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Optimization of In-planta method of genetic transformation in pigeon pea (Cajanus cajan L. Millsp.)

Pigeon pea (Cajanus cajan L. Millsp.) is one of the major grain legume crops of the tropics and sub-tropics grown in about 50 countries of Asia, Eastern and Southern Africa and the Caribbean for various uses such as food, fodder and firewood. The present study was conducted to develop and optimize tissue culture independent in planta transformation system in pigeonpea. This system can help in widening genetic base as well as solutions over various biotic and abiotic factors through engineering novel genes across the species. The various transformation parameters viz., Optical density of Agrobacterium suspension, virulence inducer and infection time were optimizad through 18 different treatment combinations. The plumular and inter cotyledonary meristem axes of 2-3 days old germlings of pigeon pea cv. BSMR 853 was exploited for Agroinfection by sewing needles. Out of 270 inoculated germlings 11.34 mean number of plantlets were recovered. The putative transformants were confirmed by GUS histochemical and PCR assay. Among 18 different treatment combinations, the treatment pertaining Agrobacterium suspension of O.D 1.0, virulence inducer (acetosyringone) at 250 µM/ml and infection time of 1.0 min was found optimum has shown significant impact on transformation efficiency. The treatment comprising bacterial O.D, 1.0 with 250 µM/ml acetosyringone and 1.0 infection time 1.0 min revealed 90.02% transformation efficiency. However, lowest transformation frequency i.e. 68.76% was reported in treatment of bacterial O.D. 1.5 with 150 µM/ml acetosyringone and 0.5 min. infection time. The present investigation revealed the optimization of in planta transformation parameters in pigeon pea and suitability of genotype BSMR853 for genetic transformation and further genetic improvement. | Optimization of In-planta method of genetic transformation in pigeon pea (Cajanus cajan L. Millsp.)