Micropropagation and anther/pollen culture of Jatropha curcas: A source for biodiesel
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Date
2008
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Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
Jatropha curcas L. is oil yielding tree plant with high medicinal and commercial
value. This plant has been identified as the promising plant for biodiesel production by
Planning Commision’s Task Force. As Jatropha is cross-pollinated crop so, genetic
integrity of the plant can’t be maintained through seed germination. The in vitro culture
is one of the best techniques for mass propagation and crop improvement to increase
productivity leading to full supply of the demand. In the present work, an efficient in
vitro method for plantlet regeneration via shoot tips and leaves of J. curcas (variety-
Mancheswar) was developed. The best survival percentage during surface sterilization of
J. curcas (variety- Mancheswar) explants were achieved by treating shoot tips with 0.1%
of HgCl2 for 10 minutes and leaves with 0.05% HgCl2 for 10 minutes. Shoot
multiplication was induced on MS media supplemented with 2.0 mg/l of Kn, 1.5 mg/l
IBA, 25.0 mg/l AdSO4, 10.0 mg/l ascorcic acid and 50.0 mg/l of citric acid. Highest
number of shoots per explant (5-7 shoots) was observed after 60 days of inculation in the
same media. Best callus growth was observed on MS media containing 1.0 mg/l of 2,4-
D, 1.0 mg/l of kinetin and 1.0 mg/l of citric acid. Best shoot regeneration from callus was
observed on MS media containing BAP (1.5 mg/l), kinetin (0.5 mg/l) and IAA (0.125
mg/l) as well as MS medium supplemented with 1.0 mg/l of BAP and 3.0 mg/l of 2-ip
after 60 days of inoculation (4-6 shoots per callus clump). No root formation was
observed under in vitro system. For rooting, a pulse treatment of excised shootlets, 10.0
mg/l of IBA was found effective under in vivo condition.
In vitro anther/pollen culture is an important technique to induce haploids either
by direct embryo formation or through callus formation. So, efforts have been made to
establish culture by anther/pollen of J. curcas (variety- Kalyanpur and PKVJ MKVI).
Callus was successfully induced from anther of both varieties of J. curcas on MS media
supplemented with 1.0 mg/l of 2,4-D, 1.0 mg/l of kinetin and 1.0 mg/l of CA. No shoot
regeneration from anther was observed in MS media supplemented with different
hormonal concentrations and combinations. Only white globular structures were
observed all over the callus clumps after 20 days of transfer on MS media supplemented
with 1.0 mg/l of 2,4-D, 1.0 mg/l of kinetin, but shoot regeneration was not observed.
Protoplasts were successfully isolated from both varieties of callus of J. curcas (variety-
Kalyanpur and PKVJ MKVI) but protoplasts culture could not be established in liquid as
well as solid NT and B5 media.
In living organisms the reactive oxygen species (ROS) are known to cause
damage to lipids, proteins, enzymes and nucleic acids leading to cell or tissue injury,
which have been implicated in the processes of aging as well as in wide range of
degenerative diseases. Many Indian medicinal plants are potential source of antioxidants.
Natural antioxidants are secondary metabolites of plants that neutralize these oxidants. In
the present work, phytochemical tests were done for the identification of different
chemical constituents of J. curcas (var.- Mancheswar). The presence of different
chemical constituents such as alkaloids, sugars, terpenoids, steroids, aminoacids,
phytosterols, saponins, tannins and phenols from J. curcas was confirmed by
phytochemical tests. Screening of antioxidant activity of the crude methanolic extracts of
J. curcas (var.- Mancheswar) was done by using the spectrophotometric DPPH (1,1-
diphenyl-2-picrylhydrazyl) as well as ABTS (2,2’-azinobis-(3-ethylbenzthiazoline-6-
sulphonic acid) free radical scavenging method and DNA damage protecting activity was
checked by inducing hydroxyl radicals on pBS SK II (-). Antioxidant activity by DPPH
and ABTS assay showed that 50 μg/ml of crude methanolic extract of J. curcas was able
to inhibit the DPPH radical formation by 91.92% and 30 μg/ml of plant extract was able
to inhibit ABTS radical formation by 98.84% respectively. Addition of 20 μg/μl of plant
extract to the reaction mixture of H2O2 indicated the significant protection to the damage
of pBS SK II (-).
Description
Micropropagation and anther/pollen culture of Jatropha curcas: A source for biodiesel
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