National conference on "Conservation, Cultivation and Utilization of medicinal and Aromatic plants" (College of Horticulture, Mudigere Karnataka, 2018) Seed propagation studies in Embelia ribes burm. F

Abstract

Embelia ribes, commonly known as Vidanga or false pepper is a commercially important threatened medicinal plant of the Tropics belonging to the family Myrsinaceae which yields embelin, a highly valuable quinine derivative. Regeneration of the crop is poor due to over exploitation, diminishing forest area, dormant seeds, abortive embryos and hard seed coat. Seeds were subjected to several pre-soaking treatments i.e, soaking in water, aqueous solutions of GA3 at different concentrations, acid scarification using HCl, H2 SO4 and combination of acid scarification and GA3. Seeds treated with GA3 750 ppm for 24 hours and H2SO4 scarified seeds treated with GA3 750 ppm for 12 hours exhibited superior results compared to the untreated control. The shoot length (8.82 cm), root length (8.03 cm), and total dry biomass (70.21mg) were maximum in seeds treated with GA3 750 ppm whereas, germination percentage (87.5) and seedling vigour (1410.42) were maximum in H2SO4 scarified seeds treated with with GA3 750 ppm.

Full text

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Journal of Pharmacognosy and Phytochemistry 2018; SP3: 380-383
E-ISSN: 2278-4136
P-ISSN: 2349-8234
JPP 2018; SP3: 380-383
Aparna PM
College of Horticulture,
Bengaluru, Karnataka, India
Suryanarayana MA
ICAR- Indian Institute of
Horticultural Research,
Bengaluru, Karnataka, India
Rajasekharan PE
ICAR- Indian Institute of
Horticultural Research,
Bengaluru, Karnataka, India
Bhanuprakash K
ICAR- Central Plantation Crops
Research Institute, Kasaragod,
Kerala, India
Umesha K
College of Horticulture,
Bengaluru, Karnataka, India
Maruthi Prasad BN
College of Horticulture,
Bengaluru, Karnataka, India
Correspondence
Aparna PM
College of Horticulture,
Bengaluru, Karnataka, India
National conference on “Conservation, Cultivation and
Utilization of medicinal and Aromatic plants"
(College of Horticulture, Mudigere Karnataka, 2018)
Seed propagation studies in Embelia ribes burm. F.
Aparna PM, Suryanarayana MA, Rajasekharan PE, Bhanuprakash K,
Umesha K and Maruthi Prasad BN
Abstract
Embelia ribes, commonly known as Vidanga or false pepper is a commercially important threatened
medicinal plant of the Tropics belonging to the family Myrsinaceae which yields embelin, a highly
valuable quinine derivative. Regeneration of the crop is poor due to over exploitation, diminishing forest
area, dormant seeds, abortive embryos and hard seed coat. Seeds were subjected to several pre-soaking
treatments i.e, soaking in water, aqueous solutions of GA3 at different concentrations, acid scarification
using HCl, H2 SO4 and combination of acid scarification and GA3. Seeds treated with GA3 750 ppm for
24 hours and H2SO4 scarified seeds treated with GA3 750 ppm for 12 hours exhibited superior results
compared to the untreated control. The shoot length (8.82 cm), root length (8.03 cm), and total dry
biomass (70.21mg) were maximum in seeds treated with GA3 750 ppm whereas, germination percentage
(87.5) and seedling vigour (1410.42) were maximum in H2SO4 scarified seeds treated with with GA3 750
ppm.
Keywords: Embelia ribes, pre-sowing treatments, GA3, HCl, H2SO4
Introduction
Embelia ribes is a red listed, large woody climbing shrub belonging to the family Myrsinaceae
[10]. It is one of the 32 medicinal plants selected by National Medicinal Plants Board for its
large scale cultivation due to its commercial value [16-17]. It is mainly distributed in warmer
regions of northern and southern hemisphere. Most of the genera and species are tropical.
Genus Embelia, represented by more than 100 species, is distributed in Tropical Asia, Africa
and Australia and Pacific islands. The fruits, leaves, bark and root of Embelia ribes is of
immense value to the Traditional systems of medicine. It is an antipyretic, anticonvulsant,
antibacterial, antioestrogenic, antihelmintic, carminative, laxative, diuretic, and astringent [9]. It
is used in treatment of skin fungal infections, leprosy, hemorrhoids, obesity, lung diseases,
cancer, mental disorders and heart diseases [8, 2, 13]. The benzoquinone embelin obtained from
berries have proven antispermatogenic effects. Embelia ribes is one such plant which is
overexploited for commercial purpose. The threats faced by forest from humanshas eventually
led to the threatened status of medicinal plants like Vidanga. Regeneration of E. ribes from
seeds is poor, embryos are very small when present and most of the seeds are abortive. For the
survival and growth of E. ribes specific habitat conditions are essential. The propagation and
conservation of this important medicinal plant requires special attention.
Materials and Methods
Seed germination studies were conducted at Division of Plant Physiology and Biochemistry
and Division of Plant Genetic Resources, ICAR- Indian Institute of Horticultural Research,
Hessaraghatta, and Bengaluru. The experiment was carried out in the Seed germination
Chamber where the day and night temperature were maintained at an average of 20 0C /30 0C.
The required seed materials were collected from the plants grown in the Field Gene Bank of
RET medicinal plants maintained at Division of Plant Genetic Resources, ICAR- IIHR,
Bengaluru. The fruits were subjected to floatation test, where the sinkers were collected and
floaters were discarded. Then the mucilaginous seed coat present in the seeds was removed.
The seeds were treated with mercuric chloride 0.1 per cent for 10 minutes, later washed with
water and shade dried for 24 hours. The extracted seeds were subjected to 16 different pre-
sowing treatments. Treated seeds were sown in protrays consisting of coir pith and kept in

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Journal of Pharmacognosy and Phytochemistry
seed germination chamber (alternate temperature of 200C
/300C, 80-85% humidity). The protrays were covered with
black polythene films to maintain high humidity and
temperature to facilitate seed germination.
Table 1: Pre-sowing treatments given to Embelia ribes to enhance seed germination.
T1
Soaking in water
T2
GA3 at 250 ppm (12 h)
T3
GA3 at 500 ppm (12 h)
T4
GA3 at 750 ppm (12 h)
T5
GA3 at 250 ppm (24 h)
T6
GA3 at 500 ppm (24 h)
T7
GA3 at 750 ppm (24 h)
T8
HCl 35% (1 min )
T9
HCl 35% (1 min) + GA3 250 ppm ( 12 h)
T10
HCl 35% (1 min) + GA3 500 ppm ( 12 h)
T11
HCl 35% (1 min) + GA3 750 ppm ( 12 h)
T12
H2SO4 10% (10 min)
T13
H2SO4 10% (10 min) + GA3 250 ppm ( 12 h)
T14
H2SO4 10% (10 min) + GA3 500 ppm ( 12 h)
T15
H2SO4 10% (10 min) + GA3 750 ppm ( 12 h)
T16
Control
Fig1: a) Fruiting branch b) dried seeds c) ripe berries
Result and Discussion
The data pertaining to various germination parameters as
influenced by different pre-sowing treatments is summarized
in Table 2.
In this study the highest germination percentage (87.50) and
least number of days taken for initiation of germination (26)
was recorded in H2SO4 scarified seeds treated with GA3 500
ppm and H2SO4 scarified seeds treated with GA3 750 ppm
followed by seeds treated with GA3 750 ppm. The seedling
vigour was found to be highest in H2SO4 scarified seeds
treated with GA3 750 ppm (1410.42) and similar to that of
GA3 750 ppm (1372.70) followed by H2SO4 scarified seeds
treated with GA3 500 ppm. Other parameters like shoot
length, root length and total dry biomass of seedlings were
higher in seeds treated with GA3 750 ppm followed by
H2SO4 scarified seeds treated with GA3 750 ppm and
H2SO4 scarified seeds treated with GA3 500 ppm.
Table 2: Effect of preconditioning of seeds on days taken for initiation of germination and germination percentage.
Treatments
Days taken for initiation
of germination
Germination
percentage
Shoot length
90 DAS (cm)
Root length
90 DAS (cm)
Seedling
vigour
Total dry
biomass (mg)
T1- Soaking in water
50.00
30.50 (33.52)
4.84
2.64
228.04
39.28
T2- GA3 at 250 ppm (12 h)
43.00
42.50 (40.68)
5.70
3.18
377.73
40.37
T3- GA3 at 500 ppm (12 h)
41.75
44.00 (41.54)
5.90
3.36
407.33
43.63
T4- GA3 at 750 ppm (12 h)
40.00
47.50 (43.56)
6.23
3.55
464.45
45.16
T5- GA3 at 250 ppm (24 h)
33.25
60.50 (51.06)
7.50
6.17
827.02
56.07
T6- GA3 at 500 ppm (24 h)
31.50
78.00 (62.03)
7.68
7.55
1187.66
58.75
T7- GA3 at 750 ppm (24 h)
30.50
81.50 (64.53)
8.82
8.03
1372.70
70.21
T8- HCl 35% (1 min )
46.25
46.50 (42.99)
5.18
3.26
392.47
40.45
T9- HCl 35% (1 min) + GA3 250 ppm ( 12 h)
42.00
47.00 (43.27)
6.32
3.27
450.69
46.56
T10- HCl 35% (1 min) + GA3 500 ppm ( 12 h)
40.25
50.00 (45.00)
6.52
3.55
504.00
49.48
T11- HCl 35% (1 min) + GA3 750 ppm ( 12 h)
40.50
52.50 (46.43)
6.70
3.67
545.43
53.15
T12- H2SO4 10% (10 min)
41.25
50.50 (45.28)
5.49
3.49
453.53
42.26
T13- H2SO4 10% (10 min) + GA3 250 ppm ( 12 h)
31.00
73.00 (58.73)
8.05
6.73
1080.99
57.75
T14- H2SO4 10% (10 min) + GA3 500 ppm ( 12 h)
26.00
85.00 (67.24)
8.34
7.47
1345.74
59.44
T15- H2SO4 10% (10 min) + GA3 750 ppm ( 12 h)
28.00
87.50 (69.30)
8.41
7.61
1410.42
63.65
T16- Control
53.00
29.50 (32.88)
3.92
2.02
175.67
38.14
S.Em ±
0.75
0.73
0.10
0.08
18.37
0.66
CD @ 5%
2.13
1.73
0.30
0.25
52.25
1.89
* Values in the parentheses are arcsine transformed value

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Journal of Pharmacognosy and Phytochemistry
Fig 2: Effect of growth regulators and chemicals on days taken for
initiation of germination in E. ribes Burm. f. seeds
Fig 3: Effect of growth regulators and chemicals on shoot length and
root length in E. ribes Burm. f. seedlings at 90 days after sowing
Similar results were obtained in Embelia ribes where seeds
treated with GA3 500 ppm for 16 hours resulted in 80-85%
germination in 6-7 days [1, 18]. Reported that E. tsjeriam-
cottam seeds treated with 300 ppm GA3 recorded maximum
germination (52%) compared to control (13.20%). Seeds of
Embelia ribes when treated with GA3 500 ppm for 18 hours
recorded a germination percentage of 73.33 [5]. Treatment of
Vidanga seeds with GA3 750 ppm exhibited similar results in
case of early seed germination, germination rate, seedling
vigour, seedling height and number of leaves [19]. Gowda et al.
(2003) [6] reported that GA 400 ppm considerably improved
germination (48%) than control (12%) in E. tsjeriam-cottam.
Lavandula dentate seedstreated with gibberellic acid at 1000
ppm marked a maximum germination of 67% compared to the
control which did not exceed 1 per cent [3].
Among the different germination inducing treatments, the
seeds treated with gibberellins responded well with high seed
germination and vigorous seedling growth. Initiation of
germination was also earlier in GA3 treatment at different
concentrations. Paleg (1960) reported that, gibberellic acid
originating from the embryo is responsible for the hydrolysis
of starch reserves in the endosperm during germination of
grains. GA3 induces the de-novo synthesis of proteolytic
enzymes like α-amylase and ribonuclease. Amylases in turn
hydrolyse starch in the endosperm, providing the essential
sugars for the initiation of growth processes [4].
Low germination percentage in seeds may be due to physical
or chemical barriers like hard seed coat and dormancy. When
both these factors act together it’s a tough job for water to
penetrate and trigger germination. Acid scarification followed
by treatment with growth regulators like GA3 can easily solve
this problem. Here, highly appreciable results were obtained
in less time compared to that of GA3 treatment alone. Acid
treatment brings about softening of hard seed coat by
dissolution of pectic substances, lipids and high density
waxes, which is a common cause for hard seededness [7, 18].
This softens the seed coat in and makes it permeable to water
and gases. Futher, the GA3 treatment induces hydrolysis of
starch reserves which leads to germination of seed. Vidanga
seeds scarified with H2SO4 10% and then treated with GA3
500 ppm recorded the least number of days taken for
germination (26.00) with better germination (85%) whereas,
highest germination (87.5 %) and seedling vigour (1410.42)
was observed in H2SO4 scarified seeds treated with GA3 750
ppm.
Patwardhan et al. (2014) [12] reported that pre-sowing
treatment of 10 percent H2SO4 for 10 minutes + GA3 4000
ppm is the best treatment for Embelia ribes with 40%
germination. Pipinis et al. (2011) [14] observed higher
germination percentage (90.83) in Paliurus spina-christi Mill.
Seeds when scarified with H2SO4 (90 min) and then treated
with GA3 2000 ppm. Acid scarification considerably
increased the germination from 29.50 percent in control to 50
per cent. There was improvement in seedling parameters such
as total dry biomass (42.26 mg) and seedling vigour (453.53).
The seeds of Zanthoxylum armatum DC when treated with
diluted (50 %) H2SO4 (15 min) resulted in maximum
germination (93.3 %) along with mean germination time
(MGT) of 149.5 days [15]. Sharma et al. (2011) [18] reported
better germination (34%), rate of germination (0.95) and
vigour index (357) in E. tsjeriam-cottam seeds when treated
with concentrated H2SO4 for one minute. Though the seeds
soaked in water alone did not show appreciable results in this
experiment it might have aided the germination process along
with acids and GA3 by converting the insoluble food into
soluble form for its translocation to the embryo and by
bringing dissolved oxygen. We know that seedling emergence
is primarily a function of moisture availability and optimum
temperature to the seed. The tropical species require high
temperature and relative humidity for their germination [7].
This was achieved by covering the protrays with black
polythene. The seed germination chamber was a boon for the
seedlings from the harsh summer condition. From this
experiment it is evident that Vidanga seeds treated with GA3
750 ppm for 24 hours and H2SO4 scarified seeds treated with
with GA3 750 ppm for 12 hours exhibited nearly similar
results superior to the other treatments and control. The first
one reduces the cost and probable hazards from acids, but
requires long duration of treatment (24 hours) meanwhile, the
second one reduces the time required for seed treatment by 12
hours and slightly increases the cost involved in purchase of
chemicals.
Fig 4: Seedlings raised from the best treatments 90 days after sowing
a) GA3 500 ppm b) H2SO4 + GA3 500 ppm
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