Genetic Evaluation of Agrobacterium rhizogenes mediated Hairy Root induction on Phlogacanthus thyrsiflorus Nees
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Date
2011
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Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
The natural tendency of Agrobacterium rhizogenes, to introduce a part of its DNA
called T-DNA into the plant cells resulting into production of neo plastic roots
called hairy roots, make them a plausible choice for their use in plant
biotechnological applications. Their fast growth, genetic and biosynthetic stability,
ease of maintenance and ability to synthesize a range of chemical compounds
that makes them a suitable system for in vitro production of secondary
metabolites. Phlogacanthus thyrsiflorus Nees, a valued but comparatively
unexplored medicinal plant has been tested to explore its susceptibility to A.
rhizogenes for acceleration of useful secondary metabolites. They are genetically
evaluated to confirm the complete and stable genetic transformation.
PCR analysis of DNA from hairy roots induced on Phlogcanthus
thyrsiflorus with primer pairs for the amplification of, rolC, and aux1 genes were
performed. rolC and aux1 primer were used for carrying out PCR reaction.
Genomic DNA was extracted from transformed and non-transformed roots of P.
thyrsiflorus using standard CTAB method. Out of three distinctive annealing
temperatures i.e. 42°C, 46°C and 48°C employed for PCR, 46°C was found best
for band amplification. Amplification of aux1 and rolC primers at 815 and 487 bp
respectively was observed for the transformed root, whereas no amplification
was found for non-transformed roots. Thus the result revealed successful genetic
transformation of P. thyrsiflorus.
Description
Genetic Evaluation of Agrobacterium rhizogenes mediated Hairy Root induction on Phlogacanthus thyrsiflorus Nees
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