<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated genetic transformation of pigeon pea [<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp.] for resistance to legume pod borer <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> |
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| Authors: | Email author" target="_blank">Gaurav?KrishnaEmail author P?Sairam?Reddy Pramod?W?Ramteke Pogiri?Rambabu Kailas?Bhagawanrao?Tawar Parthasarathi?Bhattacharya |
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| Institution: | 1.College of Biotechnology & Allied Sciences,Allahabad Agricultural Institute Deemed University,Allahabad,UP, India;2.Biotechnology Division,JK Agri Genetics Ltd.,Begumpet,AP, India |
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| Abstract: | Agrobacterium-mediated genetic transformation was performed using embryonic axes explants of pigeon pea. Both legume pod borer resistant
gene (cry1Ac) and plant selectable marker neomycine phosphor transferase (nptII) genes under the constitutive expression of the cauliflower mosaic virus 35S promoter (CaMV35S) assembled in pPZP211 binary
vector were used for the experiments. An optimum average of 44.61% successfully hardened dot blot Southern hybridization positive
plants were obtained on co-cultivation media supplemented with 200 μM acetosyringone without L-cysteine. The increased transformation
efficiency from a baseline of 11.53% without acetosyringone to 44.61% with acetosyringone was further declined with the addition
of different concentrations of L-cysteine to co-cultivation media. Transgenic shoots were selected on 50 and 75 mg L−1 kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 20 g L−1 sucrose and 0.5 mg L−1 indole butyric acid in the absence of kanamycin. Furthermore, 100% seed setting was found among all the transgenic events.
The plants obtained were subjected to multi- and nochoice tests to determine the behavioral responses and mortality through
Helicoverpa armigera bioassays on the leaf and relate their relationship with the expression of cry1Ac protein which was found to be less in leaf as compared to the floral buds, anther, pod, and seed. |
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