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SV:M:MJI_1{cy e{l CO:NCLVSIO:JV
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SUMMARY
In India our divine Vedas were compiled much before the
commencement of the modern era. They embody a vast of knowledge regarding
multifarious uses of plants with major emphasis on medicinal, mcgico-medicinal,
magico-religious and mythological aspects. The Rigveda (3,500-1 ,800 BC)
appears to be the oldest record available on the medicinal plants. Similarly the
Vrikshayurveda compiled by saga Parasara consists of excellent information about
plants and their medicinal properties. (Sensarma, 1989). Ayurveda (about 2500
B.C.) contains a more detailed account of many drugs and their uses. Ayurveda in
fact is the foundation stone of the ancient medicinal science of life and art of
healing. The Charak and Sushrut Samhitas were written between 700-200 BC,
which includes the sum and substance of discovery about medicinal plants by
cowherds, goatherds, shepherds and tribes treasure.
Many of the crude drugs which are sources of medicinal preparations
still come from wild growth. The natural resources how- so - ever large are bound
to diminish. Time has therefore come to bring these plants under plough to meet
the rising demand of the resultant product. These plants, as a group, have a large
potential for primary introduction of new commercial crops. There is an increasing
demand for plant based drugs, and pharmaceuticals in the world market. India is
among the traditional producers and exporters of several medicinal plants. There
is, therefore. need for intensive agricultural studies leading to genetic
improvement and cultivation method for expansion of area under medicinal and
aromatic plants.
Some plants have been cultivated since time immemorial (e.g. Flax,
Opium, Poppy and Coca). Others are now grown because supplies of the wild
plants are insuf!icient to meet the demand or because, owing to sparse distribution
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due to in ability to access, collection is difficult. Hence cultivation is essential in
the case of such drugs as Indian hemp and opium. which are subject to
government control, and recently for those wild plants in danger of over
exploitation and which have now been CITES (q.v.) listing. In many cases,
cultivation is advisable because of the improved quality of the drug, which it is
possible to produce.
There are constrains in commercial cultivation of medicinal plants in our
country. Very little is known about seed biology of medicinal plants and no efforts
have been made to search for and propagate elite specimen. Standard propagation
techniques are available for only a few of the species. There is need for scientific
approach towards propagation germination behavior, branch cutting, growing
mediums, environmental factors etc of medicinal plants. Lack of reliable and
standardized technology package has been bottleneck in popularizing the
commercial cultivation and utilization of important medicinal plants. There is
need to develop agro technology package for each important plant species so that
it could be easily adopted by rural communities.
The growth and development of plants is regulated by a number of chemical
substances which together exert a complex interaction to meet the needs of plant
Five groups of plant hormones are well established: they are the auxins,
gibberellins, cytokinnis, absciscic acid and its derivates, and ethylene. Effect of
these substances vary on the production of secondary metabolites, particularly
with a view to producing plants containing an enhanced proportion of active
constituents. For commercial purposes yield per hectare is an obvious criterion,
whereas for biosynthetic studies yield per plant or percent fresh weight may be of
more significance.
Some of the important experimental works on cultivation of medicinal
plants are: Chatterjee and Lama (1977), described cultivation trials of Cinchona.
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Gadwal ( 1977) has worked on commercial cultivation of Solanum crop. Sen
sarma ( 1989) studied cultivation and utilization of sandal wood. Shukla (1991) has
described twenty-six medicinal plants with their cultivation method. I3isen (1994)
studied the genetic variability and cultivation practices of Safed musly
(Chlorophytum sp.). Kanjilal er a/. (2000) have conducted experiment for the
cultivation of Wedila eslendulaceae at J01·hat, Assam.
Cultivation teclmology has been standardized for menthol mint
(Mentha arvensis) by Kittimani eta!. (2000). Pandey et al. (2003) and Sharma et
a/. (2003) have worked on prospects of Kalmegh (Andrographis paniculata) and
Ashwagandha ( Whithania somnifera) cultivation in Chhattisgarh. Sundriyal (2005)
has studied the medicinal plant cultivation and conservation in the Himalaya. Gera
et al. (2005) have studied the economics of cultivation of some commercially
important medicinal plants.
Pal (1990) has studied the seasonal variation of the effect of auxins
on rooting branch cutting of Hibiscus rosasinensis. Rooting branch cutting of
Solanum hispidum for raising clonal crops were reported by Badola et al. (1991).
Pal eta! (1994, 1995), observed effect of auxin on rooting branch cutting of
Ashwagandha (Whithania somnifera) and vegetative propagation of Sarpagandha
(Rauolfia serpentina). Thakur eta/. (1999) have studied the effect of season and
auxin concentration on rooting and sprouting of Alnus nitida End!. Sundharaiya et
a/. (2000) have recorded the effect of growth regulators in the propagation of
Sarkarikolli (Gymnema sylvesrtre), medicinal Coleus (Coleus forskohlii) and
Thippli (Piper longum). Vandana & Bhatt (2001) studied the propagation of
Sesbania sesban and S. grandiflora through stem cuttings. Jadhav et a!. (2002)
observed the effect of different growth regulators on vegetative propagation of
Patchouli (Pogostemon cab/in Benth). Vegetative propagation of Colutea
nepalellSis Sims, through stem cuttings was studied by Sharma et a/. (2004).
Vegetative propagation of Albizia lebbek through stem cutting was studied by
9!
Handa eta/. (2005). Ramula el a/. (2005) have studied the vegetative propagation
of Hemidermus indicus R.Br. through stem cuttings. IAA 2000 ppm showed best
rooting response followed by IBA and NAA.
A tal & Kapur ( 1982) had recommended a fertilizer mixture of sup a
phosphate, murate of potash and urea each @ 50 kg /ha in medium type of soil for
better growth and yield of Solanum khasainum. Int1uence of Nitrogen on crop
duration seed yield and its components in Glory lily (Gloriosa superba L.) wa>
studied by Kumaraswamy et al. (1994). Masoodi et a/.(1996) studied the effect of
nitrogen (0, 40, 80, 120 kg /ha) and spacing (10 x IDem, 10 ><15cm, 15 xl5cm and
20 x 20cm) on growth and development of Acer oblongum Wall. Seedlings.
Tiwari eta!. (2000-200 1) had determined the effect of planting date on growth and
root productivity dates of Plumbago zeylanica. Mathur et al. (2003) reported
growth and yield of Cumin (Cuminum cyminum L.) as influenced by different
organic manure and biofertilizers. Effect of inter and intra-row plant spacing ( 45 x
30cm, 45 x 45cm, 60 x 30cm, 60 x 45cm and 60 x 60 em) on growth and yield of
wild egg plant or Kateli (Solanum surratense) was studied by Khandelwal et al
(2004 ). Kumar et al. (2005) studied the effect of nitrogen, sulphur and Neem cake
on ,biomass and yield of active constituents, quercetin in a medicinal plant Pluchea
lanceolata.
OBJECTIVES
• Selection of two medicinal herbs Boerhaavia diffusa and Solanum
xanthocarpum.
• To determine suitable soil for the growth of plants.
• To determine the effects of chemical fertilizer and biological manure on
plant production.
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• To determine the effect of hormones on the growth as : Height/length, leaf
size, number of leaves, number of nodes and internodes, number of
branches. number of 11owers and fruits ..
• Estimation of primary metabolites (Carbohydrates and Proteins).
• Estimation of chlorophyll content.
• Estimation of alkaloid present in the berries of Solanum xanthocarpum.
• Estimation of alkaloid present in the roots of Boerhaavia diflusa.
MATERIALS & METHODS
Experiments were conducted to determine the best soil, to examine
the effect of hormones (IAA & IBA) in different concentrations, and to examine
the response of nitrogen, phosphorus and potash fertilization on the maximum
plant growth, chlorophyll content, production of primary metabolites (i.e. ;
carbohydrates & proteins) and alkaloids in the root of Boerhaavia diflusa and
berries of Solanum xanthocarpum.
DETERMINATION OF SUITABILITY OF SOIL:
Three types of the soil Kanhar, Bhata and Matasi were used for the
determination of suitability of soil.
Planting Material:
The seeds of B. diflusa are very small and create difficulties to handle
properly. So the stem cuttings were used as planting material for B. diflusa while
for as S. xanthocarpum seed were used as the planting material.
Pot Preparation:
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30 pots were taken for each of the two species, S. xanthocarpum and B.
dif!usa. 4 kg of soil were used to fill each of the pots I 0 pots with Kanhar soil, I 0
pots with Matasi soil and I 0 pots with Bhata soil.
Planting B. dijfusa:
Shoots of B. dij]i.1sa were collected in the month of July from the campus of
Pt. Ravishankar Shukla University, Raipur. Leaves and apical soft portion of the
plants were excised and then cut in to !5-20cm long pieces. One or two such
cuttings were planted in each pot.
PlantingS. xanthocarpum:
Seeds of S. xanthocarpum were collected during the months of May to June
in every year during the investigation period (2002-2004). Ripe fruits were dried
and seeds were stored in sealed polythene bags at room temperature. Plants were
raised in Botanical garden of School of Life Science, Pt. Ravishankar Shukla
University, Raipur. Collected seeds were sown in nursery beds in month of
December-Janurary. Beds were watered daily to avoid desiccation. Seedlings were
transplanted to potsin the month of Feb-March. Seedling attended a height of
about 4-Scm, after about one month of sowing i.e. in the month of March.
Seedlings of equal size were now taken for planting in the pots, each pot had one
or two seedling per pot.
HORMONE TREATMENT:
Hormonal treatment was given to the cuttings both m B. diffusa & S.
xanthocarpum.
Preparation of Cuttings for B. diffusa and S. xanthocarpum:
Shoots were collected from the plant of B. diffusa in the month of July and
S. xanthocarpum in the month of March .. Their Leaves and apical soft portion of
the plants were excised and cut in to 15-20cm long pieces.
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Treatment and Planting of B. diffusa and S. xantlwcarpum:
The cullings were dipped in 0.1% water suspension of Bavistin for 15
minutes. These cuttings were. then, treated with I 0, 50 and I 00 ppm of indole
acetic acid (II\/\) and indole butyric acid (IBA). Control cuttings were treated with
distilled water by dipping 2 em basal portion of the cuttings for 24 hour's. After
the treatment. cuttings were planted in earthen pots filled with soil and sand in 2: I
ratios. These were then kept under partial shade in a net house and were watered
frequently to avoid desiccation.
FERTILIZER TREATMENT:
Land Preparation:
Experiment on fertilizer treatment was conducted during 2003-2004 with
six treatment combinations as cow dung 10 ton/ha + N:P:K 60:40:20 kg/ha, cow
dung lO ton/ha + N:P:K 45:30:15 kglha, cow dung 10 ton/ha + N:P:K 30:20:10
kglha, and cow dung 5 ton/ha + N:P:K 60:40:20 kglha, cow dung 5 tonlha +
N:P:K 45:30:15 kg/ha and cow dung 5 ton/ha + N:P:K 30:20:10 kg/ha replicated
three times to evaluate the effect of fertilizer combination on growth of plants,
carbohydrates and proteins, and amount of alkaloids in the root of Boerhaavia
diffusa and berries of S. xanthocarpum. The Plot area was 2mx2m for each plot for
both the species. The Soil was pretreated with 10% BHC and Bavistine 0.1% w/w
to avoid insect attack and fungal infection.
Nitrogen was applied in the form of urea (46% N), phosphorus, in the form
the single super phosphate (16% P20s) and basal dose of potash in the form of
murate on potash (60% K20). Control, without N, P and K and compost as cow
dung treatments were also prepared simultoneously.
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Planting B. tfiffusa:
Collected shoots were treated with 0.1% llavistin for 15 minutes. These
treated cuttings were planted in to the experimental plot area at a spacing of 20 x
20cm row to rol\".
Planting S. xanthocarpum:
Collected seeds were sown in Nursery beds to raise the seedlings in the
month of Jan-Feb. Seedling attended a height about 4-Scm, after one month of
sowing i.e. in the month of March. Seedlings of equal size were now taken and
transplanted from nursery to experimental plot area. Twenty five seedlings were
planted in each of the plot area, spacing at 20 x20 em in row to row.
Recording Plant Parameters:
Plant height was measured from ground level to the apex of the main stem,
number of leaves, number of nodes and internodes were recorded for both the
species at monthly intervals and number and length of root were recorded only for
B. diffusa. Number of flower and fruit were recorded only for S. xanthocarpum
be2ause flower and fruits of B. diffusa are very small and hence were not recorded.
Estimation of Primary Metabolites:
Plant extract was prepared by homogenizing 1 gm of plant tissue with 5 ml
of sodium-phosphate buffer (pH-7.2) with mortar and pestle and filtrate obtained
was centrifuged at 15,000 rpm for 20 min at 4°C. Finally, supernatont only was
used for the estimation of total sugars according to Sipro (1966), reducing sugar
according to Wood et al (1988), soluble proteins and TCA precipitated proteins
according to Lowry et a!. ( 1991). Quantitative estimation of chlorophyll was made
following Arnon ( !944) while carotenoides were determined following Duxbury
& Yentsh (1956).
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Extraction of Alkaloid:
Powdered root samples (20mg) of B. dijfusa and powdered green berries of
S. xa11thocarpwn of all the variously treated plants species were extracted with
ethanol in Soxhlet extractor for 24 hours. Alkaloid was confirmed all the different
entire extracted samples by Dragendorffs reagent and Mayer's and Wagners
reagent.
RESULTS
Determination of suitable soil, effect of hormones and different fertilizer
combinations on growth of plant (i.e.; height/length, number of leaves, size of
leaves (length & width), number of nodes and internodes, number of branch,
number of root and length of root and estimation of carbohydrates and proteins,
chlorophyll content and amount of alkaloid in root of B. dijfusa and berries of S.
xanthocarpum were studied. Results of these studies were as follow:
DETERMINATION OF SUITABLE SOIL FOR BOERHAAVIA
DIFFUSA:
Plant grown in Kanhar soil gave best response for almost all the growth
parameters as compared to plant grown in Matasi and Bhata soil. Cuttings grown
in Kanhar soil showed maximum plant height (104.1±10.9) em, number of leaves
(90.66±11.01). size of leaves (length & width) (3.41±0.104 em & 2.61±0.147 em),
number of nodes and internodes (49.6±3.265 & 48.6±3.26), number of branch
(28.87±12.94), number of roots (8.24±0.169) and length of root (26.52±0.256) em
these values were followed by values for cuttings followed by cutting grown in
Matasi and Bhata soil in the same sequence.
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Estimation of total sugar in cutting of B. diffusa grown in Kanhar,
Bhata and Matasi soil :
Maximum amount of total sugar ( 11.53±0.233 mg/g. 9.4±0.25 I mg/g and
8.76±0.176) mg/g was found in root, leaf and stem of cuttings grown in Kanhar
soil. followed by cutting grown in Bhata soil (I 1.53±0.233 mg/g. 8.23±0. I 15 mg/g
and 7.4±0.066 mg/g) in root, leaf and stem respectively and minimum amount of
total sugar was found in roots of cutting grown in Matasi soil with
(I 0.23±0.033mg/g) and in leaf and stem the amount of total sugar was
(7.06±0. 753 mg/g) and (7 .06±0.666 mg/g) was also equal.
Estimation of reducing sugar in cuttings of B. diffusa grown in Kanhar,
Bhata and Matasi soil :
Maximum amount of reducing sugar (3.166±0.033 mg/g, 2.5±0. 13 mg/g)
was found in root and leaf respectively of cuttings grown in Kanhar soil as
compared to (2.86±0.317 mg/g, 2.4±0.0lmg/g and 2.533± 0.066 mglg,
2.03±0.033) mg/g in root and leaf respectively of cutting grown in Matasi and
Bhata soil, but in stem reducing sugar was maximum (1.63±0.88lmg/g) maximum
in cutting grown in Matasi soil as compared to cutting grown in Kanhar and Bhata
soil, respectively.
Estimation of total soluble protein in cuttings of B.diffusa grown in
Kanhar, Bhata and Matasi soil:
Maximum amount of total soluble protein was (13.76±0.033 mg/g,
10.03±0.233 mg/g) and (6.33±0.956 mg/g) in root, leaf and stem respectively in
cutting grown in Kanhar soil followed by cutting grown in Matasi soil with
(10.5±0.404 mg/g, 9.26±0.592 mglg and 5.9±0.152 mg/g) and Bhata soil
(9.9±0.152 mg/g, 8.6±1.02 mg/g and 4.3±0.633 mg/g) in root, leaf and stem
respectively.
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Estimation of TCA precipitated protein in cuttings of B. diffusa grown
in Kanhar, Bhata and Matasi soil:
Maximum amount of TCA precipitated protein (3.2±0. I 73 mg/g and
3.0±0.504 mg/g) was found in root and leaf of cutting grown in Kanhar soil but in
stem it was maximum (3 .I ±0.208 mg/g) in cutting grown in Bhata soil followed
by the amount of TCA precipitated protein which was similar in root and leaf of
cutting grown in Matasi and Bhata soil but was minimum in root (2.4±0.288 mg/g)
in plant grown in Matasi soil.
Estimation of chlorophyll content in cuttings of B. diffusa grown in
Kanhar, Bhata and Matasi soil:
Maximum amount of chlorophyll a, chlorophyll b, total chlorophyll and
carotenoides were (0.416±0.009 mg/g, 0.136±0.006 mg/g, 0.5463±0.008 mg/g and
0.312±0.11mg/g) found respectively were found with cutting grown in Kanhar soil
followed by cutting grown in Matasi soil (0.247±0.008 mg/g, 0.076±0.004 mg/g,
0.3276±0.036 mg/g, 0.2173±0.007 mg/g) and Bhata soil (0.248±0.036 mg/g,
0.0583±0.022 mg/g, 0.304±0.01 mg/g and 9.06±0.014 mg/g) respectively.
Amount of alkaloid in roots of B. diffusa grown in Kanhar, Bhata and
Matasi soil:
Maximum amount of alkaloid was (0.02015 gmlgm) found in root of cuttings of B.
diffusa grown in Kanhar soil followed by cutting grown in Matasi and Bhata soil
with (0.01915 gm/gm) and (0.01814 gm/gm) respectively.
HORJVIONR TREATMENT:
IBA treated cuttings showed better response as compared to IAA
treated cuttings of all the parameters. IBA 50ppm treated cuttings showed
maximum plant height (59 .86± 7.758 em), number of leaves (193. 7± 77.77), size of
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leaves (length & width) (3.346±0.17Icm & 2.799±0.205 em), number of nodes
(52.93±10.98). number of internodes (5 1.93± 10.989), number of branch
( 17 .36±3.302), number of roots (14±2.489) and length of root (56.22±1.612 em )as
compared to IBA !Oppm, IOOppm and IAA lOppm, 50ppm lOOppm treated
cuttings and control.
Estimation of Primary Metabolites:
Estimation of total sugar in cuttings of B. diffusa treated with 10, 50
and IOOppm ofiAA and IBA:
The amount of total sugar was maximum (17.23±0.384 mg/g, 13.8±0.688
mg/g) and (12.0±0.08 mg/g) was found in root, leaf and stem respectively of
SOppm treated cuttings of IBA and second highest amount of total sugar in IAA
50ppm treated cuttings (15.5±0.907 mg/g, 12.76±0.328 mg/g and 11.76±0.08
mg/g) in root leaf and stem respectively IAA !Oppm and IBA!Oppm treated
cutting showed almost equal amount of total sugar in root leaf and stem
respectively as compared to control.
Estimation of reducing sugar in cuttings of B. diffusa treated with 10,
50 and 1 OOppm of IAA and IBA:
Maximum amount of reducing sugar was (3.63±0.08 mg/g, 2.0±0.057
mg/g and 1.76 ±0.133 mg/g) found in root, leaf and stem respectively in IBA
SOppm treated cuttings, followed by IAA 50ppm treated cuttings with (2.63±0.066
mg/g, 1.9±0.11 5mg/g and 1.67±0.15 mg/g) in root leaf and stem respectively.
Control had values (1.33±0.145 mg/g, 1.46±0.98 mg/g and 1.1±0.03 mg/g) in root
leaf and stem respectively.
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Estimation of total soluble protein in cuttings of B. diffusa treated with
10, 50 and IOOppm ofiAA and IBA:
Maximum amount of total soluble protein was (26.9±0.848 mg/g and
21.26±1.36 mg/g) found with root and leaf respectively of IBA 50ppm treated
cutting but in stem with it was maximum (16.33±1.185 mg/g) in IBA!OOppm
treated cuttings. This was followed by IAA I OOppm treated cuttings the amount o
total soluble protein was (23.66±0.480 mg/g) in root and IAA 50ppm treated
cutting showed the amount of total soluble ( 15.96±1.07mg/g and 11.9±0.351mg/g)
in leaf and stem respectively.
Estimation of TCA precipitated protein in cuttings of B. diffusa treated
with 10, 50 and 100ppm ofiAA and IBA:
Maximum amount ofTCA precipitated protein (5.46±0.6 mg/g, 3.92±0.763
mg/g) in root and leaf respectively ofiBA 50ppm treated cutting but in stem it was
maximum (4.8±0.33mg/g) in IBA 100ppm treated cuttings followed by IAA
100ppm treated cutting with the amount of total soluble protein (4.5±0.775 mg/g,
3.53±0.001 mg/g and 2.56±0.417mg/g) in root, leaf and stem respectively. Control
had values (3.26±0.504mg/g, 2.8±0,202 mg/g and 2.8±0.173) mg/g in root, leaf
and stem respectively.
Estimation of chlorophyll content in cuttings of B.diffusa treated with
10, 50 and 100ppm ofiAA and IBA:
Maximum amount of chlorophyll a, chlorophyll b, total chlorophyll and
carotenoides in sequence of (0.604±0.015 mg/g, 0.240±0.031 mg/g, 0.844±0.03
mg/g, 0.513±0.312 mg/g) was found with IBA 50ppm treated cutting followed by
IAA lOOppm treated cutting with (0.60!±0.0381mg/g, 0.135±0.068 mg/g,
0.737±0.428 mg/g, 0.432±0.250 mg/g) as compared to (0.305±0.010 mg/g,
0.111±0.008 mg/g, 0.416±0.019 mg/g and 0.303± 0.012 mg/g) of Chlorophyll a,
Chlorophyll b, total chlorophyll and carotenoides in control.
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Amount of alkaloid in roots of B. diffusa treated with 10, 50 and 100
ppm of IAA and IBA:
Maximum amount of alkaloid (0.07123 gm/gm) was found with IBA
50ppm treated cutting followed by IAA 50ppm treated cutting (0.04981 gm/gm)
and minimum amount of alkaloid was 0.3211 gm/gm in IBA I Oppm treated
cutting as compared to (0.262lgm/gm) in control.
FERTILIZER TREATMENT:
Maximum plant length (123.10±37.7 em), number of leaves
(321.73±82.70), size of leaves (length & width) (3.83±0.208 em & 3.66±0.18cm)
number of nodes and internodes (120.23±44.80 & 119.23±49.80), number of
branch (96.2±28.76), number of root (8.4±0.078) and length of root (59.18±4.86
em) was found with cow dung 10 tonlha + N:P:K 45:30:15 kg/ha .This was
followed by cow dung 5 tonlha + N:P:K 60:40:20 kglha, but number of nodes
and internodes and number of branch was second highest in cow dung 5 tonfha +
N:P:K 45:30:15 kg/haas compared to control.
Estimation of Primary Metabolites:
Estimation of total sugar in cuttings of B. diffusa treated with different
combinations of fertilizer :
Maximum amount of total sugar with (15.46±0.088 mg/g, 14.2±0.174 mg/g
and 13.7±0.369 mg/g) in treated cutting was found with root, leaf and stem
respectively in cow dung 10 tonlha + N:P:K 45:30:15 kg/ha. The second highest
amount of total sugar was found with cow dung 5 tonlha + N:P:K 60:40:20 kg/ha
treated cutting, (14.56±0.086 mg/g, !3.93 ±0.384 mg/g and 13.16±1.92 mg/g), in
root leaf and stem respectively. Control had values (11.13±0.066 mg/g,
I 0.33±0.143 mg/g and I 0.66±0.142 mg/g) in root, leaf and stem respectively.
!02
Estimation of reducing sugar in cuttings of B. diffusa treated with
different combinations of fertilizer:
Maximum amount of reducing sugar (2. 7±0.1 mg/g) was found in root with
cow dung l 0 ton/ha + N:P:K 60:40:20 kg/ha treated but in leaf and stem
( 1.8±0.057) mg/g and ( 1.86±0.18 mg/g) but in root it was maximum in cow dung
10 ton/ha + N:P:K 45:30:15 kg/ha treated cuttings followed by (2.5±0.057 mg!g,
1.56±0.202 mg/g and 1.73±0.120 mg/g) in root, leaf and stem of cow dung 5
ton/ha + N:P:K 60:40:20 kg/ha respectively as compared to (1.73±0.120 mg/g,
1±0.152 mg/g and 1.26±0.03 mg/g) in root, leaf and stem of control.
Estimation of total soluble protein in cuttings of B. diffusa treated with
different combinations of fertilizer:
Maximum amount of total soluble protein (31.23±0.66 mg/g and
14.8±0.404 mg/g) was found with root and stem respectively of cow dung 10
tonlha + N:P:K 45:30:15 kg/ha but in leaf (18.0±0.14 mg/g) it was maximum in
cow dung 10 ton/ha N:P:K 30:20:10 kg/ha treated cutting.
Estimation of TCA precipitated protein in cuttings of B. diffusa treated
with different fertilizers combinations:
Maximum amount of TCA precipitated protein was (5.13±0.606 mg/g) in
root and (3.66±0.589 mg/g) and leaf in cow dung 10 tonlha N:P:K 45:30:15 kg/ha
treated cutting but in stem with (3.93±0.589 mg/g) it was maximum in cow dung
10 ton!ha + N:P:K 60:40:20 kg/ha treated cutting followed by (4.26±0.145 mg/g,
2.09±0.063 mg/g, 3.86±0.290 mg/g) in root, leaf and stem respectively in cow
dung 5 ton/ha + N:P:K 60:40:20 kg/ha treated cuttings as compared to (3.86±2.90
mg/g, 1.86±0.520 mg/g and 2.06±0.133 mg/g) in root, leaf and stem of control.
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Estimation of chlorophyll content in cuttings of B. diffusa treated with
different fertilizers combinations:
Maximum amount of chlorophyll a, chlorophyll b, total chlorophyll and
carotenoides were (0.663±0.023 mg/g, 0.346±0.002 mg/g, 0.991±0.029 mg/g and
0.516±0.008 mg/g) with cow dung 10 ton/ha + N:P:K 60:40:20 kg/haas compared
to (0.309±0.008 mg/g, 0.170±0.008 mg/g, 0.681±0.031 mg/g and 0.292±0.01 9
mg/g) in control respectively.
Amount of alkaloid in roots of B. dijfusa treated with different
fertilizers combinations:
The amount of alkaloid was increased from cow dung 0 ton/ha + N:P:K 0
kg/ha to cow dung 10 ton/ha + N:P:K 45:30:15 kg/ha treated cuttings. Maximum
amount of alkaloid was (0.0511) gm/gm was found with cow dung 10 ton/ha +
N:P:K 45:30:15 kg/ha treated cutting followed by cow dung 5 tonlha + N:P:K
60:40:20 kg/ha treated cutting was (0.0285 gm/gm) alkaloid as compared to
(0_.0185 gm/gm) in control.
DETERMINATION OF SUITABLE SOIL FOR SOLANUM
XANTHOCARPUM:
Plant grown in Kanhar soil gave highest value for almost all the growth
parameters as compared to plant grown in Bhata or Matasi soil. Maximum plant
length (13.24 ±3.147 em), number of leaves (17.98±4.594), size of leaves
(length& width) (6.09±3.263 em & 4.02±0.67 em), number of nodes and
internodes (12.9±3.977 and 11.9±2.81 ), number of branch (7.0±2.317), number of
flowers (5.9±3.26) and number of fruits (8.848±1.54) were obtained with Kanhar
soil followed by plant grown in Bhata and Matasi soil.
104
Estimation of Primat·y metabolites:
Maximum amount of total sugar was (10.42±0.037 mg/g and 9.46±0.185)
mg/g found in leaf and stem respectively of plant grown in Kanhar soil but in root
total sugar was maximum (7.366±0.233 mg/g) in plant grown in Bhata soil.
Estimation of reducing sugar in plant grown Kanhar, Bhata and Matasi
soil grown plants:
The amount of reducing sugar was maximum in root ( 1.82±0 .120 mg/ g)
m plant grown in Bhata soil and almost equal amount of reducing sugar
(1.16±0.06 mg/g and 1.1±0.2 mg/g) was found in root of plant grown in Kanhar
and Matasi soil. The amount of reducing sugar in leaf and stem of plant grown in
Kanhar soil were (1.53±0.033 mg/g).
Estimation of total soluble protein in plant grown in Kanhar, Bhata and
Matasi soil :
Maximum amount of total soluble protein found in leaf, stem and root of
plant grown in Kanhar soil were (15.83±0.233 mglg 10.13±0.410 mg/g and
6.9±0.272 mg/g) respectively followed by plant grown in Bhata soil (15.3±0.272
mg/g, 9.6±0.3.8 mg/g, 6.1±0.351 mg/g) and Matasi soil (14.9±0.655 mg/g
9.2±0.450 mg/g and 5.83 ± 0.176 mg/g) in leaf, stem and root respectively.
Estimation of TCA precipitated protein in plant grown Kanhar, Bhata
and Matasi soil :
Maximum amount of TCA precipitated protein (6.466±0.233 mg/g,
5.83±0.02 mg/g) and (2.46±0.202 mg/g) found in root, leaf and stem respectively
of plant grown in Kanhar soil, followed by plant grown in Bhata soil (6.03±0.123
mglg 2.8±0.491 mg/g, 2.33±0.246 mg/g) and Matasi soil (5.73±0.33 mg/g
3.4±0.33 mg/g and 1.66±0.881 mgfg) in root and leaf and stem respectively.
105
Estimation of chlorophyll content in plant grown in Kanbar, Bhata and
Matasi soil:
The maximum amount of chlorophyll a, chlorophyll b, total chlorophyll and
carotenoides were found with plant grown in Kanhar soil with (0.306±0.007 mg/g,
0.089±0.009 mgfg, 0.396±0.016mgfg and 0.272±0.10 mgfg) respectively followed
by values with plants grown in Bhata and Matasi soil.
Amount of alkaloid in berries of S. xanthocarpum grown in Kanhar,
Matasi and Bhata soil:
Maximum amount of alkaloid was (0.11 02 gm/gm) found in plant grown in
Kanhar soil followed by Bhata and Matasi soil were (0.978 grn/gm) and (0.0759
gm/gm) respectively.
HORMONE TREATMENT:
IBA treated cutting showed better response on plant growth as
height/length , number of leaves, size of leaves, number of nodes and internodes,
number of branch, number and length of root as compared to IAA treated cuttings
and control.
IBA 50ppm treated cuttings showed maximum plant height (36.57±2.72 em), but
number of leaves (15.44±5.23), size of leaves (length & width) (8.315±0.812 em
& 5.11±0.592cm), number of nodes and internodes (7.28±1.29 & 6.28±1.29),
number of branch (5.52±0.585), number of flower (6.52±0.733) and number of
fruit (5.08±1.05) and number of root (26.4±0.92) and length of root (14.16±0.756)
em was found with IBA 1 OOppm treated cuttings followed by value with lAA
50ppm treated cutting.
106
Estimation of Primary metabolites:
Estimation total sugar in cuttings of S. xantltocarpum treated with 10,
50 and lOOppm of IAA and IBA:
The amount of total sugar was maximum 18±0.435 mg/g, 13.8±0.688 mg/g
and 12.9±0.3~8 mg/g) was found in leaf, stem and root respectively of !OOppm
treated cuttings of IBA. Minimum amount of total sugar was found with IAA
lOppm treated cuttings. Control had values (9.533±0.328mg/g, 10.7±0.115 mg/g
and 10.1±0.1 mg/g) in leaf, stem and root respectively.
Estimation of reducing sugar in cuttings of S. xanthocarpum treated
with 10, 50 and 1 OOppm of IAA and IBA :
Maximum amount of reducing sugar was (1.8±0.043 mglg, 1.46±0.088
mg/g) found with leaf and stem respectively of IBA 1 OOppm treated cutting but in
root it was maximum (1.26±0.03 mg/g) in IAA 50ppm treated cuttings. This was
followed by IAA 1 OOppm treated cutting Minimum amount of reducing sugar was
found in leaf, stem and root respectively ofiAA 1 Oppm treated cuttings.
Estimation of total soluble protein in cuttings of S. xanthocarpum
treated with 10, 50 and lOOppm ofiAA and IBA:
Maximum amount of total soluble protein was (39.53±0.721 mglg,
23.2±0.527 mg/g and 39.36±0.435 mglg, 23.13±0.86 mg/g) was found with IBA
1 OOppm and IAA 50ppm treated cuttings of leaf and root respectively. But in stem
it was maximum (16.56±0.033 mg/g) in IBA 100ppm treated cuttings. Minimum
amount of total soluble protein (30.2±0.433 mg/g, 14.03±0.463 mg/g and
12.36±0.323 mg/g) in root leaf and stem of IAA lOppm treated cutting as
compared to (22.63±1.31 mg/g, 10.36±0.72 and I 1.9±0.176 mg/g) in leaf, stem
and root of control.
107
Estimation of total soluble protein in cuttings of S. xantlwcarpum
treated with I 0, 50 and lOOppm ofiAA and lBA :
Maximum amount of total soluble protein was (39.53±0.721 mglg,
23.2±0.527 mglg and 39.36±0.435 mg/g, 23.13±0.86 mg/g) was found with IBA
1 OOppm and IAA 50ppm treated cuttings of leaf and root respectively. But in stem
it was maximum ( 16.56±0.033 mg/g) in II3A 1 OOppm treated cuttings. Minimum
amount of total soluble protein (30.2±0.433 mg/g, 14.03±0.463 mg/g)and
12.36±0.323 mg/g) in root leaf and stem of IAA lOppm treated cutting as
compared to (22.63±1.31 mg/g, 10.36±0.72 and 11.9±0.176 mg/g) in leaf, stem
and root of control.
Estimation of TCA precipitated protein in cuttings of S. xanthocarpum
treated with different concentration 10, 50 and lOOppm of IAA and
IBA:
Maximum amount of TCA precipitated protein was (10.8±1.44mglg,
2.63±0. 753mg/g) in leaf and stem respectively of IBA 1 OOppm treated cutting but
in root it was maximum (3.6±0.264 mg/g) in IAA lOOppm treated cuttings. This
was followed by (9.83±0.207mg/g, 2.03±0.392 mg/g and 2.13±1.20 mg/g) in leaf,
stem and root respectively of IAA 50ppm treated cuttings. Minimum amount of
TCA precipitated protein was (6.03±0.120 mg/g, 2.06±0.881 mg/g and 1.26±0.03
mglg) was found in leaf stem and root ofiAA 1 Oppm treated cuttings. Control had
values (3.43±0.120 mg/g, 1.1±0.06 mg/g and 1.66±0.881 mg/g) in leaf stem and
root respectively.
Estimation of chlorophyll content in cuttings of S. xanthocarpum treated
with different concentration 10, 50 and lOOppm of IAA and IBA:
The maximum amount of chlorophyll a, chlorophyll b, total chlorophyll and
carotenoides in sequences of(0.389±0.005 mg/g, 0.221±0.031 mg/g, 0.267±0.133
mg/g, 0.541±0.035) mg/g was found with IAA lOppm treated cutting followed by
IBA I Oppm treated cutting. The minimum amount chlorophyll a, chlorophyll b,
108
total chlorophyll and carotenoides was found with IAA 50ppm treated cuttings as
compared to control.
Amount of alkaloid in bcnics of S. xanthocarpum treated with 10, 50
and 100 ppm ofiAA and IBA:
Maximum amount of alkaloid (0.5543 gm/gm)was found with IBA 50ppm
treated cutting followed by IAA 50 ppm treated cutting (0.05279 gm/gm) and
minimum amount of alkaloid was (0.2295 gm/gm) found with IB I 0 ppm treated
cuttings as compared to (0.1082gm/gm) in control.
FERTILIZER TREATMENT:
Maximum plant height (59.54±25.41 mg/g), number of leaves
(211.08±15.20 mg/g), size (length & width) (6.75±1.46 mg/g, 5.325±1.21 mglg),
number of nodes and internodes (86.29±49.28 mglg & 85.64±49.14 mglg),
number of t1ower (17.2±6.305 mg/g) and number of fruit (47.36±33.6 mglg) was
found with cow dung 10 tonlha + N:P:K 60:40:20 kglha treated cuttings but
number of branch was maximum 21.92±9.25 mg/g in cow dung 10 ton/ha + N:P:K
45:30:15 kg/ha, treated cuttings followed by cow dung 5 ton/ha + N:P:K 60:40:20
kglha.
Estimation of Primary Metabolites:
Maximum amount of total sugar with (14.33±1.78mglg, 9.33±1.03 mglg
and 14.7±5.1 mglg) in leaf, stem and root of cow dung 10 tonlha + N:P:K
60:40:20 kglha similar in cow dung 10 tonlha + N:P:K 45:35:15 kglha and cow
dung 10 ton/ha + N:P:K 30:20:10 kglha treated plants. The second highest amount
of total sugar (12.9±0.176 mglg, 7.66±0.466 mg/g andl3.1±0.115 mglg) was
found with leaf stem and root of cow dung 5 tonlha + N:P:K 45:35:15 kglha
treated cuttings as compared to (9.43±0.21 mglg, 5.36±0.676 and 1 0.36±0.120
mg/g) in kaf. st~m and root of control respectively.
109
Estimation of reducing sugar S. xantlrocarpum treated with different
combinations of fertilizer:
The maximum amount of reducing sugar (2.7±0.057 mg/g, 1.66±0.120
mg/g and 3.06 mg/g) in leaf, stem and root respectively of cow dung 10 tonlha +
N:P:K 60:40:20 kg/ha treated cuttings followed by 5 ton/ha + N:P:K 60:40:20
kg/ha treated cu!tings. Control had values (!. 73±0.08 mg/g, 1.13±0.03 mg/g and
1.46±0.317) mg/g in leaf, stem and root respectively.
Estimation of total soluble protein S. xanthocarpum treated with
different combinations of fertilizer:
Maximum amount of total soluble protein was (26.13±0.744 mg/g,
7.46±0.554 mg/g, 10.06± 0.133 mg/g) in leaf, stem and root respectively of cow
dung !0 tonlha + N:P:K 30:20:10 kg/ha treated cuttings followed by cow dung 5
tonlha + N:P:K 60:40:20 kg/ha treated cuttings was (23.93±0.920 mg/g, 7.33±0.06
mg/g and 8.4±0.723 mg/g) found with leaf, stem and root respectively as
compared to (!7.!6±0.833 mg/g, 4.56±0.554 mg/g and 6.3±0.05 mg/g) in leaf,
stem and root of respectively recorded for control.
Estimation of TCA precipitated protein S. xantlwcarpum treated with
different combinations of fertilizer:
Maximum amount of TCA precipitated protein was (10.96±0.491 mg/g,
5.83±0.317 mg/g and 6.56±0.463 mg/g) in leaf, stem and root respectively of cow
dung 10 ton/ha + N:P:K 30:20:10 kg/ha treated cutting. This was followed by 10
ton/ha + N:P:K 45:35:15 kg/ha (9.8±0.461 mg/g, 4.36±0.218 mg/g and 5.83±0.202
mg/g) respectively as compared to (5.96±0.554 mg/g, 2.13±0.57 mg/g) in leaf,
stem and root of recorded for control.
110
Estimation or chlorophyll a, chlorophyll b, total chlorophyll and
carotenoidcs in S. xant!tocarpum treated with different fertilizer
combinations:
Maximum amount of chlorophyll a, chlorophyll b, total chlorophyll and
carotcnoidcs \\as (0.74}J, 0.074 mg/g. 0.225±0.014 mg/g, 0.909±0.02 mg/g and
0.385±0.100 mg/g) was found with cow dung 10 ton/ha + N:P:K 60:40:20 kg/ha,
as compared to (0.324±0.013 mglg, 0.092±0.003 mg/g, 0.422±0.018 mg/g and
0.286±0.020 mg/g) respectively recorded for control.
Amount of alkaloid in berries of S. xanthocarpum treated with different
fertilizer combinations:
Amount of alkaloid in berries of S. xanthocarpum was increasing in cow
dung 0 ton!ha + N:P:K 0 kg/ha to cow dung 10 tonlba + N:P:K 60:40:20 kg/ha.
Maximum amount of alkaloid in berries was (0.7770 gm/gm) found in cow dung
10 ton/ha + N:P:K 60:40:20 kglha treated plant as compare to (0.1589 gm/gm) in
cow dung 0 ton/ha + N:P:K 0 kg/ha.
CONCLUSION
Based on the above findings the following major conclusions can be drawn:
•:• For determination of suitable soil three types of the soil Kahhar, Matasi and
Bhata soil were used. The Kanhar soil was the suitable soil for vegetative
growth of Boerhaavia diffusa.
•:• The maximum amount of total sugar was found in root, leaf and stem of
cuttings of B. dij]i1sa grown in Kanhar soil
•:• Maximum amount of reducing sugar was found in root and leaf
respectively of cuttings grown in Kanhar soil, but in stem reducing sugar
was maximum in cuttings grown in Matasi soil.
\\I
•!• Maximum amount of total soluble protein was found in root, leaf and stem
respectively in cuttings of B. dijfitsa grown in Kanhar soil.
•!• Maximum amount of TCA precipitated protein was found in root and leaf
of cuttings of B. dil}itsa grown in Kanhar soil but in stem it was maximum
in culting grown in Bhata soil.
•!• The amount of chlorophyll was found to be higher in cuttings of B. di/fitsa
grown in Kanhar soiL
•!• The amount of alkaloid in root of B. diffusa was also maximum in the
cuttings B. diffusa grown in Kanhar soil.
•!• Plant grown in Kanhar soil gave highest values for vegetative growth of S
xanthocarpum.
•!• Maximum amount of total sugar was found in leaf and stem of plant grown
in Kanhar soil but in root it was maximum plant grown in Bhata soil.
•!• Maximum amount of reducing sugar was found in leaf and stem of plant
grown in Kanhar soil but in root it was maximum plant grown in Bhata soil.
•!• The amount of total soluble protein and TCA precipitated protein were
maximum in plant grown in Kanhar soil.
•!• · The amount of alkaloid in berries of S. xanthocarpum was also maximum
in the plant grown in Kanhar soil.
•!• For hormone treatment, IAA and IBA in three concentrations l Oppm,
50ppm and IOOppm were used. 50ppm IBA showed better response for
vegetative growth of B. di/fi1sa and 1 OOppm IBA treated cutting showed
better response for vegetative growth of S. xanthocarpum.
•!• The amount of total sugar and reducing was maximum in leaf, stem and
root of the 50ppm IBA treated cuttings of B. diffusa.
•!• Maximum amount of total soluble protein and TCA precipitated protein
was maximum in leaf and root of SOppm IBA treated cuttings of B. diffusa
but in stem it was maximum in IOOppm IBA.
•!• The amount of chlorophyll was found to be higher in cuttings treated of B.
diffilsa treated with 50pprn IBA.
112
•!• The amount of alkaloid in root of B. diffusa was also maximum in the
cuttings tn:akd with 50ppm 18!\..
•!• Maximum amount of total sugar was found in leaf, stem and root of
cuttings of S. xanthocarpum treated with I OOppm IBA and reducing sugar
was maximum in leaf and stem of cutting treated with 1 OOppm IBA but in
root it was maximum in 50ppm!AA treated cuttings.
•!• Maximum amount of total soluble protein and TCA precipitated protein
was leaf and root of 1 OOppm IBA and 50ppm IAA treated cuttings of S.
xanthocarpwn but in stem it was maximum in 1 OOppm IBA
•!• The amount of chlorophyll was found to be higher in cuttings treated of S.
xanthocarpum treated with IOOppm IBA.
•!• The amount of alkaloid in berries of S. xanthocarpum was also maximum
in the cuttings treated with 1 OOppm IBA.
•!• For the effect of fertilizer treatment various combination as cow dung 10
ton/ha + N:P:K 60:40:20 kglha, cow dunglO ton/ha + N:P:K 45:30:15
kglha, cow dung 10 ton/ha + N:P:K 30:20:10 kglha, and cow dung 5 ton!ha
+ N:P:K 60:40:20 kg/ha, cow dung 5 ton/ha + N:P:K 45:30:15 kg/ha and
cow dung 5 ton/ha + N:P:K 30:20:10 kg!ha were applied in cuttings of B.
diffusa and plant of S. xanthocarpum. Cow dung 10 tonlha + N:P:K
45:30: I 5 kg/ha and cow dung 10 ton!ha + N:P:K 60:40:20 kg/ha gave better
response of vegetative growth in cuttings of B. diffusa and plant of S.
xanthocarpum.
•!• Total sugar was found maximum in leaf, stem and root of the cuttings of B.
diffusa treated with cow dung 10 ton/ha + N:P:K 45:30:15 kglha and
reducing sugar was found maximum in leaf and stem of cutting treated with
cow dung!O tonlha + N:P:K 45:30:15 kg!ha but root it was maximum in
cow dung 10 ton/ha + N:P:K 60:40:20 kg/ha.
•!• Total soluble prott:in was maximum in root and stem of B. diffusa found in
cow dung 10 tonlha + N:P:K 45:30:15 kg/ha but in leaf it was maximum in
cow dung 10 ton/ha + N:P:K 30:20:10 kg/ha and TCA precipitated protein
113
was maximum in root and leaf of B. diflusa found in cow dung 10 ton/ha +
N:P:K 45:30:15 kg/hand in stem it was maximum in cow dung 10 ton/ha +
N:P:K (10:40:20 kg/In
•!• Total sugar and reducing sugar was found maximum in leaf, stem and root
of the S. xamhocarpum treated with cow dung I 0 ton/ha + N:P:K 60:40:20
kg/ha.
•!• Maximum amount of total soluble protein and TCA precipitated protein
was found in root, leaf and stem of S. xanthocarpum were cow dung I 0
ton/ha + N:P:K 30:20:10 kg/ha.
•!• The maximum amount of chlorophyll content in leaf of B. diffusa and S.
xanthocarpum was found with cow dung I 0 ton/ha + N:P:K 60:40:20 kg/ha.
•!• The maximum amount of alkaloid in roots of B. diffusa found in cow dung
10 ton/ha + N:P:K 45:30:15 kg/ha and berries of S. xanthocarpum found m
cow dung 10 ton/ha + N:P:K 60:40:20 kglha.
