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Pigeon pea, a grain legume of the semiarid tropics, is a rich source of high-quality protein. The productivity of this pulse is seriously affected by lepidopteron insect pests. To generate a sustainable insect-resistant plant, synthetically prepared bioactive key constituents of a crystal protein (Syn Cry1Ab) of  Bacillus thuringiensis  were expressed in pigeon pea under the guidance of a tissue-specific promoter of the RuBP carboxylase/oxygenase small subunit ( rbcS ) gene. Regenerated transgenic plants with the  cry1Ab  expression cassette ( cry1Ab-lox-bar-lox ) showed the optimum insect motility rate (90%) in an in vitro insect bioassay with second instar larvae, signifying the insecticidal potency of Syn Cry1Ab. In parallel, another plant line was also generated with a chimaeric vector harbouring a  cre recombinase  gene under the control of the CaMV 2 × 35S promoter. Crossing between T 1  plants with a single insertion of  cry1Ab-lox-bar-lox  T-DNA and T 1  plants with moderate expression of a  cre  gene with a linked hygromycin resistance ( hptII ) gene was performed to exclude the bialaphos resistance ( bar ) marker gene. Excision of the  bar  gene was achieved in T 1 F 1  hybrids, with up to 35.71% recombination frequency. Insect-resistant pigeon pea plants devoid of selectable marker genes ( syn  Cry1Ab-  bar  and  cre-hptII ) were established in a consecutive generation (T 1 F 2 ) through genetic segregation.

Development of marker-free transgenic pigeon pea (Cajanus cajan) expressing a pod borer insecticidal protein