African Journal of Biotechnology Vol. 7 (5), pp. 613-616, 4 March, 2008
Available online at http://www.academicjournals.org/AJB
ISSN 1684–5315 © 2008 Academic Journals
Ful Length Research Paper
Effect of phytoprotein treatment on Jatropha curcas for
Mousumi Debnath1* and H. N. Verma2
1Plant Biotechnology Laboratory, Department of Biotechnology, JECRC Foundation, Mahatma Gandhi Institute of
Applied Sciences, Tonk road, Jaipur, India -303905.
2Institute of Biotechnology and al ied sciences, Seedling Academy of Design, Technology and Management, Jagatpura,
Accepted 31 January, 2008
Jatropha curcas (Euphorbiaceae) is a perennial oil yielding plant. Jatropha is also host to a large
number of pathogens such as Cercospora sp. The systemic resistance and overall performance was
enhanced by seed soak treatment with leaf extract of Clerodendrum aculeatum. On treatment, the
treated plants not only showed systemic resistance but also showed marked enhancement in the plant
vegetative growth, rooting, flowering, fruiting and seed formation. Mechanisms underlying this may due
the synthesis of a new virus inhibitory phytoprotein produced in cells perceiving signals from systemic
resistance inducers or ribosome-inactivating functions of these antiviral proteins. These treated
Jatropha curcas can prove to be a miracle plant by turning wasteland into a money making land.
Key words: Jatropha curcas, biodiesel, phytoprotein , Clerodendrum aculeatum.
For a country reeling under the burden of a large oil
growth and seed yield. The plant has profuse vegetative
import bil and spiraling oil prices biodiesel is a promising
growth but the number of seeds produced per plant is
indigenous and renewable source of energy. Production
very low. Besides the plants produces seeds after
of bio-fuel from plant materials is a major step toward
approximately 2 - 3 years depending on the environmen-
harnessing renewable energy resources. Jatropha curcas
tal conditions. Seeds have limited viability and the plant is
(Euphorbiaceae) is a drought resistant, perennial plant
also prone to pathogens like Cercospora, Rhizopus
yielding 5 - 12 tones per hectares oil seeds and produces
oryzae, and insects.
2 - 4 tones of biodiesel. It may also transform the poor-
In spite of al these properties research on cultivation
est people and the most marginalized land into the
and propagation of J. curcas is limited. Thus it was
source for this energy (Kochar et al., 2005). It is a multi-
considered useful to undertake a systematic study and to
purpose tree with a long history of cultivation in the
develop a management strategy so that losses can be
tropical and subtropical regions of the world. It is a native
minimized. Phytoproteins derived from a few non hosts
of he central America and occurs mainly at low altitudes
healthy plants have recently shown to have potential use
in areas with annual temperature of wel above 30°C.The
as biocontrol agents (Baranwal and Verma, 2000). In
seeds are toxic due to the presence of curative ingre-
Ayurvedic system of medicine, certain plants and their
dients but after the treatment the seeds or the seed
extracts have been used to control viral diseases of
cakes can be used as animal feed. Jatropha is grown as
human beings and a number of plan products have been
a boundary fence to protect field from the grazing animals
identified through photochemistry (Sukh Dev, 2006).
and as a hedge to prevent erosion. The problem of great
These agents appear to act by sensitizing the plant for
concern regarding the plant is the rate of vegetative
activation of resistance mechanism. The leaves of
Clerodendrum aculeatum (Verbenaceae) and roots of
Boerhaavia diffusa (Nyctaginaceae) have been shown to
contain potent endogenous virus inhibitory proteins cal ed
the BD-SRI and CA-SRI (Awasthi and Verma, 2006).
*Corresponding author. E-mail: firstname.lastname@example.org.
These phytoproteins confer strong systemic resistance in
614 Afr. J. Biotechnol.
Table 1. Effect of phytoprotein treatment in J. curcas.
1. Height (cm)
2. Leaf: stem
0.79 ± 0.11
1.14 ± 0.15
a. Number of branches of stems
12 ± 2.04
29 ± 2.55
b. Diameter of the stem (cm)
a. Length (cm)
15 ± 0.10
5 ± 0.11
39 ± 1.26
20 ± 1.35
10 ± 0.86
a. size (in cm)
3.1 ± 0.0.1
3.4 ± 0.0.1
b. number per branch
10 ± 0.008
6 ± 0.01
a. Size (in cm)
b. Oil content (%)
30 ± 0.11
32 ± 0.25
Each value represents the mean ±SD, n = 10.
several plants against a large number of plant viruses
in the treated plants (Table 1). After 3 months there was
(Srivastava et al., 2004) within 4 - 6 h and at the same
vigorous rate of vegetative growth with increased branch-
time enhance vegetative growth. These phytoproteins
ing and overal better growth of the plants in comparison
have a molecular mass of 30 and 34 KDa respectively.
to the control, untreated plants (Figure 1A, B). Cutting of
The phytoproteins are highly stable and could be purified.
these plants can be a viable source of vegetative propa-
This can be a better approach towards introducing
gation. From the treated plants more than 20 - 25 such
resistance against virus in crops and simultaneously
propagules for cutting can be easily obtained for macro-
al ow these antiviral compounds to act as bioenhancers
propagation. The height, stem diameter and number of
and increase their plant growth simultaneously (Verma et
leaves were also more in comparison to the control
al., 1996). The present investigations were carried out to
plants. The number of the leaves and leaf: stem ratio was
study the effect of seed soak treatment of J. curcas with
also evaluated and found to be more in comparison to the
the antiviral resistance inducing phytoprotein extracted
control plants (Figure 1C) .These treated plants can be
from leaves of Clerodendron aculeatum (Verbenaceae).
thus useful as planting material for the vegetative
The roots also showed profound differences in growth.
MATERIALS AND METHODS
The treated J. curcas plants were deep rooted and
penetrating in comparison to the control plants. When the
During the present study the seeds of J. curcas were soaked and
root and rootlet length of the tap root was compare,
given an overnight pretreatment in 12% C. aculeatum plant extract.
marked differences were observed. The treated plants
One set of seeds was also soaked in plain distil ed water and kept
showed very long, thin (0.5 – 5 cm) roots that were exten-
as control. The overal growth, vegetative, rooting, flowering, fruiting
and seed formation were recorded at frequent intervals and
dable upto 60 cm in comparison to the control plants
compared with control plants. The oil content of the dried, crushed
having short (15 cm) and thick (2 – 3 cm) roots after 3
seeds of treated and the control plants were estimated using
months of growth. The roots gave rise to profuse rootlets.
petroleum ether as solvent by Electronic SOCS PLUS Automatic
The rootlets in the treated plants were many in number
Solvent Extraction System.
with further branching. In the treated plants the branches
was longer as wel as more in number (Figure 1D). The
treated plants with their deep penetrating roots can be
RESULTS AND DISCUSSION
used for soil reclamation as wel as phytoremediation of
various metal ions and minerals. This observation led to
In the present study there was better faster germination
the conclusion that the root characteristics of the treated
rate in the treated seeds as compared to the control.
and the control plants had marked differences in their
There was a rapid increase in the vegetative growth rate
morphological and functional abilities.
Debnath and Verma 615
Figure 1. A. Vegetative growth of Jatropha curcas control plants after 3 months of growth. B.
Vegetative growth of treated plants after 3 months of growth. C. Enhanced branching and height of
treated in comparison to control plants. D. Deep rooting with enhanced branching in roots and rootlets
of treated (left) in comparison to control (right) plants. E. More flowers in control plants. F. More female
flowers in treated plants. G. Fruits are more in number and smal in size in control plants. H. Fruits are
less in number and bigger in size in treated plants. I. Fruits of control plants in various stages. J. Fruits
of treated plants in various stages. K. The fruits and seeds after first yield. L. Seeds of control plants.
M. Seeds of treated plants.
616 Afr. J. Biotechnol.
Early flowering was observed in the plants. Flowering
This is the 1st report of seed soak treatment of J.
was observed after 9 months in the treated plants. There
curcas with 12% leaves extract of C. aculeatum. The
were more number of flowers in the control plants (Figure
development of disease resistant variety itself can
1E) as compared to the phytoprotein treated plants. In
provide the best option but breeding disease resistance
the 1st flowering phase there was maximum number of
varieties is tedious and time consuming. The enhanced
flowers in the control plants but al the flowers withered
growth by the activity of these bioenhancers was reflect-
and resulted in no fruit set in both control and treated
ed by substantial increase in height, leaf area, fresh
plants. In the 2nd flowering phase, 10 months after sow-
shoot weight, fresh root weight, root length, increased
ing, there was enhanced flowering in both control and
number of flowers, fruits, seeds, oil content and better
treated plants. The number of female flowers was slightly
resistance towards the development of systemic disease
increased in the treated plants (Figure 1F) and the fruit
symptoms of viral infections. The overal crop
characteristics were also recorded on the 11th month. The
performance was satisfactory and these treated plants
numbers of the fruits in a single branch were more in
were least affected by biotic and abiotic factors like
number in the control plants as compared to the treated
rainfal , frost and infes-tation by insects and diseases.
plants (Figure 1G, H). But when the size of the fruits was
These treated J. curcas can prove to be a miracle plant
correlated it was found that there was increased fruit size
by turning wasteland into a money making land.
and numbers of seeds were also more in treated plants.
The numbers of fruits in the treated plants were bigger in
size (Figure 1I, J and K). The seeds were also bigger in
size in the treated plants (Figure 1 L and M). The oil
Awasthi LP, Kumar P (2003). Protection of some cucurbitaceous crops
content of the seeds of treated and the control plants
against natural infection of viruses through Boerhaavia diffusa plants.
were evaluated and a 2% increase in the treated plants
Indian Phytopathol. 53: 317-319.
was estimated.Thus it was concluded that the soak
Awasthi LP, Verma HN (2006). Boerhaavia diffusa – A wild herb with
potent biological and antimicrobial properties. Asian–Agrihistory 10:
treatment of the leaves of C. aculeatum resulted in
miraculous increase not only in vegetative stages
Baranwal VK, Verma HN (2000). Antiviral phytoproteins as biocontrol
(branching of the shoots, deep penetrating roots) as wel
agents for efficient management of plant virus diseases. In:
as in the reproductive stages (early flowering, increase in
Biocontrol potential and its exploitation in sustainable agriculture
(Eds. Upadhay RK, Mukherjee KG, Chamola BP). Kluwer Academic/
female flowers, increase in fruit and seed size and oil
Plenum Publishers, New York, pp. 71-79.
content). These results clearly indicate that the plants on
Gupta RK, Srivastava A, Verma HN (2004). Cal us Culture and
seed soak treatment are not only resistant to many
Organogenesis in Boerhaavia diffusa : A Potent Antiviral Protein
pathogens but also show improved growth characte-
containing Plant. Physiol. Mol. Biol. Plants 10: 263-268.
Kochar S, Kochhar VK, Singh SP, Thind BS (2005). Differential rooting
ristics. Several reports are available on the effectiveness
and sprouting behavior of two Jatropha species and associated
of the abiotic agents as elicitors of resistance in
physiological and biochemical changes. Curr. Sci. 89: 936-938.
susceptible plant host. These agents appear to act by
Srivastava A, Gupta RK, Verma HN (2004). Micropropagation of
sensitisizing the plants for activation of resistance
Clerodendrum aculeatum through adventitious shoot induction and
production of consistent amount of virus resistance inducing protein.
mechanism (Gupta et al., 2004; Srivastava et al., 2004).
Indian J.Exp. Biol. 42: 1200-1207.
Such plant substances are known as systemic resistance
Sukh Dev (2006). A selection of prime ayurvedic plant drugs. Ancient-
inducers (SRI). The induction of the resistance is
Modern concordance, Anamaya publishers, New Delhi pp. 108-112.
expressed as reduction in lesion number in host reacting
Verma A, Verma HN (1993) Management of viral diseases of
mongbean by Clerodendrum aculeatum leaf extracts. Indian J. Plant
hypersensitivity to viral infection. Til now these SRI were
Pathol. 11: 63-65.
sprayed after every week (Verma and Verma, 1993;
Verma HN, Srivastava S, Varsha KD (1996). Induction of systemic
Awasthi and Kumar 2003; Gupta et al., 2004). There are
resistance in plants against viruses by a basic protein from
also reports of in vitro induction of these phytoprotein and
Clerodendrum aculeatum leaves. Phytopathology 86: 485-492.
Verma HN, Awasthi LP, Mukherjee K (1979) Induction of systemic
enhancement of growth in certain selected plants (Verma
resistance by antiviral plant extracts in nonhypersensitive plants, J.
et al., 1979; Verma et al., 1984; Verma et al., 1996).
Plant Dis. Prot. 87: 735-740.
Mechanisms underlying this can be due the synthesis of
Verma HN, Choudhary B, Rastogi P (1984). Antiviral activity of leaf
a new proteinaceous virus inhibitory agent produced in
extracts of different Clerodendrum species. Z. Pflanzenkr
pflanzenschutz 91: 34-41.
cel s perceiving signals from systemic resistance
inducers. Apart from resistance induction, ribosome-
inactivating function of these antiviral proteins have also