AN INVESIGATION ON THE DROUGHT STRESS TOLERANCE POTENTIAL

OF PHASEOLUS VULGARIS L. AND VIGNA RADIATA L.

Chinthu R V *1 and Mr. Shal chandran *2
*1
Department of Botany University of Kerala ,Kariavattom
*2
Asst.professor, Dept of Botany ,S N College Cherthala
Email:chinthukailas@gmail.com

Abstract has become essential to have information

Drought is one of the most severe problems in
worldwide agricultural production. Seed
germination is critical and governs the later
growth and yield of the plant. The drought
stress studies were carried out in two species of
Pea plants viz. PHASEOLUS VULGARIS L.
AND VIGNA RADIATA L. which make the food
more proteinaceous. In addition, PHASEOLUS
VULGARIS L. AND VIGNA RADIATA L. have
many medicinal properties.The in vitro drought
was induced in these plant seeds by using
different levels of polyethylene glycol (PEG)
i.e., 5%, 10%, 15% and control. Observations
on the seed germination parameters were made
after 48 hours of sowing. The seed germination
percentage of both Phaseolus vulgaris L and
Vigna radiata L. was not much affected upon
drought stress as compared to their control. But
the embryonic root growth was much affected
in the case of. Vigna radiata L. as compared to
Phaseolus vulgaris L.This showed the high
drought withstanding potential of Phaseolus
vulgaris L. The proline and total sugar content
was increased significantly in Phaseolus
vulgaris L. as compared to Vigna radiata L.
upon drought stressed condition. Thus the
result of this study denotes the drought
tolerance potential of Phaseolus vulgaris L.
Key words: Drought stress , PEG , Seed
germination , Proline , Sugar
As a result of various abiotic and biotic stresses
there is a heavy loss in crop reported as in various
vegetable crops, the scientific community is
always look out for stress tolerant crop species /
variants. These stress tolerant crop species are
generated through various techniques, of which
the biochemical approach plays a major role. So it
regarding the drought stress tolerance potential of
various Pea species. This will ensure that the
cultivation of these species will not turn out to be
a failure in the middle of the growing season and
thus can ensure productivity.
MATERIALS AND METHODS
Seeds of two pea plants viz., and Phaseolus
vulgaris and Vigna radiata were collected from
Cherthala. The seeds were sown in soil mixed with
cow dung. after 30 days the plantlets were
transferred to polythene bags containing sand, soil,
cow dung in a ratio of 1:2:1.
Induction of drought by using PEG
The different concentration of poly ethylene glycol
(0, 5%,10%,15%) were prepared. These medium
were poured in to appropriately labelled
petriplates. Pea seeds from each species (10 in
number) and controls were transferred to
corresponding concentrations of PEG in aseptic
condition on to a Whatman filter platform. After
24 hour of incubation the seedling were screened
for germination properties as well as for their
growth rate . Investigations on germination were
carried out with reference to the following
parameters.
Germination percentage and root growth
The germination percentage was calculated
using the formula.
Germination percentage
= No of seeds germinated X 100
Total no of seeds sawn
 Embryonic root growth was measured by
using a standard scale.
Induction of Drought
Drought was imposed to one set of 12 plants by
withholding irrigation where as the other set of 12
plants were kept as control.After acclimatization
to this condition for 10 days, the leaves were
excised and collected in labelled polythene bags
for various analysis.
Estimation of Total Sugar
Total sugar content was estimated by using the
method of Dubois et.al (1956). 0.5 g of tissue was
homogenized in 10 ml of 70% ethanol and
centrifuged at 10,000 rpm for 15 minutes. From
each sample 1 ml of the supernatant was taken in
to test tubes. To this 1ml of 5% phenol solution
was added followed by 5 ml con.H2SO4. The
reagent blank was also prepared by taking the
entire reagent except sample.1 ml ethanol was
taken as blank instead of sample. The mixture was
then kept at 30 ͦ c for 20 minutes. Read the color of
the mixture at 490nm using spectrophotometer.
Estimation of Proline
Estimation of proline content was according to
Bates method (1973). 1g. of leaf tissue was
weighed & homogenized in 3 ml of 3%
sulphosalicylic acid. The homogenate was then of
centrifuged at a speed of 10000 rpm for 15
minutes. The supernatant was saved & used for the
estimation.From each sample 1 ml. of the
supernatant was taken in to test tubes & then 1ml.
of glacial acetic acid & 1ml of ninhydrin was
added to it. The test tubes were then covered with
round marbles & further kept in boiling water bath
for 1h. To terminate the reaction, the tubes were
dipped in cold water. The reagent blank was also
prepared by taking the entire reagent except the
sample. Instead of sample 1ml. of 3%
sulphosalicylic acid was taken. 4 ml of toluene was
then added to each test tube &shaken well using a
vortex mixture.The red chromophore was
collected & the absorbance was measured against
a reagent blank at 520nm using a
spectrophotometer.
RESULTS AND DISCUSSION
Germination percentage and Root Length
In this investigation it was found that the seed
germination of Phaseolus vulgaris L. and Vigna
radiata L. is not much inhibited even at high
concentration of PEG (Table.1). The growth of
embryonic root was found to be decreased at 10 %
of PEG in the case of Phaseolus vulgaris but in the
case of Vigna radiate it was not inhibited even at
10 % of PEG.
Total sugar content
Total sugar s may help the plants to bring about the
osmotic adjustment (Hare et.al 1998).In this
present investigation, Phaseolus vulgaris L. has
showed greatest potential to accumulate sugars at
enhanced level as compared to Vigna radiata L.
When there was 62.2 % ( 2120 µg/g fr. wt )
increase in the total sugar content in Phaseolus
vulgaris L on subjecting to drought stress as
compared to their control plants, it was only 32%
(2120 µg/g fr .wt ) increase in the case of Vigna
radiata L.
Total proline content
. It is a compatible solute and its play an
important role in the osmoregulation plant when
plants are exposed to various abiotic stresses
(Bohnert et.al, 1995). It can also prevent
membrane damage and protein denaturation by
scavenging free radicals. Therefore increasing
proline content can help the plants to tolerate
salinity stress. In this present investigation
Phaseolus vulgaris was recorded to accumulate
proline at enhanced level as compared to Vigna
radiata (Table.3). Phaseolus vulgaris showed
59.5% (1018 µg / g Fresh weight) increase in the
proline content on subjecting to salinity stress as
compared to their control plants whereas in Vigna
radiata it was only 8% (998 µg / g Fresh weight).
Phaseolus vulgaris
Control 5% 10% 15%

Fig: 1(a). Effect of the concentrations of P E G Stress

on seed germination of Phaseolus vulgaris
Vigna radiata (Fig : 2(b) )
Vigna radiate
Control 5% 10% 15%

Control
Fig: 1(b). Effect of the concentrations of P E G
on seed germination of Vigna radiate
Phaseolus vulgaris (Fig : 2(a) )

Control
Stress
Germination percentage
Table 1 Total sugar content
Table 3
Phaseolus Vigna
Concentration of vulgaris radiata
P E G (%) Total sugar content
(µg / g Fresh weight)
0% 100 100 Name of the Percentage
plant of difference
Drought
control
5% 100 100

Phaseolus 800 µg 2120 µg 62.2%

10% 100 100
vulgaris

15% 100 100

Vigna 1400 µg 2120µg 32%
radiata

Root growth
Total proline content
Table 2
Table 4
Proline content
Root length (cm)
Name of the (µg / g Fresh % of
5% 10% 5% plant weight) difference
Plants
Control PEG PEG P Drought
EG Control

Phaseolus 24.2 24.5 9 3 Phaseolus 412 1018 59.5

vulgaris
vulgaris

Vigna radiate 917 998 8

Vigna radiata 7.8 7.3 7.5 4
CONCLUSION
In the present study various parameters
have been made to analyze the drought tolerance
potential of Phaseolus vulgaris L. and Vigna
radiata L. and to compare its drought tolerance
capacities. For this purpose experiments to find
out the germination capabilities of these two
species at different concentration of P E G
(0%,5%,10% 15%) and various non enzymatic
mechanisms (proline and total sugar content) of
drought tolerance under water deficit condition.
The seed germination percentage of
both Phaseolus vulgaris L and Vigna radiata L.
was not much affected upon drought stress as
compared to their control. But the embryonic root
growth was much affected in the case of. Vigna
radiata L. as compared to Phaseolus vulgaris L.
This showed the high drought withstanding
potential of Phaseolus vulgaris L. The proline and
total sugar content were increased significantly
(59.5 & 62.2% respectively) in Phaseolus
vulgaris L .under drought stressesed condition as
compared to Vigna radiata L. These results again
pointed out the drought tolerance potential of
Phaseolus vulgaris L. and this data is very much
correlated with the result of root length studies in
seed.
From this present study it was well clear
that Phaseolus vulgaris L .is a drought tolerant
species in comparison with Vigna radiata L. Its
salt tolerance nature has been characterized by its
ability to maintain the embryonic root growth at
different concentration of PEG and efficient
osmoregulation capabilities (accumulation of
proline and total sugars).
In general the result of this study indicates the
higher drought tolerance potential of Phaseolus
vulgaris L., as compared to Vigna radiata L., and
it can be recommended for cultivation in drought
affected area.
ACKNOWLEDGEMENT
I expressed my sincere gratitude to Mr. Shal
Chandran, Asst.Professor Department of
Botany,all my teachers in the Department of
Botany, S N College Cherthala. Above all I am
thankful to the God almighty for the blessings
showered upon me.
LITERATURE CITED
1. An- Lhout F , Zunzunegui FA , Diaz Barrads
MC, Tinado R , Clavijio A ,Garcia Novo F (2001)
Comparison of proline accumulation in two
Meditteranean shrubs subjected to natural and
experimental water deficit.Plant soil 230:175-183.
2. Asada AK (1984) Chloroplast formation of
active oxygen and its scavenging. Methods
Enzymol 105: 422-428.
3. Ali M.L, M.S. Pathoo, J. Zhang , G. Bai, S.
Sarkarung & H.T. Nguyen 2000, Mapping QTL
for root traits in a recombinant inbreed population
from 2 indica ecotypes in rice Theor. Appl.Genet.
101 :756- 766.
4.Bird RP, Drapper HH(1984) Comparitive
studies on different methods of MDA
determination. Methods Enzymol 105: 299 - 305
5. Chandrasekhara RP, Vijnanabhaiah SW (1993)
Drought induced lipid peroxidation ; defencive
mechanism in upland rice (Oryza sativa) seeds
during germination. Adv plant Sci 6: 229 - 230.
6. Chiang HH, Dandekar AM (1995) Regulation
of proline accumulation in Arabiodopsis thaliana
(L). Heynh; during development and in response to
desication. Plant cell Environ 18: 1280 -1290.
7. Corcuera LT , Hintz M , Pahlich E (1989) Eff
ect of polyethylene glycol on protein extraction
and enzyme activation in potato cell cultures.
Phytochem 28: 569 - 1591.
8. Dimersevska K, Simova – Stoilova, Vassileava
V, Vaseva I , Grigorova B, Feller U (2008)
Drought induced leaf protein alteration in sensitive
and tolerant wheat variants. Gen. Appli. Plant
physiol 34(1 - 2).

9, Hare PD, Cress WA (1997) Metabolic

implications of stress induced proline
accumulation in plants. Plant growth Regul 21: 79
-102.

10. Hare PD, Cress WA Van staden J (1998)

Directing the roles of osmolyte accumulation
during stress.Plant Cell Environ 21: 535 - 553.

11. Ingram J , Bartel D (1996) The molecular basis

of dehydtation tolerence in plants. Annu Rev Plant
Physiol Plant Mol Biol 47: 377 - 403.

12.Lawlor DW, Cornic G (2002) Photosynthetic

carbon assimilation and associated metabolism in
relation to water deficit in higher plants. Plant Cell
Environ 25: 275 - 294.

13. Matysik J, Alia , Bhalu B, Mohanty P (2002)

Molecular mechanism of quenching of reactive
oxygen species by proline under stress in plants.
Curr Sci 82: 303 - 315.