72 a 8.65 A 140.53 a 144.92 a 142.73 A CV % 3.69 4.31 4.01 5.37 6.43 5.93 4.28 5.65 5.03
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
109
Table 26 Floral characteristics of 5 loquat genotypes at Tret
Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full bloom
Year I Year II Mean Year I Year II Mean Year I Year II Mean
TR1 151.73 a 155.82 a 153.78 A 16.95 a 17.56 a 17.26 A 32.67 bc 32.00 b 32.33 BC
TR2 138.12 bc 142.25 a 140.19 B 14.77 bc 15.52 cd 15.14 C 32.00 c 31.00 b 31.50 C
TR3 131.47 c 126.27 b 128.87 C 15.70 ab 16.20 bc 15.95 B 34.33 bc 33.67 ab 34.00 B
TR4 143.23 ab 151.18 a 147.21 AB 14.17 c 14.90 d 14.53 C 37.67 a 36.00 a 36.83 A
TR5 113.14 d 120.90 b 117.02 D 16.00 ab 16.72 ab 16.36 B 35.00 ab 33.33 ab 34.17 B
CV % 3.82 5.12 4.53 4.17 4.50 4.35 4.20 3.52 3.89
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
110
4.1.5 Hasan Abdal and Wah
4.1.5.1 General morphology
Genotypes HW1 and HW2 had the spreading tree habit with blunt acute
shape of leaf tip while the other three genotypes had the upright tree habit with
sharp acute shape of leaf tip. As for as shape of panicle is concerned, all the
genotypes were similar. All of them had the panicle with conical shape (Table 27).
Upright and spreading tree habit has been reported in ‘Cardona’ and ‘Peluches’
cultivars of loquat respectively in Spain. Shape of panicle in both these cultivars
has been the same i.e. conical (Llacer et al., 2003).
4.1.5.2 Fruit and seed morphology
Fruit skin colour was orange in HW1 and orange yellow in other genotypes.
Pulp colour was orange in all the genotypes except HW4 which had yellow pulp.
In Spain, orange yellow skin as well as pulp colour has been observed in a number
of loquat varieties including ‘Cardona’, ‘Algerie’, and ‘Golden Nugget’. ‘Buenet’
has orange skin and pulp colour (Llacer et al., 2003). In China, ‘Hanwuzhong’
cultivar has the same skin and pulp colour as in HW4 i.e. orange yellow and
yellow respectively (He et al., 2007). Orange pulp colour has been observed in
‘Kumquat loquat’ and ‘Sour loquat’ (Huang et al., 2007). Genotypes except HW4
had some other similarities among themselves including fruit shape (round), fruit
shape at the apex (raised) and seed colour (light brown). HW4 had obovoid fruit
shape, flat shape at the apex and brown seed colour. Fruit shape at the basal end
was obtuse in HW1, HW2 and HW3, while round in other two genotypes.
111
Table 27 General appearance of the loquat plants of 5 genotypes at
Hasan Abdal & Wah
Genotypes Tree habit Shape of leaf tip Shape of panicle
HW1 Spreading Blunt acute Conical
HW2 Spreading Blunt acute Conical
HW3 Upright Sharp acute Conical
HW4 Upright Sharp acute Conical
HW5 Upright Sharp acute Conical
Table 28 Fruit and seed morphology of 5 loquat genotypes at Hasan
Abdal & Wah
Genotypes Skin colour
Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
HW1 Orange Orange Round Obtuse Raised Light brown
Elliptic
HW2 Orange yellow
Orange Round Obtuse Raised Light brown
Elliptic
HW3 Orange yellow
Orange Round Obtuse Raised Light brown
Elliptic
HW4 Orange yellow
Yellow Obovoid Round Flat Brown Elliptic
HW5 Orange yellow
Orange Round Round Raised Light brown
Elliptic
112
Seed shape was elliptic in all the five genotypes (Table 28). Fruit shape of
HW4 resembles with that of the loquat cultivars, ‘Magdal’ and ‘Tanaka’ in Spain,
which have been reported to have obovoid shape of fruit and elliptical seeds. The
other four genotypes have round fruits and elliptical seeds as has been observed in
‘Saval-2’ (Llacer et al., 2003).
4.1.5.3 Fruit characteristics
Fruit characteristics of the genotypes are given in Table 29. Fruit length
was maximum in HW2 (3.66 cm) followed by HW4 (3.62 cm) and HW3 (3.53
cm), all the three being at par. Lowest fruit length with significant difference was
noted in HW5 (2.87 cm). Maximum fruit width was observed in HW4 (3.18 cm)
followed by HW2 (2.90 cm) with a significant difference. Fruit width was
significantly lowest in HW1 (2.53).
Width length index was highest in HW5 (0.89) followed by HW4 (0.88)
both being at par. HW1 had the lowest width length index (0.77). Fruit weight was
highest in HW4 (16.20 g) followed by HW2 (15.65 g) both being at par. Minimum
fruit weight was noted in HW5 (9.54 g) which was significantly low than the other
genotypes. Fruit volume was highest in HW4 (15.62 mm3) followed by HW2
(15.04 mm3) both being at par. It was lowest in HW5 (8.76 mm3) with a significant
difference. Flesh seed ratio by weight was highest in HW2 (2.55) followed by
HW4 (2.50) with non significant difference. Flesh seed ratio by volume was
highest in HW4 (2.97) followed by HW2 (2.94) both being at par. HW5 had the
lowest flesh seed ratio by weight (1.67) and by volume (1.90) and was significantly
113
at the bottom. Maximum number of fruits per bunch were recorded in HW1
(18.92) followed by HW5 (15.42) with a significant difference. This number was
lowest in HW4 (11.05). Time taken from full bloom to maturity was highest in
HW2 (127.83 days) which was significantly higher than HW5 (124.17 days) and
all other genotypes. This period was shortest in HW3 (121.50 days). Genotype
HW4 was at the top in terms of fruit yield per plant (50.30 kg) and was followed
by HW2 (46.41 kg) with a significant difference. Lowest yield per plant was
observed in HW5 (30.50 kg).
The highest fruit weight observed in HW4 (16.20 g) was somewhat
comparable with ‘Kumquat loquat’ (Huang et al. 2007) which had a fruit weight of
18.00 g and even greater than ‘Taishan Zhong’ cultivar which has fruit weight of
12.80 g (He et al., 2007). But such fruit size and weight is not so much admirable
because there are a number of cultivars in China, Spain, Turkey and even Pakistan
which have fruit weights many times higher than that of HW4. In Japan, ‘Satomi’
and ‘Fusakikari’ had fruit weight of 65 g and 75 g respectively (Nakai et al., 1990).
‘Hongdenglong’ in China (Jiang et al., 2001), ‘Algerie’ in Spain (Llacer et al.,
2003), ‘Nespolone di Trabia’ in Italy (Insero et al., 2003) and ‘Ottawiani’ in
Turkey (Yalcin and Paydas, 1995) were observed to have fruit weight of 63.1 g,
65.0 g. 50.40 g and 49.78 g respectively.
Genotypes of Hasan Abdal and Wah have flesh seed ratio lower than that
found in the leading loquat cultivars of the world. In Turkey, ‘Gold Nugget’,
‘Baffico’ and ‘Kanro’ in Turkey were found to have a flesh seed ratio of
114
Table 29 Fruit characteristics of 5 loquat genotypes at Hasan Abdal &
Wah
Fruit
characteristics
Year HW1 HW2 HW3 HW4 HW5 CV
%
Fruit length
(cm)
Year I 3.28 ab 3.68 a 3.53 a 3.59 a 2.88 b 6.54
Year II 3.25 b 3.65 a 3.54 ab 3.64 a 2.86 c 5.85
Mean 3.27 B 3.66 A 3.53 A 3.62 A 2.87 C 6.20
Fruit width (cm) Year I 2.55 c 2.91 ab 2.79 bc 3.15 a 2.56 c 4.62
Year II 2.50 c 2.88 b 2.79 b 3.20 a 2.53 c 4.11
Mean 2.53 C 2.90 B 2.79 B 3.18 A 2.54 C 4.37
WLI Year I 0.77 b 0.79 b 0.79 b 0.88 a 0.89 a 3.05
Year II 0.77 b 0.79 b 0.79 b 0.88 a 0.88 a 2.80
Mean 0.77 B 0.79 B 0.79 B 0.88 A 0.89 A 2..92
Fruit weight (g) Year I 13.51 b 15.86 a 14.82 ab 15.82 a 9.65 c 7.62
Year II 13.19 b 15.45 a 14.98 a 16.58 a 9.43 c 5.98
Mean 13.35 C 15.65 AB 14.90 B 16.20 A 9.54 D 6.85
Fruit volume
(mm3)
Year I 12.82 b 15.25 a 14.17 ab 15.22 a 8.87 c 7.49
Year II 12.52 c 14.84 ab 14.32 b 16.03 a 8.65 d 5.79
Mean 12.67 C 15.04 AB 14.24 B 15.62 A 8.76 D 6.70
F:S wt. Year I 2.27 b 2.56 a 2.23 b 2.50 a 1.68 c 4.27
Year II 2.23 b 2.54 a 2.25 b 2.49 a 1.66 c 3.57
Mean 2.25 B 2.55 A 2.24 B 2.50 A 1.67 C 3.94
F:S vol. Year I 2.64 b 2.94 a 2.66 b 2.98 a 1.91 c 5.34
Year II 2.61 b 2.94 a 2.62 b 2.95 a 1.89 c 4.22
Mean 2.63 B 2.94 A 2.64 B 2.97 A 1.90 C 4.82
Fruits per bunch Year I 19.10 a 14.67 bc 13.83 c 11.23 d 15.63 b 4.09
Year II 18.73 a 14.17 bc 13.30 c 10.87 d 15.20 b 5.69
Mean 18.92 A 14.42 C 13.57 C 11.05 D 15.42 B 4.93
Days FB to M Year I 123.00 b 127.67 a 121.00 b 122.33 b 123.67 b 1.14
Year II 124.33 b 128.00 a 122.00 b 123.00 b 124.67 b 0.70
Mean 123.67 B 127.83 A 121.50 C 122.67 BC 124.17 B 0.95
Yield per tree
(kg)
Year I 41.18 b 45.78 a 35.30 c 49.50 a 29.95 d 5.01
Year II 42.88 c 47.03 b 36.40 d 51.10 a 31.05 e 3.65
Mean 42.03 C 46.41 B 35.85 D 50.30 A 30.50 E 4.36
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by
volume; Days FB to M = Days from full bloom to maturity.
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
115
3.83, 4.16 and 5.42 respectively (Durgac et al., 2006). In Italy, flesh seed ratio in
‘Vainiglia’, ‘Ferdinando’ ‘Peluche’ and ‘Magdal’ have been found to be 5.4, 5.3,
5.9 and 6.5 respectively (Insero et al., 2003). In Spain ‘Cardona’, Buenet’,
‘Peluches’ and ‘Tanaka’ were observed to have high flesh seed ratios of 6.20, 7.08,
7.48 and 5.38 respectively (Llacer et al., 2003).
Fruit yield per plant in the genotypes of Hasan Abdal and Wah is higher
than that of ‘Kanro’ (Karadeniz, 2003), which produced 24.5 kg fruit per plant. But
the leading loquat cultivars have been reported to give much better yields in the
other countries. Yield per plant in ‘M. Marie’, ‘Champagne de Grasse’ (Karadeniz,
2003) in Turkey and ‘Algarie’ (Hermeso and Farre, 2003) in Spain has been
reported to be 69 kg, 70 kg and 74 kg respectively.
4.1.5.4 Seed characteristics
Significant differences were observed among the genotypes with respect to
the seed characteristics studied (Table 30). Highest number of seeds per fruit was
observed in HW3 (4.69) followed by HW4 (3.85) with a significant difference.
This number was lowest in HW1 (2.99). All the genotypes were significantly
different with respect to seed weight. Seed weight was highest in HW1 (1.38 g)
followed by HW2 (1.28 g) while minimum in HW3 (0.98 g).
Genotypes HW4, HW3 and HW2 remained at top with reference to seed
content per fruit (4.64 g. 4.61 g and 4.40 g respectively). These genotypes
remained at par with each other while significantly different from the other
genotypes with reference to seed content per fruit. The lowest seed content per
116
fruit was observed in HW5 (3.58 g) which was significantly low than that of all
other genotypes. In Italy, ‘Algarie’ variety has 4.10 seeds per fruit and average
seed content per fruit is much higher (7.90 g) than the above genotypes. But the
fruit weight of Algarie (57.70 g) is more than three times the weight of HW4
(Insero et al., 2003). ‘Crisanto Amadeo’ and ‘Buenet’ in Spain had 3.60 and 2.50
seeds per fruit respectively (Llacer et al., 2003). In China, ‘Taicheng 4’ (Xie et al.,
2007) and ‘White loquat’ (Huang et al., 2007) were reported to have only 1.32 and
2 seeds per fruit respectively.
4.1.5.5 Leaf characteristic
Leaf characteristics of the genotypes are given in Table 31. Leaf length was
highest in HW1 (26.01 cm) followed by HW5 (25.73 cm) both being at par. All
other genotypes were significantly different from these genotypes as well as from
one another. Lowest leaf length was noted in HW3 (22.02 cm). Maximum leaf
width was observed in HW5 (8.29 cm) which was followed by HW1 (7.24 cm)
with a significant difference. Minimum leaf width was recorded in HW4 (6.76 cm).
HW5 was also at the top in terms of leaf area (138.90 cm2) followed by HW1
(124.19 cm2) with a significant difference. The other three genotypes were
significantly different from HW1 and HW5 while at par among one another, HW4
having the lowest leaf area (100.57 cm2). Loquat cultivar, ‘Jidanbai’ in China has a
leaf length of 25.50 cm and leaf width of 7.80 cm, while the leaf length and leaf
width in case of ‘Ruantiaobaisha’ was 21.1 cm and 7.1 cm respectively (Feng et
al., 2007). Leaf size of these cultivars is comparable with that of the genotypes at
Hasan Abdal and Wah garden.
117
4.1.5.6 Inflorescence
Number of flowers per panicle was highest in HW1 (174.02) followed by
HW5 (171.96) and HW3 (168.57) with non significant difference. This number
was significantly low in the other two genotypes, HW2 having the lowest number
of flowers per panicle (150.54). In Italy, loquat cultivars Vainiglia’ and ‘Nespolone
di Ficarazzi’ have been observed to give 176.20 and 158.40 flowers per panicle
respectively (Insero et al., 2003). This number is comparable with that of the
genotypes of Hasan Abdal & Wah garden. All the genotypes significantly differed
in terms of panicle size. It was maximum in HW1 (22.60 cm) followed by HW4
(21.70 cm) while minimum in HW5 (18.10 cm). HW1 took the maximum time
from flowering to full bloom (42.50 days). HW2 and HW4 were at par with HW1
having this time as 42.00 days and 41.17 days respectively. HW5 took the
minimum time from flowering to full bloom (37.83 days) and was significantly
different from all other genotypes (Table 32).
4.1.6 Hari Pur
4.1.6.1 General morphology
All the three genotypes had the spreading tree habit and blunt acute shape
of leaf tip. Shape of panicle in all the three genotypes was also the same i.e. conical
(Table 33). Tree habit and shape of panicle in these genotypes has a resemblance
with the loquat cultivar, ‘Peluches’, in Spain, which has a spreading tree habit and
conical shape of panicle (Llacer et al., 2003).
118
Table 30 Seed characteristics of 5 loquat genotypes at Hasan Abdal & Wah
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
HW1 2.97 c 3.02 d 2.99 D 1.39 a 1.36 a 1.38 A 4.13 ab 4.09 c 4.11 B
HW2 3.42 bc 3.45 c 3.43 C 1.30 b 1.26 b 1.28 B 4.45 a 4.36 bc 4.40 A
HW3 4.67 a 4.72 a 4.69 A 0.98 d 0.98 d 0.98 E 4.59 a 4.62 ab 4.61 A
HW4 3.83 b 3.87 b 3.85 B 1.18 c 1.23 b 1.21 C 4.52 a 4.75 a 4.64 A
HW5 3.17 c 3.23 cd 3.20 CD 1.14 c 1.10 c 1.12 D 3.60 b 3.55 d 3.58 C
CV % 6.64 5.55 6.11 3.66 2.91 3.31 6.68 4.07 5.53
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
119
Table 31 Leaf characteristics of 5 loquat genotypes at Hasan Abdal & Wah
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
HW1 26.31 a 25.72 a 26.01 A 7.29 b 7.18 b 7.24 B 125.89 b 122.49 b 124.19 B
HW2 24.38 b 24.92 ab 24.65 B 7.01 b 7.11 b 7.06 BC 104.52 c 107.27 c 105.90 C
HW3 21.65 c 22.38 c 22.02 D 7.01 b 7.08 b 7.05 BC 101.28 c 103.75 c 102.52 C
HW4 22.77 c 23.54 bc 23.16 C 6.80 b 6.71 b 6.76 C 99.40 c 101.74 c 100.57 C
HW5 26.01 a 25.44 a 25.73 A 8.33 a 8.25 a 8.29 A 140.37 a 137.44 a 138.90 A
CV % 3.22 3.69 3.46 4.09 4.05 4.07 6.54 6.52 6.53
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
120
Table 32 Floral characteristics of 5 loquat genotypes at Hasan Abdal & Wah
Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full
bloom Year I Year II Mean Year I Year II Mean Year I Year II Mean
HW1 171.22 a 176.82 a 174.02 A
22.45 a
22.74 a 22.60 A
43.00 a 42.00 a 42.50 A
HW2 148.50 c 152.58 c 150.54 B
21.32 b
21.66 ab
21.49 B
42.33 a 41.67 ab 42.00 AB
HW3 165.30 ab
171.83 ab
168.57 A
20.18 c
20.59 b 20.39 C
41.33 a 39.67 bc 40.50 B
HW4 152.27 bc
155.12 bc
153.69 B
21.59 b
21.80 a 21.70 B
42.00 a 40.33 ab 41.17 AB
HW5 168.75 a 175.17 a 171.96 A
17.97 d
18.22 c 18.10 D
38.33 b 37.33 c 37.83 C
CV % 4.70 5.36 5.05 2.11 2.80 2.48 3.33 2.13 2.81
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
121
4.1.6.2 Fruit and seed morphology
Fruit skin colour as well as pulp colour was orange yellow in HP1 and HP2
while yellowish white in HP3. Fruit shape was also the same i.e. obovoid in HP1
and HP2 whereas it was round in HP3. Fruit shape at the basal end was obtuse in
HP1 and round in HP2 and HP3. Fruit shape at the apex was raised in HP1 and
HP2 while it was flat in HP3. HP1 and HP2 had the seeds with light brown colour
and elliptic shape while HP3 had those with brown colour and round shape (Table
34).
Orange yellow skin as well as pulp colour has also been observed in a
number of loquat varieties including ‘Cardona’, ‘Algerie’ and ‘Golden Nugget’
(Llacer et al., 2003). ‘Qingbian’ in China has a yellowish white skin colour and
white pulp colour (He et al., 2007). Obovoid fruit shape has been observed in
‘Magdal’, ‘Golden Nugget’ and ‘Tanaka’ (Llacer et al., 2003), while ‘Donghuzao’
(Zhao et al., 2001), ‘Ningbai’, ‘Qingzhong’ and ‘Baili’ (Feng et al., 2007) were
found to have round shape fruit. In Spain, elliptical seed shape has been reported in
‘Magdal’, ‘Algerie’ and ‘Cardona’, while round in ‘Golden Nugget’ and ‘Crisanto
Amadeo’ (Llacer et al., 2003).
4.1.6.3 Fruit characteristics
Fruit characteristics of the genotypes are given in Table 35. Significant
differences were observed among the different genotypes. Fruit length was highest
in HP3 (3.52 cm) followed by HP2 (3.35 cm) and lowest in HP1 (3.15 cm)
122
Table 33 General appearance of the loquat plants of 3 genotypes at Hari
Pur
Genotypes Tree habit Shape of leaf tip Shape of panicle
HP1 Spreading Blunt acute Conical
HP2 Spreading Blunt acute Conical
HP3 Spreading Blunt acute Conical
Table 34 Fruit and seed morphology of 3 loquat genotypes at Hari Pur
Genotypes Skin colour
Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
HP1 Orange yellow
Orange yellow
Obovoid Obtuse Raised Light brown
Elliptic
HP2 Orange yellow
Orange yellow
Obovoid Round Raised Light brown
Elliptic
HP3 Yellowish white
Yellowish white
Round Round Flat Brown Round
123
all being significantly different from one another. Fruit width was highest in HP1
(3.35 cm) followed by HP3 (3.32 cm) with a non significant difference. It was
significantly low in HP2 (3.08 cm). Width length index was maximum in HP1
(1.06) followed by HP3 (0.95) and minimum in HP2 (0.92) all having the
significant difference.
Fruit weight was significantly highest in HP3 (24.36 g) while HP1 and HP2
remained at par having fruit weight of 20.41 g and 19.54 g respectively. Similar
was the case with fruit volume which was significantly highest in HP3 (23.31
mm3). The other two genotypes i.e. HP1 and HP2 had had a fruit volume 19.47
mm3 and 18.59 mm3 respectively and remained at par with one another. Flesh seed
ratio by weight as well as by volume was highest in HP1 (2.66 and 3.06
respectively). HP3 was at par with HP1 having these values as 2.60 and 2.99
respectively). HP2 remained significantly at the bottom with reference to flesh
seed ratio by weight (2.41) as well as by volume (2.83). Number of fruits per
bunch was highest in HP2 (20.62) followed by HP1 (15.70) and was lowest in HP3
(15.15) all the three being significantly different from one another. HP3 took
maximum time from full bloom to maturity (127.00 days) which was significantly
higher than that of HP2 (122.67). HP1 took the least time from full bloom to
maturity (115.67 days) with a significant difference from the other two genotypes.
Yield per tree was highest in HP3 (55.57 kg) followed by HP1 (46.06 kg) while
lowest in HP2 (41.53 kg) all being significantly different.
The highest fruit weight observed in HP3 (24.36 g) is almost the same as in
‘Libai’ and ‘Niuteibaisha’ having fruit weight 22.80 g and 24.70 g respectively
124
(Feng et al., 2007). But there are a number of varieties growing in other loquat
growing countries which produce much larger fruit. ‘Zaozhong 6’ (Zheng, 2001),
‘Hanwuzhong’ and ‘Mojia No. 1’ (He et al., 2007) in China have fruit weight of
52.7 g, 30.97 g and 53.20 g respectively. ‘Tanaka’ and ‘Algerie’, two popular
cultivars in Spain have fruit weight of 60.60 g and 65.00 g respectively (Llacer et
al., 2003). ‘Tanaka’ cultivar produces fruits with average weight of 70 g in Japan
(Durgac et al., 2006). Flesh seed ratio in HP1 (2.66) and other loquat genotypes of
Haripur is low as compared with a lot of cultivars growing in China, Spain and
Turkey. In Italy, flesh seed ratio in ‘Nespolone di Ficarazzi’, ‘Vainiglia’,
‘Ferdinando’ and ‘Magdal’ have been found to be 5.7, 5.4, 5.3 and 6.5
respectively (Insero et al., 2003). Similarly, ‘Dr Trabut’,‘Baffico’ and ‘Kanro’ in
Turkey have been found to have a flesh seed ratio of 3.79, 4.16 and 5.42
respectively (Durgac et al., 2006).
The lowest yield of 41.53 kg per plant obtained from HP2 is very high as
compared with that of ‘Kanro’ (Karadeniz, 2003) which had a yield of 24.5 kg per
plant. On the other hand, the highest yield given by the genotype HP3 (55.57 kg
per plant) is smaller than that of ‘Champagne de Grasse’ (Karadeniz, 2003), ‘Gold
Nugget’ and ‘Algerie’ (Hermoso and Farre, 2003) having yields of 69 kg, 72 kg
and 74 kg per plant respectively.
4.1.6.4 Seed characteristics
Number of seeds per fruit was highest in HP1 (3.63) followed by HP2
125
Table35 Fruit characteristics of 3 loquat genotypes at Hari Pur
Fruit characteristics Year HP1 HP2 HP3 CV %
Fruit length (cm) Year I 3.16 c 3.36 b 3.50 a 0.84
Year II 3.14 c 3.35 b 3.53 a 0.87
Mean 3.15 C 3.35 B 3.52 A 0.85
Fruit width (cm) Year I 3.37 a 3.10 b 3.31 a 1.12
Year II 3.33 a 3.06 b 3.33 a 1.21
Mean 3.35 A 3.08 B 3.32 A 1.17
WLI Year I 1.07 a 0.92 c 0.94 b 0.54
Year II 1.06 a 0.91 c 0.95 b 0.69
Mean 1.06 A 0.92 C 0.95 B 0.62
Fruit weight (g) Year I 20.72 ab 19.84 b 24.10 a 5.52
Year II 20.09 b 19.24 b 24.62 a 4.24
Mean 20.41 B 19.54 B 24.36 A 4.93
Fruit volume
(mm3)
Year I 19.81 b 18.88 b 23.11 a 6.92
Year II 19.12 b 18.29 b 23.52 a 3.67
Mean 19.47 B 18.59 B 23.31 A 5.56
F:S wt. Year I 2.66 a 2.43 b 2.59 a 2.18
Year II 2.65 a 2.39 b 2.60 a 2.29
Mean 2.66 A 2.41 B 2.60 A 2.24
F:S vol. Year I 3.06 a 2.84 b 3.00 a 1.92
Year II 3.05 a 2.82 b 2.97 a 2.00
Mean 3.06 A 2.83 B 2.99 A 1.96
Fruits per bunch Year I 15.43 b 20.40 a 18.80 c 6.02
Year II 15.97 b 20.83 a 15.50 c 5.52
Mean 15.70 B 20.62 A 15.15 C 5.77
Days FB to M Year I 114.67 c 121.33 b 126.00 a 1.59
Year II 116.67 b 124.00 a 128.00 a 1.90
Mean 115.67 C 122.67 B 127.00 A 1.75
Yield per tree (kg) Year I 44,63 b 40.65 c 54.05 a 2.28
Year II 47.48 b 42.40 c 57.08 a 4.05
Mean 46.06 B 41.53 C 55.57 A 3.33
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by volume; Days FB to M = Days from full bloom to maturity
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
126
(3.48) while lowest in HP3 (3.15), all being significantly different from one
another. HP3 had the highest seed weight (2.15 g) followed by HP2 (1.65 g) with a
significant difference. Significantly lowest seed weight was observed in HP1 (1.53
g). Seed content per fruit was highest in HP3 (6.77 g) followed by HP2 (5.73 g)
with a significant difference. HP1 had the lowest seed content per fruit (5.58 g) and
remained at par with HP2 (Table 36).
In Spain, ‘Algerie’, ‘Buenet’ and ‘Golden Nugget’ exhibited 2.3, 2.5 and
3.2 seeds respectively, while seed content in these cultivars was 7.3 g, 7.2 g and
8.1 g respectively (Llacer et al., 2003). But this much seed content is not so high
while keeping in view much bigger fruits (65 g, 58.2 g and 54.6 g respectively),
yielding high recovery of pulp in case of these cultivars.
4.1.6.5 Leaf characteristic
Leaf characteristics of the genotypes are given in Table 37. Leaf length was
maximum in HP2 (19.90 cm) followed by HP1 (19.84 cm) both being at par with
each other. It was significantly low in HP3 (18.25 cm). Genotypes had non
significant differences with reference to leaf width as well leaf area. Leaf size of
these genotypes is comparable with that of ‘Mojia No. 1’ in China which had a leaf
length of 19.80 cm and leaf width of 5.20 cm (He et al., 2007). Another loquat
cultivar, ‘Ruantiaobaisha’ had 21.1 cm and 7.1 cm leaf length and width
respectively (Feng et al., 2007).
127
4.1.6.6 Inflorescence
The three genotypes had non significant differences with reference to the
number of flowers per panicle which ranged from 154.46 to 170.27, which is
comparable with ‘Cardona’, giving 168 flowers per panicle, and ‘Italiano 1’,
giving 160 flowers per panicle in Spain (Llacer et al., 2003). In Italy, loquat
cultivars ‘Vainiglia’ and ‘Nespolone di Ficarazzi’ have been reported to produce
176.20 and 158.40 flowers per panicle respectively (Insero et al., 2003). Maximum
panicle size was observed in HP2 (24.42 cm) followed by HP1 (21.87 cm) while it
was minimum in HP3 (21.04 cm), all having significant differences among one
another. No significant differences were noted with reference to days from
flowering to full bloom (Table 38).
4.1.7 Mardan
4.1.7.1 General morphology
All the three genotypes had the spreading tree habit and blunt acute shape
of leaf tip. Shape of panicle was Cylindrical in MN3 while conical in the other two
genotypes (Table 39). In Spain, ‘Peluches’ cultivar of loquat has a spreading tree
habit and shape of panicle in this cultivar is conicle (Llacer et al., 2003).
4.1.7.2 Fruit and seed morphology
Fruit skin colour was orange yellow in all three genotypes. Pulp colour in
MN2 was orange yellow, while orange in the other two genotypes. Fruit shape was
round in MN3 while obovoid in MN1 and MN2. Fruit shape at the basal end
128
Table 36 Seed characteristics of 3 loquat genotypes at Hari Pur
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
HP1 3.62 a 3.65 a 3.63 A 1.56 b 1.51 b 1.53 C 5.65 b 5.50 b 5.58 B
HP2 3.47 a 3.48 b 3.48 B 1.67 b 1.63 b 1.65 B 5.79 b 5.67 b 5.73 B
HP3 3.13 b 3.17 c 3.15 C 2.14 a 2.16 a 2.15 A 6.71 a 6.84 a 6.77 A
CV % 2.13 1.19 1.72 5.17 2.91 4.21 6.20 3.11 4.91
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
129
Table 37 Leaf characteristics of 3 loquat genotypes at Hari Pur
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
HP1 20,02 a 19.66 a 19.84 A 5.65 ns 5.88 ns 5.77 ns 75.67 ns 78.07 ns 76.87 ns
HP2 20,01 a 19.78 a 19.90 A 5.68 ns 5.80 ns 5.74 ns 75.99 ns 78.14 ns 77.07 ns
HP3 18.51 b 17.98 b 18.25 B 6.30 ns 6.14 ns 6.22 ns 78.19 ns 75.89 ns 77.04 ns
CV % 2.73 2.38 2.56 5.33 6.00 5.68 4.94 5.04 4.99
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
130
Table 38 Floral characteristics of 3 loquat genotypes at Hari Pur
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full bloom
Year I Year II Mean Year I Year II Mean Year I Year II Mean
HP1 157.49 ns 164.86 ns 161.18 ns 21.57 b 22.17 b 21.87 B 44.33 ns 43.00 ns 43.67 ns
HP2 167.09 ns 173.45 ns 170.27 ns 24.30 a 24.54 a 24.42 A 41.67 ns 40.00 ns 40.83 ns
HP3 150.09 ns 158.83 ns 154.46 ns 20.82 c 21.26 c 21.04 C 41.33 ns 40.00 ns 40.67 ns
CV % 7.28 4.65 6.05 1.92 2.06 2.00 4.48 5.63 5.07
131
Table 39 General appearance of the loquat plants of 3 genotypes at
Mardan
Genotypes Tree habit Shape of leaf tip Shape of panicle
MN1 Spreading Blunt acute Conical
MN2 Spreading Blunt acute Conical
MN3 Spreading Blunt acute Cylindrical
Table 40 Fruit and seed morphology of 3 loquat genotypes at Mardan
Genotype codes
Skin colour
Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
MN1 Orange yellow
Orange Obovoid Obtuse Flat Light brown
Round
MN2 Orange yellow
Orange yellow
Obovoid Obtuse Raised Light brown
Elliptic
MN3 Orange yellow
Orange Round Rounded Depressed Brown Elliptic
132
was round in MN3 whereas obtuse in the other two genotypes. Fruit shape at the
apex was flat in MN1, raised in MN2 and depressed in MN3. MN1 and MN2 had
the seeds with light brown colour, while seed colour in MN3 was brown. Shape of
seeds was round in MN1, while elliptic in the other two genotypes (Table 40).
Orange yellow skin as well as pulp colour has also been reported in
‘yangmeizhou 4’ (Wu, 2001), ‘Algerie’, ‘Cardona’ and ‘Tanaka’ while ‘Italiano 1’
and ‘Buenet’ have been observed to have orange colour in skin as well as in pulp
(Llacer et al., 2003). Obovoid fruit shape has been noted in ‘Magdal’, ‘Golden
Nugget’ and ‘Tanaka’ (Llacer et al., 2003), while ‘Donghuzao’ (Zhao et al., 2001),
‘Ningbai’, ‘Qingzhong’ and ‘Baili’ (Feng et al., 2007) were found to have round
shape fruit. Elliptical seed shape has been reported in ‘Algerie’, ‘Magdal’ and
‘Cardona’, while round in ‘Crisanto Amadeo’ and ‘Golden Nugget’ (Llacer et al.,
2003).
4.1.7.3 Fruit characteristics
Significant differences were observed with reference to fruit characteristics
among the three genotypes (Table 41). Maximum fruit length was observed in
MN2 (4.63 cm) followed by MN1 (3.99 cm). It was lowest in MN3 (3.87 cm).
Fruit width was highest in MN2 (3.52 cm) followed by MN3 (3.22 cm) with a
significant difference. MN1 had the lowest fruit width (3.19 cm) and remained at
par with MN3. Width length index was highest in MN3 (0.83) followed by MN1
(0.80) and lowest in MN2 (0.76) all being significantly different from one another.
Highest fruit weight was recorded in MN2 (43.78 g) followed by MN3 (36.66 g)
133
while lowest in MN1 (23.05 g) all having significant differences. Similarly, highest
fruit volume was observed in MN2 (42.13 mm3) followed by MN3 (35.17 mm3)
while lowest in MN1 (22.03 mm3). Flesh seed ratio by weight was highest in MN2
(2.77) followed with significant difference by MN3 (2.35) which was afterward
significantly higher than MN1 (2.12). The same order existed in flesh seed ratio by
volume which was highest in MN2 (3.28) and lowest in MN1 (2.46). Number of
fruits per bunch was highest in MN1 (14.83) followed by MN2 (11.62) and lowest
in MN3 (8.90) showing significant differences. There were non significant
differences regarding days from full bloom to maturity.
Yield per tree was highest in MN3 (77.44 kg) followed by MN2 (69.23 kg)
and lowest in MN1 (59.35 kg), all being significantly different from one another.
These yields are comparable with the fruit yields of word known loquat cultivars.
MN3 has even better yield than that of ‘Champagne de Grasse’ in Turkey
(Karadeniz, 2003), ‘Gold Nugget’ and ‘Algarie’ in Spain (Hermoso and Farre,
2003), which had a yield of 70 kg, 72 kg and 74 kg per tree respectively.
Highest fruit weight observed in MN2 (43.78 g) is almost the same as that
in ‘Ningbai 3’, having a weight of 43.80 g (Feng et al., 2007). It is approximately
double the size of ‘Maomu’ (Luo, 2005), which had a fruit weight of 21.7 g. Fruit
weight of MN2 is also greater than the thirteen loquat types (b1 to b13) described
by Polat (2007) in Turkey, wherein fruit weight ranged from 20.50 g to 39.21 g.
But it is comparatively low from that of some top class varieties of China and
134
Table 41 Fruit characteristics of 3 loquat genotypes at Mardan
Fruit
characteristics
Year MN1 MN2 MN3 CV %
Fruit length (cm) Year I 4.01 b 4.63 a 3.89 b 1.49
Year II 3.97 b 4.64 a 3.86 c 1.16
Mean 3.99 B 4.63 A 3.87 C 1.34
Fruit width (cm) Year I 3.19 b 3.54 a 3.24 b 1.99
Year II 3.18 b 3.50 a 3.20 b 1.74
Mean 3.19 B 3.52 A 3.22 B 1.87
WLI Year I 0.80 b 0.77 c 0,83 a 1.22
Year II 0.80 b 0.75 c 0.83 a 0.84
Mean 0.80 B 0.76 C 0.83 A 1.05
Fruit weight (g) Year I 23.23 c 44.07 a 36.97 b 4.34
Year II 22.87 c 43.50 a 36.35 b 3.74
Mean 23.05 C 43.78 A 36.66 B 4.05
Fruit volume
(mm3)
Year I 22.14 c 42.29 a 35.41 b 4.28
Year II 21.93 c 41.97 a 34.93 b 3.72
Mean 22.03 C 42.13 A 35.17 B 4.01
F:S wt. Year I 2.12 c 2.78 a 2.35 b 2.40
Year II 2.12 c 2.75 a 2.36 b 2.20
Mean 2.12 C 2.77 A 2.35 B 2.30
F:S vol. Year I 2.42 c 3.29 a 2.77 b 3.98
Year II 2.49 c 3.27 a 2.83 b 1.97
Mean 2.46 C 3.28 A 2.80 B 3.13
Fruits per bunch Year I 14.67 a 11.50 b 8.77 c 7.61
Year II 15.00 a 11.73 b 9.03 c 5.69
Mean 14.83 A 11.62 B 8.90 C 6.69
Days FB to M Year I 127.3 ns 126.0 ns 126.7 ns 1.02
Year II 127.0 ns 125.7 ns 126.7 ns 0.62
Mean 127.2 ns 125.8 ns 126.2 ns 0.84
Yield per tree
(kg)
Year I 59.95 c 70.03 b 78.50 a 3.56
Year II 58.75 c 68.43 b 76.38 a 1.79
Mean 59.35 C 69.23 B 77.44 A 2.84
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by
volume; Days FB to M = Days from full bloom to maturity
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
135
Spain, where ‘Hongdenglong’ (Jiang et al., 2001), ‘Donghuzao’ (Zhao et al.,
2001), ‘Algarie’ and ‘Crisanto Amadeo’ have fruit weight of 63.1 g, 59.2 g, 65 g
and 68.7 g respectively.
Highest flesh seed ratio by weight observed in MN2 (2.77) is reasonably
satisfactory, but it is comparatively low than that of the leading loquat cultivars of
the world. Flesh seed ratio was much higher in ‘Selezione 2 PA’ (4.8),
‘Ferdinando’ (5.3), ‘Algerie’ (6.2) as observed in Italy (Insero et al., 2003), ‘Gold
Nugget’ (3.83) ‘Kanro’, (5.42) and ‘Bafico’ (4.16) as noted in Turkey (Durgac et
al., 2006).
4.1.7.4 Seed characteristics
Significant differences were observed among the genotypes with respect to
all the seed characteristics studied (Table 42). MN3 had the highest number of
seeds per fruit (4.98) followed by MN2 (4.67), while this number was lowest in
MN1 (3.62). Maximum seed weight was observed in MN2 (2.49 g) followed by
MN3 (2.20 g) with significant difference. Minimum seed weight with significant
difference was noted in MN1 (2.01 g). Seed content per fruit was highest in MN2
(11.62 g) followed by MN3 (10.94 g) while lowest in MN1 (7.39 g) all having
significant differences.
Though lowest number of seeds per fruit, seed weight as well as seed
content per fruit has been observed in MN1, it is not an outstanding genotype due
to being the smallest one with the lowest flesh seed ratio. On the other hand, MN2
is an excellent genotype despite having the highest number of seeds, seed weight
136
as well as the seed content per fruit because of its biggest fruit (43.78 g) and the
highest flesh seed ratio among the three genotypes of Mardan. ‘Peluches’, a
cultivars in Spain, has been reported to have high seed content of 11.20 g per fruit
(Llacer et al., 2003), which is almost the same as that in MN2 (11.62 g per fruit)
but is admirable due to its very large fruits weighing 95.00 g on an average.
Lowest number of seeds per fruit (3.68) observed in MN1, is slightly greater than
that of ‘Niuteibaisha’ cultivar in China having 2.78 seeds per fruit (Feng et al.,
2007). In China, some cultivars with 2 or less seeds per fruit have also been
reported. ‘White loquat’ (Huang et al., 2007) and ‘Taicheng 4’ (Xie et al., 2007)
were observed to have only 2 and 1.32 seeds per fruit respectively
4.1.7.5 Leaf characteristic
Highest leaf length was noted in MN3 (25.84 cm) followed by MN2 (25.19
cm) while lowest in MN1 (23.94 cm), all the three having significant differences
among one another. Leaf width was maximum in MN3 (8.14 cm) followed by
MN1 (7.93 cm) with non significant difference, while significantly minimum in
MN2 (7.02 cm). The maximum leaf area was exhibited by MN3 (145.8 cm2) which
was followed with a significant difference by MN1 (134.9 cm2). It was
significantly low (119.4 cm2) in MN2 (Table 43). Leaf size of these genotypes is
comparable with that of ‘Ningbai 1’ in China which had a leaf length of 25.50 cm
and leaf width of 8.00 cm. Leaf length and width in ‘Jindanbai’ was reported to be
25.50 cm and 7.8 cm respectively (Feng et al., 2007). ‘Hanwuzhong’ had leaf
length of 26.50 cm and leaf width of 9.00 cm (He et al., 2007).
137
4.1.7.6 Inflorescence
Significant differences were observed among the genotypes with respect to
the leaf characteristics studied (Table 44). Number of flowers per panicle was
highest in MN2 (163.4) followed by MN1 (154.4) while minimum in MN3
(136.4). Number of flowers per panicle in these genotypes is comparable with the
Spanish cultivars, ‘Ferdinando’, ‘Nespolone di Ficarazzi’ and ‘Golden Nugget’,
which produced 130.40, 158.40 and 165.30 flowers per panicle respectively
(Llacer et al., 2003). MN1 remained at the top with reference to panicle size (21.43
cm) and was followed by MN2 (19.36 cm). MN3 had the lowest size of the panicle
(18.58 cm) with a significant difference. MN1 took the maximum time from
flowering to full bloom (46.33 days). MN2 took comparatively less time (44.00
days) with non significant difference. Least time from flowering to full bloom was
taken by MN3 (41.50 days) which was significantly low as compared with that of
the other two genotypes.
4.1.8 Takht Bhai
4.1.8.1 General morphology
Genotypes TB3 and TB11 had upright tree habit; TB8, TB12 and TB15 had
spreading tree habit, while all other genotypes had semi upright tree habit. Shape
of leaf tip was sharp acute in TB6, whereas blunt acute in all other genotypes.
Shape of panicle was truncate conical in TB1, TB4 and TB10; it was cylindrical in
TB3, TB6, TB7 and TB15 while conical in all other genotypes (Table 45).
138
Table 42 Seed characteristics of 3 loquat genotypes at Mardan
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
MN1 3.65 c 3.72 c 3.68 C 2.04 c 1.97 c 2.01 C 7.44 b 7.34 c 7.39 C
MN2 4.65 b 4.68 b 4.67 B 2.50 a 2.48 a 2.49 A 11.64 a 11.59 a 11.62 A
MN3 4.97 a 4.98 a 4.98 A 2.22 b 2.17 b 2.20 B 11.04 a 10.83 b 10.94 B
CV % 2.43 2.53 2.48 1.02 1.67 1.38 3.04 2.92 2.98
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
139
Table 43 Leaf characteristics of 3 loquat genotypes at Mardan
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
MN1 23.57 b 24.32 b 23.94 C 7.87 a 7.99 a 7.93 A 133.5 a 136.4 a 134.9 B
MN2 25.40 a 24.99 a 25.19 B 7.07 b 6.96 b 7.02 B 120.8 b 117.9 b 119.4 C
MN3 25.96 a 25.72 a 25.84 A 8.09 a 8.20 a 8.14 A 144.9 a 146.8 a 145.8 A
CV % 2.66 1.07 2.02 2.97 2.79 2.88 3.87 5.26 4.62
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
140
Table 44 Floral characteristics of 3 loquat genotypes at Mardan
Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full bloom
Year I Year II Mean Year I Year II Mean Year I Year II Mean
MN1 152.9 b 155.9 a 154.4 B 21.36 a 21.51 a 21.43 A 46.67 a 46.00 a 46.33 A
MN2 167.1 a 156.6 a 163.4 A 19.27 b 19.44 b 19.36 B 44.67 ab 43.33 b 44.00 B
MN3 132.3 c 140.5 b 136.4 C 18.46 b 18.69 b 18.58 C 42.00 b 41.00 b 41.50 C
CV % 4.75 4.30 4.53 2.60 2.79 2.70 2.86 2.60 2.73
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
141
In literature, upright, semi upright and spreading tree habit has been
reported in the loquat culticars ‘Cardona’, ‘Italiano 1’ and ‘Peluches’ respectively,
all having the same shape of panicles i.e. conical (Llacer et al., 2003).
4.1.8.2 Fruit and seed morphology
Fruit skin colour was yellow in TB5 and TB11, orange in TB13, yellowish
white in TB8 and TB15 while orange yellow in all other genotypes. Pulp colour
was yellowish white in TB6, orange yellow in TB1, TB3, TB8, TB9 and TB15
while orange in all other genotypes. Orange yellow skin as well as pulp colour has
also been reported in a number of loquat varieties including ‘Cardona’, ‘Algerie’
and ‘Golden Nugget’ (Llacer et al., 2003). ‘Qingbian’ in China has a yellowish
white skin colour and white pulp colour (He et al., 2007). Fruit shape was oblong
in TB1 and TB10, round in TB3 and TB4 but obovoid in all other genotypes. Fruit
shape at the basal end was acute in TB1 and TB2, round in TB4, TB5, TB8 and
TB15 whereas obtuse in the remaining genotypes. Fruit shape at the apex was
raised in TB1, TB2, TB9, TB10, TB11 and TB14, depressed in TB3 and TB7
while flat in all other genotypes. Seed colour was brown in TB1 and dark brown in
TB2. All other genotypes had the seeds with light brown colour. Seed shape was
round in TB4 and TB5, while elliptical in all the remaining genotypes (Table 46).
Obovoid fruit shape has been observed in ‘Magdal’, ‘Golden Nugget’ and
‘Tanaka’ (Llacer et al., 2003), while ‘Donghuzao’ (Zhao et al., 2001), ‘Ningbai’,
‘Qingzhong’ and ‘Baili’ (Feng et al., 2007) were found to have round shaped fruit.
142
Table 45 General appearance of the loquat plants of 15 genotypes at Takht
Bhai
Genotype codes Tree habit Shape of leaf tip Shape of panicle
TB1 Semi upright Blunt acute Truncate conical
TB2 Semi upright Blunt acute Conical
TB3 Upright Blunt acute Cylindrical
TB4 Semi upright Blunt acute Truncate conical
TB5 Semi upright Blunt acute Conical
TB6 Semi upright Sharp acute Cylindrical
TB7 Semi upright Blunt acute Cylindrical
TB8 Spreading Blunt acute Conical
TB9 Semi upright Blunt acute Conical
TB10 Semi upright Blunt acute Truncate conical
TB11 Upright Blunt acute Conical
TB12 Spreading Blunt acute Conical
TB13 Semi upright Blunt acute Conical
TB14 Semi upright Blunt acute Conical
TB15 Spreading Blunt acute Cylindrical
143
Table 46 Fruit and seed morphology of 15 loquat genotypes at Takht Bhai
Genotype codes
Skin colour
Pulp colour
Fruit shape
Fruit shape at the basal end
Fruit shape at the apex
Seed colour
Seed shape
TB1 Orange yellow
Orange yellow
Oblong Acute Raised Brown Elliptic
TB2 Orange yellow
Orange Obovoid Acute Raised Dark brown
Elliptic
TB3 Orange yellow
Orange yellow
Round Obtuse Depressed Light brown
Elliptic
TB4 Orange yellow
Orange Round Round Flat Light brown
Round
TB5 Yellow Orange Obovoid Round Flat Lt. brown Round TB6 Orange
yellow Yellowish white
Obovoid Obtuse Flat Light brown
Elliptic
TB7 Orange yellow
Orange Obovoid Obtuse Depressed Light brown
Elliptic
TB8 Yellowish white
Orange yellow
Obovoid Round Flat Light brown
Elliptic
TB9 Orange yellow
Orange yellow
Obovoid Obtuse Raised Light brown
Elliptic
TB10 Orange yellow
Orange Oblong Obtuse Raised Light brown
Elliptic
TB11 Yellow Orange Obovoid Obtuse Raised Light brown
Elliptic
TB12 Orange yellow
Orange Obovoid Obtuse Flat Light brown
Elliptic
TB13 Orange Orange Obovoid Obtuse Flat Light brown
Elliptic
TB14 Orange yellow
Orange Obovoid Obtuse Raised Light brown
Elliptic
TB15 Yellowish white
Orange yellow
Obovoid Round Flat Light brown
Elliptic
144
‘Cardona’, ‘Alagrie’ (Llacer et al., 2003), ‘Zhaozhong’ and ‘Baili’ (Feng et al.,
2007) have oblong fruit shape. Magdal and ‘Cardona’ cultivars in Spain have been
observed to have elliptical seed shape, while round seed shape has been noted in
‘Golden Nugget’ and ‘Crisanto Amadeo’ (Llacer et al., 2003).
4.1.8.3 Fruit characteristics
Fruit characteristics of the genotypes are given in Table 47 which shows
the significant difference among the genotypes. Fruit length was maximum (5.08
cm) in TB15 followed with non significant difference by TB5 (5.04). It was lowest
in TB6 (2.88 cm). Fruit width was highest (4.15 cm) in TB15 which was followed
with a significant difference by TB8 and TB13 (both having 3.72 cm width).
Lowest fruit width was observed in TB2 (2.49 cm). Width length index was
highest in TB3 (0.93) followed by TB6 (0.88) and lowest in TB5 (0.71). Fruit
weight was maximum in TB15 (47.84 g) followed by TB8 (46.05 g) while lowest
in TB2 (11.04 g). Fruit volume was maximum in TB15 (45.79 mm3) followed by
TB8 (44.33 mm3). TB2 remained at the bottom with a fruit volume of 10.53 mm3.
Highest flesh seed ratio by weight (3.05) as well as by volume (3.57) was observed
in TB8. It was followed by TB11 which had these ratios as 2.90 and 3.33
respectively. Lowest flesh seed ratio by weight (1.96) as well as by volume (2.31)
was recorded in TB2. Number of fruits per bunch was highest (13.50) in TB1
followed by TB9 (11.23) and minimum in TB14 (6.38). TB3 took the maximum
time (136.2 days) from full bloom to maturity. TB2 and TB 8 remained at par with
it taking 135.0 days and 134.8 days respectively from full bloom to maturity. This
period was lowest in TB13 (117.5 days).
145
Highest yield per tree was recorded in TB7 (89.85 kg) followed by TB5
(69.47 kg) with a significant difference. TB15 had a yield of 25.85 kg per tree and
remained at bottom showing significant difference.
Three genotypes TB3, TB5 and TB7 are the best with reference to yield per
plant which is 57.30 kg, 69.47 kg and 89.85 kg respectively. Yield of the first two
genotypes is comparable with that of ‘Champagne de Grasse’ (70 kg per plant) and
‘M. Marie’ (69 kg per plant) in Turkey (Karadeniz, 2003), while TB7 has a much
higher yield as compared with that of ‘Algerie’ and ‘Gold Nugget’ (Hermoso and
Farre, 2003) which gave a yield of 74 kg and 72 kg per plant respectively in Spain.
Out of the 15 genotypes at Takht Bhai, 8 have been observed to have fruit
weight more than 25g, and three among them (TB8, TB12 and TB15) have fruit
weight even more than 35 g. The highest fruit weight observed in TB15 (47.84 g)
and TB8 (46.05 g) is comparatively greater than that of ‘Magdal’ (45.50 g) and
‘Cardona’ (45.40 g) recorded in Spain (Llacer et al. 2003). It is almost double the
size of ‘Wuerbaisha’, a loquat cultivar in China, which had fruit weight of 24.80 g
(Feng et al. 2007). On the other hand, a number of cultivars in China and Spain
have even higher fruit weight. ‘Zhaozhong 6’ (Zheng, 2001), ‘Mojia No. 1’ (He et
al., 2007), ‘Donghuzao’ (Zhao et al., 2001) and ‘Hongdenglong’ (Jiang et al.,
2001) have fruit weight of 52.1 g, 53.2 g, 59.2 g and 63.1 g respectively. ‘Tanaka’
and ‘Algerie’, cultivars in Spain have fruit weight of 60.60 g and 65.00 g
respectively, while fruit weight of ‘Peluches’ in Spain has been reported as 95.00
g, almost double the size of TB15 and TB8. Anyhow, no cultivar has yet been
observed in Pakistan to have the fruits larger than those of TB15 and TB8.
146
Table 47 Fruit characteristics of 15 loquat genotypes at Takht Bhai
Fruit
characters
Year TB1 TB2 TB3 TB4 TB5 TB6 TB7 TB8 TB9 TB10 TB11 TB12 TB13 TB14 TB15 CV
%
Fruit length
(cm)
Year
I
4.14
c
3.02
fg
3.09
f
3.66 e 5.02
a
2.89
g
3.90
d
4.42
b
3.84
d
4.20 c 4.22 c 3.93
d
4.44
b
3.87
d
5.06 a 2.49
Year
II
4.15
c
3.00
f
3.11
f
3.67 e 5.06
a
2.87
g
3.90
d
4.41
b
3.84
d
4.21 c 4.23 c 3.93
d
4.43
b
3.86
d
5.09 a 1.83
Mean 4.15
C
3.01
F
3.10
F
3.67 E 5.04
A
2.88
G
3.90
D
4.42
B
3.84
D
4.20
C
4.23
C
3.93
D
4.43
B
3.87
D
5.08
A
2.18
Fruit width
(cm)
Year
I
3.16
e
2.49
h
2.88
fg
2.75 g 3.56
c
2.54
h
3.32
d
3.72
b
2.94
f
3.19
de
3.32
d
3.28
de
3.74
b
2.77
g
4.14 a 2.57
Year
II
3.17
f
2.48
i
2.89
g
2.78 h 3.57
c
2.53
i
3.33
d
3.72
b
2.92
g
3.21
ef
3.33
d
3.29
de
3.71
b
2.76
h
4.16 a 1.78
Mean 3.17
E
2.49
H
2.88
F
2.76 G 3.57
C
2.53
H
3.33
D
3.72
B
2.93
F
3.20
E
3.33
D
3.29
D
3.72
B
2.76
G
4.15
A
2.21
WLI Year
I
0.77
g
0.83
de
0.93
a
0.75 g 0.71
h
0.88
b
0.85
c
0.84
cd
0.77
g
0.76
g
0.79 f 0.84
cd
0.84
cd
0.72
h
0.82 e 1.20
Year
II
0.76
h
0.83
ef
0.93
a
0.76 h 0.70
i
0.88
b
0.85
c
0.84
d
0.76
h
0.76
h
0.79
g
0.83
de
0.83
de
0.71 i 0.82 f 0.95
Mean 0.77
H
0.83
EF
0.93
A
0.75 H 0.71
I
0.88
B
0.85
C
0.84
D
0.76
H
0.76
H
0.79
G
0.84
DE
0.84
D
0.72 I 0.82
F
1.08
147
Table 47 Fruit characteristics of 15 loquat genotypes at Takht Bhai (continued)
Fruit
characteristics
Year TB1 TB2 TB3 TB4 TB5 TB6 TB7 TB8 TB9 TB10 TB11 TB12 TB13 TB14 TB15 CV
%
Fruit weight
(g)
Year
I
28.94
d
11.09
h
13.57
gh
16.52
g
22.03
f
13.94
g
26.28
de
46.14
a
14.56
g
25.91
e
26.06
e
36.46
b
33.37
c
16.01
g
47.75
a
6.39
Year
II
29.23
d
10.98
i
13.80
h
16.61g 22.28
f
13.78
h
26.40
e
45.97
a
14.45
gh
26.12
e
26.24
e
36.58
b
33.18
c
15.82
gh
47.93
a
5.39
Mean 29.09
E
11.04
K
13.68
J
16.57
H
22.16
G
13.86
J
26.34
F
46.05
B
14.50
IJ
26.02
F
26.15
F
36.52
C
33.28
D
15.91
HI
47.84
A
5.91
Fruit volume
(mm3)
Year
I
27.43
d
10.55
i
12.89
hi
15.84
g
20.91
f
13.15
ghi
24.86
de
44.44
a
13.96
gh
24.34
e
24.95
de
35.14
b
31.83
c
15.32
gh
45.70
a
6.29
Year
II
27.68
d
10.52
i
13.12
h
15.92
g
21.15
f
13.00
h
24.97
e
44.23
a
13.84
gh
24.56
e
25.13
e
35.27
b
31.62
c
15.12
gh
45.89
a
5.28
Mean 27.56
E
10.53
M
13.01
L
15.88
I
21.03
H
13.08
L
24.92
F
44.33
B
13.90
K
24.45
G
25.04
F
35.21
C
31.72
D
15.22
J
45.79
A
5.81
F:S wt. Year
I
2.81
b
1.98
g
2.32
de
2.15
efg
2.79
b
2..02
g
2.84
b
3.05
a
2.50
c
2.74
b
2.90
ab
2.44
cd
2.21
ef
2.13
fg
2.87
b
3.90
Year
II
2.80
b
1.94
h
2.30
de
2.12
fg
2.77
b
2.02
gh
2.83
b
3.04
a
2.48
c
2.75
b
2.91
ab
2.44
cd
2.21
ef
2.14
efg
2.86
b
3.83
Mean 2.81
BC
1.96
H
2.31
E
2.13
FG
2.78
BC
2.02
GH
2.83
BC
3.05
A
2.49
D
2.74
C
2.90
B
2.44
D
2.21
EF
2.14
FG
2.87
B
3.87
148
Table 47 Fruit characteristics of 15 loquat genotypes at Takht Bhai (continued)
Fruit
characteristics
Year TB1 TB2 TB3 TB4 TB5 TB6 TB7 TB8 TB9 TB10 TB11 TB12 TB13 TB14 TB15 CV
%
F:S vol. Year
I
3.23
bc
2.31
g
2.73
de
2.55
ef
3.19
bc
2.34
fg
3.26
bc
3.57
a
2.89
d
3.10
c
3.33
b
2.82
d
2.54
ef
2.44
fg
3.28
bc
4.07
Year
II
3.19
bc
2.31
f
2.71
cd
2.53
de
3.17
b
2.34
ef
3.24
b
3.56
a
2.88
c
3.11
b
3.33
b
2.83
c
2.53
de
2.45
ef
3.27
b
4.08
Mean 3.21
BC
2.31
G
2.72
E
2.54
F
3.18
BC
2.34
G
3.25
BC
3.57
A
2.89
D
3.11
C
3.33
B
2.82
DE
2.53
F
2.45
FG
3.27
B
4.07
Fruits per
bunch
Year
I
13.73
a
7.63
gh
9.53
de
8.10
fgh
7.17
hi
8.67
ef
10.27
cd
8.53
fg
11.27
b
10.37
bcd
8.83
ef
8.13
fgh
11.20
bc
6.57 i 7.23
hi
5.94
Year
II
13.27
a
7.27
fgh
9.40
cd
7.87
efg
7.13
gh
8.47
e
10.10
c
8.33
e
11.20
b
10.10
c
8.67
de
7.97
ef
10.87
b
6.20 i 6.90
hi
5.12
Mean 13.50
A
7.45
GH
9.47
D
7.98
FG
7.15
H
8.57
EF
10.18
C
8.43
EF
11.23
B
10.23
C
8.75
E
8.05
FG
11.03
B
6.38 I 7.07
H
5.56
Days FB to M Year
I
132.3
c
135.0
abc
136.7
a
124.0
de
126.3
d
124.0
de
119.7
fg
135.7
ab
134.3
abc
133.7
bc
122.0
ef
124.7
de
117.7
g
118.7
g
132.3
c
1.23
Year
II
131.7
c
135.0
ab
135.7
a
123.0
d
125.3
d
123.3
d
118.7
e
134.0
abc
132.3
bc
131.3
c
119.7
e
122.7
d
117.3
e
117.7
e
131.0
c
1.39
Mean 132.0
C
135.0
AB
136.2
A
123.5
E
125.8
D
123.7
E
119.2
FG
134.8
AB
133.3
BC
132.5
C
120.8
F
123.7
E
117.5
G
118.2
G
131.7
C
1.31
149
Table 47 Fruit characteristics of 15 loquat genotypes at Takht Bhai (continued)
Fruit
characteristics
Year TB1 TB2 TB3 TB4 TB5 TB6 TB7 TB8 TB9 TB10 TB11 TB12 TB13 TB14 TB15 CV
%
Yield per tree
(kg)
Year
I
48.15
de
38.62
hi
56.83
c
43.85
efg
68.25
b
42.92
fgh
88.75
a
30.82
jk
51.58
d
40.42
gh
30.32
k
33.82
jk
45.90
ef
35.28
ij
24.65
l
5.82
Year
II
50.58
de
40.37
hi
57.77
c
45.92
fg
70.70
b
45.20
fg
90.95
a
33.33
jk
53.23
d
43.70
gh
32.85
k
36.32
jk
48.60
ef
36.98
ij
27.05
l
4.75
Mean 49.37
E
39.49
H
57.30
C
44.88
FG
69.47
B
44.06
G
89.85
A
32.08
J
52.41
D
42.06
GH
31.58
J
35.07
I
47.25
EF
36.13
I
25.85
K
5.29
WLI=Width length index; F:S wt. = Flesh seed ratio by weight; F:S vol. = Flesh seed ratio by volume; Days FB to M = Days from full bloom to maturity Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
150
TB8, TB11, TB15 and TB7 have excellent flesh seed ratios (3.05, 2.90,
2.87 and 2.83 respectively. ‘Dr. Trabut’, ‘Gold Nugget’, ‘Baffico’ and ‘Kanro’ in
Turkey have been found to have a flesh seed ratio of 3.79, 3.83, 4.16 and 5.42
respectively (Durgac et al., 2006).In Italy, flesh seed ratio in ‘Peluches’,
‘Vainiglia’, ‘Ferdinando’, ‘Tanaka’ and ‘Magdal’ have been found to be 5.9, 5.4,
5.3, 6.4 and 6.5 respectively (Insero et al., 2003).
4.1.8.4 Seed characteristics
Significant differences were noted among the genotypes with respect to all
the seed characteristics studied (Table 48). Number of seeds per fruit was
maximum (5.13) in TB15 which was followed by TB8 (4.88) with a significant
difference. TB6 had the lowest number of seeds per fruit (3.32). Seed weight was
maximum (2.41 g) in TB15 followed by TB12 (2.40 g) both being at par with each
other. TB3 had the lowest seed weight (0.98 g). Seed content per fruit was
maximum in TB15 (12.37 g) followed by TB8 (11.37 g) while minimum in TB2
(3.73 g). It is clear that the genotypes which are at the top with reference to fruit
weight have also the highest number of seeds per fruit as well as seed content per
fruit. In Spain, fruit weight of ‘Italiano 1’ (51.40 g) is not much different from that
of TB15 and TB8 while it has only 3.70 seeds per fruit. Moreover, its seed content
per fruit is just 6.50 g which is almost half the seed content of TB15 and TB8.
Lowest number of seeds per fruit found in TB6 (3.32) is still greater than that of
‘Niuteibaisha’ cultivar in China having 2.78 seeds per fruit (Feng et al., 2007).
Some cultivars with 2 or less seeds per fruit have also been reported. ‘Taicheng 4’
151
(Xie et al., 2007) and ‘White loquat’ (Huang et al., 2007) were observed to have
only 1.32 and 2 seeds per fruit respectively
4.1.8.5 Leaf characteristic
All the genotypes exhibited the significant differences with reference to the
leaf characteristics studied (Table 49). Leaf length was maximum in TB8 (30.16
cm) followed by TB15 (29.98 cm) with a non significant difference, while it was
minimum in TB9 (18.28 cm). Maximum leaf width was recorded in TB15 (9.81
cm). TB5 and TB8 remained at par with it having leaf width of 9.57 cm and 9.75
cm respectively. Lowest leaf width was observed in TB10 (5.89 cm). TB15
remained at top with reference to leaf area (215.43 cm2). TB8 had a leaf area of
214.97 cm2 and remained at par with TB15. TB10 had the lowest leaf area (75.56
cm2). Cultivars with large leaf size are comparable with the Chinese cultivar,
‘Guangyu’ which has the leaves with 28.70 cm length and 10.00 cm width (Feng et
al., 2007). The cultivars with small leaves are comparable with ‘Mojia No. 1’
having 19.80 cm leaf length and 5.20 cm leaf width (He et al., 2007).
4.1.8.6 Inflorescence
All the genotypes significantly differed in floral characteristics (Table 50).
Number of flowers per panicle was highest in TB4 (172.82) followed by TB7
(164.07) while lowest in TB12 (75.15). In literature, different loquat cultivars have
been reported to have different number of flowers per cluster, such as 72 in
‘Zhaozhong’, 92 in ‘Qingzhong’ (Feng et al., 2003), 130.40 in ‘Ferdinando’,
176.20 in ‘Vainiglia’ (Insero et al., 2003), 189 in ‘Golden Nugget’, 227 in
152
‘Buenet’ and 273 in ‘Saval 2’ (Llacer et al., 2003). Maximum panicle size (23.20
cm) was observed in TB1 which was followed by TB13 (22.80 cm) with non
significant difference. TB10 remained at par with TB13 with a panicle size of
22.51 cm. TB6 had the lowest size of panicle (17.99 cm). TB5 took the maximum
time from flowering to full bloom (50.50 days). TB2 followed it with non
significant difference by taking 49.17 days from flowering to full bloom. TB6,
TB12 and TB15 also remained at par with TB5. The period from flowering to full
bloom was shortest in TB9 (39.00 days).
4.1.9 Correlations among some physical traits of loquat genotypes
Study of forty two loquat genotypes showed positive correlations in many
of the parameters analyzed (Table 51). Fruit weight had positive correlation with
seed content (0.983). There is a gradual increase in the fruit weight with the
increase in seed content and number of seeds. Development of embryo in the seed
might be a source of hormones like GA3 that plays a promoting role in
development of fruit.
Fruit weight and seed content had also positive correlation with the flesh
seed ratio (0.657 and 0.525 respectively). Positive correlation of fruit weight with
the flesh seed ratio indicates that larger fruits have more edible portion.
Preliminary findings of this study have already indicated the positive correlation of
flesh seed ratio with fruit weight as well as with seed weight (Hussain et al., 2007).
153
Table 48 Seed characteristics of 15 loquat genotypes at Takht Bhai
Genotypes Number of seeds per fruit Seed weight (g) Seed content per fruit (g)
Year I Year II Mean Year I Year II Mean Year I Year II Mean
TB1 4.03 f 4.10 e 4.07 H 1.88 c 1.88 d 1.88 D 7.59 d 7.69 d 7.64 D
TB2 3.38 i 3.38 h 3.38 K 1.10 gh 1.10 h 1.10 H 3.73 j 3.73 j 3.73 J
TB3 4.22 e 4.22 e 4.22 G 0.97 i 1.00 i 0.98 I 4.09 ij 4.19 hi 4.14 I
TB4 3.85 g 3.85 f 3.85 I 1.36 f 1.38 g 1.37 G 5.25 g 5.32 g 5.29 G
TB5 3.38 i 3.40 h 3.39 K 1.72 d 1.74 e 1.73 E 5.81 f 5.90 f 5.85 F
TB6 3.32 i 3.32 h 3.32 K 1.39 f 1.38 g 1.38 G 4.60 hi 4.56 h 4.58 H
TB7 3.63 h 3.63 g 3.63 J 1.88 c 1.90 d 1.89 D 6.84 e 6.89 e 6.87 E
TB8 4.87 b 4.88 b 4.88 B 2.34 a 2.33 b 2.33 B 11.37 b 11.36 b 11.37 B
TB9 4.10 f 4.15 e 4.13 H 1.01 hi 1.00 i 1.01 I 4.16 ij 4.14 i 4.15 I
TB10 4.32 e 4.35 d 4.33 F 1.60 e 1.60 f 1.60 F 6.92 e 6.97 e 6.95 E
TB11 4.78 bc 4.75 c 4.77 C 1.40 f 1.41 g 1.41 G 6.68 e 6.72 e 6.70 E
TB12 4.43 d 4.42 d 4.43 E 2.39 a 2.41 a 2.40 A 10.60 c 10.62 c 10.61 C
TB13 4.68 c 4.65 c 4.67 D 2.22 b 2.22 c 2.22 C 10.38 c 10.34 c 10.36 C
TB14 4.43 d 4.45 d 4.44 E 1.15 g 1.13 h 1.14 H 5.10 gh 5.03 g 5.06 G
TB15 5.13 a 5.12 a 5.13 A 2.40 a 2.42 a 2.41 A 12.34 a 12.40 a 12.37 A
CV % 1.54 1.86 1.70 3.34 2.50 2.95 4.28 3.15 3.76
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
154
Table 49 Leaf characteristics of 15 loquat genotypes at Takht Bhai
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
Genotypes Leaf length (cm) Leaf width (cm) Leaf area (cm2) Year I Year II Mean Year I Year II Mean Year I Year II Mean
TB1 21.92 d 22.11 d 22.02 D 8.74 cd 8.81 b 8.78 B 138.77 d 140.84 d 139.80 D
TB2 19.94 e 20.21 ef 20.08 EF 6.94 f 7.03 d 6.98 D 89.53 g 91.34 gh 90.43 G
TB3 22.18 d 22.77 d 22.48 D 8.39 de 8.30 c 8.35 C 123.30 e 125.40 e 124.35 E
TB4 18.75 f 18.84 fg 18.80 GH 6.15 g 6.06 ef 6.11 EF 83.58 gh 80.90 hi 82.24 H
TB5 27.87 b 28.45 b 28.16 B 9.53 ab 9.61 a 9.57 A 165.00 b 167.31 b 166.14 B
TB6 21.43 d 22.16 d 21.80 D 8.04 e 8.13 c 8.09 C 126.40 e 129.21 e 127.79 E
TB7 20.29 e 20.65 e 20.47 E 6.32 g 6.42 e 6.37 E 89.51 g 92.28 g 90.89 G
TB8 29.82 a 30.50 a 30.16 A 9.72 a 9.77 a 9.75 A 213.33 a 216.60 a 214.97 A
TB9 18.33 f 18.23 g 18.28 H 6.17 g 6.26 ef 6.22 EF 80.98 gh 82.35 ghi 81.66 H
TB10 19.13 ef 19.57 ef 19.35 FG 5.86 g 5.91 f 5.89 F 74.10 h 77.01 i 75.56 H
TB11 21.49 d 22.04 d 21.77 D 8.86 cd 8.93 b 8.90 B 137.10 d 139.23 d 138.17 D
TB12 24.36 c 24.89 c 24.63 C 7.19 f 7.25 d 7.22 D 110.90 f 113.61 f 112.26 F
TB13 22.43 d 22.48 d 22.46 D 9.15 bc 9.09 b 9.12 B 153.30 c 151.74 c 152.52 C
TB14 22.23 d 22.63 d 22.43 D 7.14 f 7.21 d 7.18 D 107.91 f 110.78 f 109.34 F
TB15 29.72 a 30.23 a 29.98 A 9.77 a 9.85 a 9.81 A 213.75 a 217.11 a 215.43 A
CV % 2.97 3.31 3.15 3.81 3.34 3.58 4.30 4.62 4.46
155
Table 50 Floral characteristics of 15 loquat genotypes at Takht Bhai
Genotypes Number of flowers per panicle Panicle size (cm) Days from flowering to full bloom
Year I Year II Mean Year I Year II Mean Year I Year II Mean
TB1 102.42 hi 109.17 e 105.79 F 23.09 a 23.32 a 23.20 A 42.00 d 44.00 d 43.00 F
TB2 109.22 gh 113.57 e 111.39 F 18.48 ef 18.61 g 18.55 H 48.33 ab 50.00 ab 49.17 AB
TB3 156.83 bc 160.07 b 158.45 B 19.74 c 19.87 e 19.81 EF 38.67 ef 40.67 ef 39.67 G
TB4 169.00 a 176.63 a 172.82 A 19.83 c 19.92 e 19.87 E 41.67 d 42.67 de 42.17 F
TB5 145.80 cd 142.55 c 144.18 C 19.24 cd 19.38 ef 19.31 G 50.00 a 51.00 a 50.50 A
TB6 93.47 ij 96.83 f 95.15 G 17.78 f 18.19 g 17.99 I 48.00 abc 49.33 abc 48.67 ABCD
TB7 164.32 ab 163.82 b 164.07 B 18.44 ef 18.56 g 18.50 H 47.67 abc 48.67 abc 48.17 BCD
TB8 88.93 j 92.02 f 90.47 G 18.72 de 18.77 fg 18.75 H 46.67 bc 47.67 bc 47.17 CDE
TB9 120.10 fg 125.55 d 122.83 E 21.20 b 21.36 c 21.28 C 38.33 f 39.67 f 39.00 G
TB10 103.35 hi 108.47 e 105.91 F 22.42 a 22.61 b 22.51 B 41.00 de 42.00 def 41.50 F
TB11 134.55 de 138.58 c 136.57 D 19.30 cd 19.47 e 19.38 FG 45.33 c 46.67 c 46.00 E
TB12 71.72 k 78.58 g 75.15 H 20.58 b 20.67 d 20.63 D 48.00 abc 49.67 ab 48.83 ABC
TB13 130.58 ef 137.40 c 134.00 D 22.65 a 22.96 ab 22.80 AB 46.00 bc 47.67 bc 46.83 DE
TB14 142.32 d 145.87 c 144.09 C 18.50 ef 18.63 g 18.56 H 41.33 d 42.67 de 42.00 F
TB15 87.08 j 92.08 f 89.58 G 19.48 c 19.72 e 19.60 EFG 48.33 ab 49.33 abc 48.83 ABC
CV % 5.47 4.42 5.00 2.05 1.81 1.93 3.25 3.26 3.26
Means not sharing a letter differ significantly at p < 0.05
Small letters relate to the means of Year I or Year II while capital letters to the combined analysis
156
Table 51 Correlations among some physical traits of 42 loquat genotypes
Fruit weight Seeds / fruit Seed content / fruit
Seed weight Flesh seed ratio
Fruits / bunch
Yield / plant Leaf area Flowers / panicle
Seeds / fruit 0.661
0.000
Seed content / fruit
0.983 0.000
0.661 0.000
Seed weight 0.835
0.000 0.191 0.083
0.853 0.000
Flesh seed ratio
0.657 0.000
0.385 0.000
0.525 0.000
0.467 0.000
Fruits / bunch
-0.326 0.002
-0.424 0.000
-0.320 0.003
-0.151 0.171
-0.228 0.037
Yield / plant 0.021
0.849 -0.076 0.493
0.042 0.702
0.100 0.365
-0.018 0.874
0.626 0.000
Leaf area 0.002
0.986 -0.072 0.514
0.004 0.973
0.011 0.923
-0.048 0.665
0.634 0.000
0.644 0.000
Flowers / panicle
-0.308 0.004
-0.362 0.001
-0.298 0.006
-0.162 0.142
-0.230 0.035
0.902 0.000
0.769 0.000
0.716 0.000
Panicle size -0.171
0.120 -0.234 0.032
-0.159 0.148
-0.060 0.589
-0.132 0.232
0.809 0.000
0.804 0.000
0.747 0.000
0.876 0.000
P-Value < 0.05 shows a significant correlation
157
Flowers per panicle and fruit per bunch had negative correlation (-0.308
and -0.326 respectively) with fruit weight. This is due to the fact that more number
of flowers results in more fruits to set that compete for photosynthates, while the
available food reserves of the plant required for the fruit development are the same.
Hence, the resulting fruits are smaller in size and weight, so having low flesh seed
ratio.
Loquat has a strong tendency to set more fruit. Heavy fruit setting leads to a
high proportion of small sized fruits while fruit size is essential for profitability
(Cuevas et al., 2004). It often makes fruit thinning mandatory to get good quality
fruits. Thinning of flowers and fruits is very helpful practice to get larger fruits in
loquat. Chemical thinning of flowers in loquat reduces the number of fruits per
panicle by 30 percent besides increasing the fruit diameter by 10 percent at harvest
(Agusti et al., 2000). It increases the size and weight of loquat fruit and improves
the fruit quality as well as the edible portion in loquat (Wu and Lin, 2003).
Fruit thinning is a cultural practice to reduce the number of fruits to a level
where a better fruit size can be achieved. It reduces competition among the
developing fruits, consequently increasing fruit ingredients concentration and
appreciably modifying fruit development (Agusti et al., 2003).
Although number of flowers per panicle is negatively correlated with fruit
weight, it has positive correlation with the fruit yield per plant (0.769) because
more flowers result in more fruit set and hence more yield is obtained. Flower
thinning causes a modest reduction in the fruit yield per plant (Agusti et al., 2000)
158
while it is helpful to increase the size, weight and quality of loquat fruit (Wu and
Lin, 2003; Cuevas et al., 2004).
Size of panicle was positively correlated with the leaf area, number of
flowers per panicle and fruits per bunch. It had also positive correlation with the
yield per plant (0.804). These results support the findings of Badenes et al. (2000).
Leaf area had positive correlation with number of flowers per panicle
(0.716), number of fruits per bunch (0.634) as well as fruit yield per plant (0.644).
Increase in leaf area facilitates more photosynthetic activity and hence more
accumulation of food reserves. In the autumn, when temperature is low, vegetative
growth is also slow, while photosynthesis is still going on. This is the flowering
time of loquat while quite sufficient amount of plant food reserves are available for
flowers to bloom and to get fertilized. In this way larger leaves are helpful in better
fruit setting and development that ultimately increases the yield per plant. The
results reveal that leaf area can be one of the potential candidates amongst other
characteristics for selection of better loquat genotypes.
4.2 CHARACTERIZATION OF LOQUAT GENOTYPES ON THE BASIS
OF MOLECULAR MARKERS
Though the selection systems of breeding material on the basis of
morphological characters remain valuable, but this assessment has limitations,
including the influence of environment or management practices (Gepts, 1993;
Nemera et al., 2006). Moreover, the conventional approach to characterize the
cultivars in fruit tree species on the basis of phenotypic observations is slow due to
the long life cycle of plants. Therefore, there is a need to incorporate the new
159
methods based on studies at the DNA level in order to determine the genetic
relationships among different cultivars (Wunsch and Hormaza, 2002; Shiran et al.,
2007).
Problems related with taxonomical classification highlight the need of
complementary keys for identification and characterization of the genotypes. A
standard set of RAPD primers can be established to characterize most of the
common genotypes that may serve as a useful supplement to the traditional
morphological information (Nandini and Chikkadevaiah, 2005). Molecular
techniques cannot replace the characterization for morphological traits, though the
results of molecular studies should be considered as complementary to the
morphological characterization (Karp et al., 1997).
4.2.1 RAPD Analysis
Keeping in view the above facts, the genotypes identified through the
morphological and physical characters were also subjected to the DNA analysis to
determine the level of genetic diversity among the local loquat genotypes found in
different areas of Pakistan and to assess the relationships among them. For this
purpose, DNA extracted from 42 loquat genotypes was amplified using 14 random
primers and run on the agarose gel. Out of 14 decamer RAPD primers used, five
primers generated strong amplifications and resulted in polymorphic products
(Table 52). The remaining nine primers were not considered for compiling the
results because they were either not polymorphic or did not give amplifications.
160
Table 52 Polymorphism revealed by different RAPD primers
Primer Sequence (5- 3) Scored bands
Polymorphic
bands
Polymorphism
rate (%) GL DecamerA-02
TGCCGAGCTG 07 07 100
GL DecamerC-02
GTGAGGCGTC 10 10 100
GL DecamerC-05
GATGACCGCC 12 12 100
GL DecamerC-07
GTCCCGACGA 09 09 100
GL DecamerC-19
GTTGCCAGCC 10 09 90
Total 48 47 97.92 Average 9.6 9.4 97.92 Range 7-12 7-12 90-100
161
The bands obtained through electrophoresis were photographed with the help of
gel documentation system (Fig. 2 to Fig. 6). Out of the 48 amplification products
scored, 47 bands (97.92%) were found to be polymorphic. The average number of
scoreable bands per primer was 9.6, while average number of polymorphic bands
was 9.4 (having a range from 7-12 bands per primer). High frequency of
polymorphism was detected with all the selected primers. The percentage of
polymorphic bands was 100% with 4 primers i.e. GL DecamerA-02, GL
DecamerC-02, GL DecamerC-05 and GL DecamerC-07, while GL
DecamerC-19 exhibited comparatively low level of variability and the percentage
of polymorphic bands was 90%.
Similar results have also been reported by Agar et al., (2008) who used 40
decamer primers to investigate 23 apricot cultivars. However, 28 primers did not
give polymorphic bands or did not amplify clear products. 12 primers produced
good and reproducible polymorphic bands and were used for further analysis. Out
of these 12 primers, 11 produced 100 % polymorphic bands and overall percentage
of polymorphic markers was 97.5.
4.2.2 Cluster Analysis Based on RAPD Markers
Cluster analysis was performed to establish the genetic diversity among the
42 loquat genotypes growing in different areas of Pakistan. The dandrogram was
constructed on the basis of presence and absence of bands by using the software
‘Statistica’ (Fig 7). According to the dandrogram, two main groups ‘A’ and ‘B’ of
the loquat genotypes have been identified having a linkage distance of 33%. All
162
the genotypes growing in Chhattar (CH), Tret (TR), Haripur (HP), Hasan Abdal
and Wah garden (HW) come under the first group, while all the genotypes from
Kalar Kahar (KK) and Choa Saiden Shah (CS) fall under the second group. In
general, all genotypes belonging to a certain location came under any one of the
two groups with the exception of those from Mardan (MN) and Takht Bhai (TB).
Genotypes from Mardan and Takht Bhai were found under both the groups. Out of
the three genotypes of Mardan one, (MN3) falls under the first group and the other
two (MN1 and MN2) under the second group. The maximum number of genotypes
(15) was identified at Takht Bhai. Only two of them (TB2 and TB3) were found to
belong to the first group, whereas all the remaining genotypes to the second group.
One genotype from Takht Bhai (TB11) fell in group B but within the group
appeared as a single solitary line different from all other membersof the group.
Grouping together of the genotypes of Chhattar, Tret, Haripur, Hasan
Abdal and Wah garden is understandable because many families of local people
from these areas are linked to one another with social ties and blood relations, and
have a continuous interaction among them. So exchange of material among these
locations is not surprising. Similarly, genotypes of Kalar Kahar and Choa Saiden
Shah formed a distinct cluster under the group B. It is also due to continuous
exchange of material between the two locations, as both are located in the same
district and linked with trade relations since long.
Grouping of genotypes of Choa Saiden Shah and Kalar Kahar together with
those of Mardan and Takht Bhai was astonishing as they are located very distantly
in two different provinces of the country. But when the socio economic set up of
163
Choa Saiden Shah area was taken into consideration, the situation became clear.
Choa Saiden Shah is an old town of the Punjab province where coal mines are
abundantly found. Majority of the mine workers or labourers belong to the North
Western Frontier Province, to which Mardan and Takht Bhai belong. They have
their settlements in Choa Saiden Shah and its periphery for almost three
generations, while they frequently travel between the two provinces. Moreover,
Khewra, a town adjacent to Choa Saiden Shah, is rich in salt mines and same is the
situation over there. These mine workers might have been a source of transfer of
material at such a long distance.
The maximum number of genotypes (15) as well as the maximum level of
genetic diversity was observed at Takht Bahi. Genotypes from Mardan were
mainly found in group B, however, some members also fell in group A, which
predominantly includes genotypes from Chhattar, Tret, Hari Pur, Hasan Abdal and
Wah. This exchange is, howerve, not unexpected as these areas are socially linked
among one another. Moreover, Mardan as well as the last three of the sites
mentioned above are phydically linked through the old Grand Trunk Road (GT
Road).
While observing the clustering pattern, it is evident that in most of the
cases the genetic diversity level among the genotypes of the areas in close
proximity is very narrow, forming small subclusters with very little linkage
distances. Kalar Kahar and Choa Saiden Shah fall under one cluster. Genotypes
from Haripur come under a small subcluster with very small linkage distance. Low
genetic diversity within certain locations may be due to the reason that there was a
164
very little exchange of loquat material among the distant areas due to poor
infrastructure, lack of proper link roads and poor transport facilities in the past.
Genetic diversity within these locations may be attributed to the cross pollination
and heterozygosity as most of the plants are siblings of the seedling parents.
On the other hand, a wide range of genetic diversity exists at Mardan and
Takht Bhai. It is due to the fact that it is the region of progressive growers of
loquat who have been involved in the commercial cultivation of loquat for the past
many decades. They have made a number of loquat introductions and selections
over the years. Maximum number of genotypes (15) has been observed in a single
orchard of a progressive grower at Takht Bhai.
165
Fig. 2 RAPD pattern of 42 loquat genotypes with Primer GL DecamerA-02: M=Marker; 1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
166
Fig. 3 RAPD pattern of 42 loquat genotypes with Primer GL Decamer C-02 : M=Marker; 1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
167
Fig. 4 RAPD pattern of 42 loquat genotypes with Primer GL Decamer C-05: M=Marker; 1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
168
Fig. 5 RAPD pattern of 42 loquat genotypes with Primer GL DecamerC-07: M=Marker; 1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
169
Fig. 6 RAPD pattern of 42 loquat genotypes with Primer GL Decamer C-19: M=Marker; 1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
170
Fig.7 Clustering pattern of 42 loquat genotypes based on RAPD markers
171
SUMMARY
A number of loquat genotypes are scattered in the loquat growing areas of
Pakistan, while no work regarding the description of these genotypes has ever been
carried out. The present study was conducted to evaluate and characterize
the available genotypes in the main loquat growing areas of Pakistan and to
determine the genetic diversity among these genotypes. Identification of
superior genotypes with better characteristics is a basic step for the
documentation of genetic resources and for establishment of the germplasm
unit. Furthermore, availability of superior genotypes is not possible
without their identification and documentation. Characterization is also
needed to provide a base for planning of future breeding strategies.
For this purpose, 9 sites were selected in the loquat growing areas of
Punjab and NWFP. Forty two genotypes were identified in different locations (5
genotypes at Kalar Kahar (KK), 3 genotypes at Choa Saiden Shah (CS), 5
genotypes at Tret (TR), 3 genotypes at Chhattar (CH), 5 genotypes at Hasan Abdal
and Wah garden (HW), 3 genotypes at Haripur (HP), 3 genotypes at Mardan (MN)
and 15 genotypes at Takht Bhai (TB).
Significant differences were observed with reference to various
characteristics among the different genotypes. Fruit weight of different
genotypes ranged from 9.54 g (in HW4) to 47.84 g (in TB15). Range of
flesh seed ratio was from 1.67 (in HW5) to 3.05 (in TB8). Minimum yield
per tree was 25.85 kg (in TB15), while it was maximum (89.87) in TB7.
172
Differences in other characters including number of seeds per fruit, leaf
length, leaf width, leaf area, size and shape of panicle, fruit skin and flesh
colour, fruit shape etc. were also found to be significant.
Positive correlation was observed between fruit weight and flesh
seed ratio. Fruit weight also had positive correlation with the number of
seeds per fruit as well as the seed content per fruit. Yield was positively
correlated with the leaf area, number of flowers per panicle and number of
fruits per bunch.
RAPD analysis of the genotypes was also performed. Five RAPD
primers gave reproducible results and generated 47 polymorphic bands.
According to the dandrogram, two main groups ‘A’ and ‘B’ of the loquat
genotypes were identified having a linkage distance of 33%. For most of the
locations, grouping of the genotypes was in accordance with the geographical
locations. All the genotypes growing in Chhattar, Tret, Haripur, Hasan Abdal and
Wah garden came under the first group, while all the genotypes from Kalar Kahar
and Choa Saiden Shah fall under the second group. Out of the three genotypes of
Mardan, one falls under the first group and the other two under the second group.
The maximum number of genotypes (15) was identified at Takht Bhai. Two of
them were found to belong to the first group and 13 to the second group. Grouping
of the genotypes from Choa Saiden Shah and Kalar Kahar (Punjab Province) with
those from Mardan and Takht Bhai (NWF Province) may be due to transfer of
material through the mine workers of NWFP who have their settlements at Choa
Saiden Shah and its periphery for almost three generations.
173
Genotypes with good characteristics i.e. better yield (TB7, MN3, TB5,
MN2, MN1, TB3, HP3, TR4), higher fruit size and weight (TB15, TB8 MN2,
TR4, MN3, TB12), less number of seeds per fruit (KK5, CH1, HW1), smaller
seeds (TR5, HW3, TB3, KK3, TB9) and higher flesh seed ratio (TB8, TB11,
TB15, TB7, TB1, TR4, TB5, MN2) can be recommended for further multiplication
and introduction to the other loquat growing areas. Statistics show that the loquat
production of the NWFP is much higher than that of the Punjab province, although
area under loquat in the Punjab is comparatively greater than that of NWFP. It is
all due to the occurrence of better genotypes in NWFP. Multiplication of the better
genotypes through vegetative means with a planned programme can be very useful
for the grower. It may increase the per acre return of the farming community as
well as the overall production of the country.
This study may also provide a good support to the efforts of United Nations
and the Government of Pakistan in poverty alleviation. Cultivation of loquat is
quite labour intensive (Lin et al., 2007) and has the capacity to absorb the jobless
manpower in various processes like nursery management, cultural operations, plant
protection measures, harvesting, transport and marketing of loquat. During the
period from late March to April, loquat gives a good sustain to all the links
involved in the production, marketing and sale of fresh fruits as no other fresh fruit
is available.. It also meets with the consumers’ demand of fresh and nutritious fruit
during the time when the citrus fruits have just disappeared and the other fresh
fruits are yet to come in the market. Hence it fetches good prices in the absence of
other competitors. Moreover, availability of superior genotypes, and increased
174
production would lead to export of this fruit to other countries and increase in the
foreign earning of the country.
The study recommends establishing germplasm units in Punjab and NWFP
and pooling all these genotypes for future strategies and breeding programs
including selection, introduction, hybridization and mutation breeding. Area under
loquat can be further increased by extending its cultivation to the districts located
in central part of Punjab.
Future Research
In future, studies can be conducted on the degree of adaptability, yield and
performance of these genotypes in different locations of the country. Research on
the cultural and management practices as adopted in the other countries like China,
Spain and Turkey can be accomplished in our environment as no such research has
ever been conducted so far in loquat. Presently plant height observed is about 25 to
35 feet or even more, which makes the cultural practices including plant protection
and fruit picking very difficult. High density plantation using dwarf rootstock may
increase the yield per unit area. Protected cultivation of loquat can increase
earliness by up to 20 days (Polat and Caliskan, 2007). Use of seedling rootstock
may result in larger tree with high canopy. Grafting on quince can produce early
bearing dwarf trees (Shih, 2007). In future dwarf plants may be developed through
pruning training and other practices as is done in the other loquat producing
countries. Training of the loquat growers and nurserymen is also an important
aspect which requires our attention. These measures would be helpful in
harnessing the maximum potential of our genetic resources.
175
Conclusions
The study has generated valuable information for the Horticulturists,
breeders, nurserymen and the loquat growers. It would be helpful for the
documentation, management and conservation of loquat genetic resources of
Pakistan. Registration of the loquat genotypes, establishment of loquat germplasm
units and availability of true to type plants can play a significant role in improving
the loquat production in the country that will lead to better returns for the growers.
The results of this work will also provided a good foundation for future research on
this crop in our country.
176
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Appendix-1 List of loquat genotypes included in study S. No. Location Codes No. of genotypes 1 Kalar Kahar KK 5 2 Choa Saiden Shah CS 3 3 Chhattar CH 3 4 Tret TR 5 5 Hasan Abdal HW 2 6 Wah HW 3 7 Haripur HP 3 8 Mardan MN 3 9 Takht Bhai TB 15
209
Appendix-2 Area and production of loquat in different countries Country Area ‘000’ hac Production ‘000’ tonnes Yield per hactare
China 120.00 460.00 3.83 Spain 3.023 43.30 14.32
Turkey 0.82 12.00 14.63
Pakistan 1.501 10.479 6.98 Japan 2.42 10.24 4.23
Italy 0.660 4.410 6.68 Brazil 0.300 2.400 8.00
210
Appendix-3 Area and production of loquat in Pakistan during last 5 years Year Area (hactares) Production (tonnes)
2004 1376 9868
2005 1407 10042
2006 1429 10171
2007 1472 10688
2008 1501 10479
211
Appendix-4 Area and production of loquat in provinces of Pakistan Province Area (hac) Production (tonnes) Yield (tonnes per hactare)
Punjab 764 4,687 6.14
NWFP 657 5,809 8.84
Baluchistan 51 192 3.76
212
Appendix-5 Loquat germplasm resources in different countries Country Site No. of cultivars
China Fruit Research Institute, Fuzhou 250
Spain Institute of Agricultural Research, Valencia 100
Japan Experimental Station of Fruit Trees, Nakasa 60
Italy Palermo University, Sicily 16
213
Appendix -6 Binary matrix of 42 loquat genotypes as obtained by Primer GL DecamerA-02
Band No.
Genotypes1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42
1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 1 0 1 1 0 0 1 1 1 1 1 0 1 1 1 1
2 1 1 1 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 1 0 1 1 0 0 1 1 1 1 0 0 0 0 0 0
3 1 1 1 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 1
4 1 1 1 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 1 0 0 1 0 0 1 1 1 1 0 1 0 0 0 1
5 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0
6 0 0 0 0 0 0 0 0 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 0 0 1 0 0 1 0 0 1 1 1 1 0 1 0 0 0 1
7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
214
Appendix -7 Binary matrix of 42 loquat genotypes as obtained by Primer GL DecamerC-02
Band No.
Genotypes
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42
1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0 1 0 0 0 0 0
2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0
3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1
4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0
5 1 1 1 1 0 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 0 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1
6 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1
7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0
8 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
9 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 1 0 0 1 0 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0
10 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 1 0 0 1 0 1 0 1 0 1 1 0 0 0 0 0 0 0 0 0 1 1 0
1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
215
Appendix -8 Binary matrix of 42 loquat genotypes as obtained by Primer GL DecamerC-05
Band No.
Genotypes
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42
1 0 1 0 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
2 0 1 1 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
3 0 1 1 0 1 1 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0
4 0 1 1 0 1 1 1 0 0 0 0 0 1 1 0 0 1 1 1 0 0 1 1 1 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0
5 1 1 1 1 1 0 0 1 1 1 0 0 1 1 0 0 1 1 1 0 1 1 1 1 0 0 1 0 1 1 0 1 0 0 0 0 0 0 0 0 0 0
6 1 1 1 0 0 1 1 0 1 1 1 0 1 1 1 1 1 1 1 0 1 1 1 1 0 0 1 0 1 1 0 0 0 0 0 0 0 0 0 1 0 1
7 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 1 1 1
8 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 0
9 1 0 1 0 0 1 0 1 1 1 1 0 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 0 1 1 1 1 1 1 0 0 1 1 1 1 1 1
10 0 0 0 1 1 1 0 1 0 1 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 1 1 1 1 1 0 0 0 0 1 1 1 1
11 1 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 1 0 0 1 0 0 0 0 1 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0
12 0 0 0 0 0 0 0 0 0 1 1 0 1 0 1 1 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 0 0 1 0 1 0 0 1 0 0
1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
216
Appendix -9 Binary matrix of 42 loquat genotypes as obtained by Primer GL DecamerC-07
Band No.
Genotypes
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42
1 1 0 0 0 0 0 1 0 1 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1
2 0 0 0 0 0 0 0 0 1 1 0 1 1 1 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
3 1 0 0 1 1 0 1 0 1 0 0 1 1 1 0 0 0 0 0 0 0 1 1 1 1 1 0 0 1 1 1 1 1 1 1 1 1 0 1 1 1 1
4 1 1 1 1 1 1 1 1 1 1 0 1 1 1 0 1 1 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1
5 1 0 1 0 0 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
6 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1 1 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
7 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
8 0 0 0 0 0 0 0 0 1 0 0 1 0 1 0 1 0 0 0 1 1 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
9 0 0 0 0 0 0 0 0 0 0 1 0 1 0 1 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
217
Appendix -10 Binary matrix of 42 loquat genotypes as obtained by Primer GL DecamerC-19
Band No.
Genotypes
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42
1 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 1 1 1 1 0 0 1 1 1 1 1 0 1 0 1 1 1 1 0 1 0 1 0 1 0 1 1
2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 1 1 1 1 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1
3 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 1 1 1 1 0 1 1 1 1
4 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 1 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0
5 1 0 0 0 0 0 0 1 1 1 0 0 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0
6 0 0 1 0 1 1 1 1 1 0 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1
7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 1 1 1 0
8 0 0 0 0 0 1 0 0 1 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 0 1 1 1 1
9 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 1 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0
1=KK1; 2=KK2; 3=KK3; 4=KK4; 5=KK5; 6=CS1; 7=CS2; 8=CS3; 9=TR1; 10=TR2; 11=TR3; 12=TR4; 13=TR5; 14=CH1; 15=CH2; 16=CH3; 17=HP1; 18=HP2; 19=HP3; 20=HW1; 21=HW2; 22=HW3; 23=HW4; 24=HW5; 25=MN1; 26=MN2; 27=MN3; 28=TB1; 29=TB2; 30=TB3; 31=TB4; 32=TB5; 33=TB6; 34=TB7; 35=TB8; 36=TB9; 37=TB10; 38=TB11; 39=TB12; 40=TB13; 41=TB14; 42=TB15.
