Novel Technique To Develop Transgenic Selectable Marker Free Pigeon Pea (Cajanus Cajan) Conferring Resistance Against Pod Borer Helicoverpa Armigera | 60092
Journal of Biotechnology & Biomaterials
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Pigeon pea is one of the major grain legumes of tropics and subtropics, covering vast regions of developing countries from Africa,
Asia to Latin America. It ranks fifth in area among pulses after soybean, common bean, peanut and chickpea. Globally, pigeon pea
is cultivated on 4.92 million hectares with an annual production of 3.65 metric tons and productivity of 898 kg/ha2. As they are grown
in harsh environments and exposed to a variety of biotic and abiotic stresses, their productivity has not increased conspicuously
for the last 50 years. Among many insect pests, the pod borer Helicoverpa armigera causes significant damage to this crop. It is the
most devastating Lepidopteran pest and causes extensive economic losses to the tune of US$ 300 million annually. The present study
seeks to protect pigeon pea plants from H. armigera infestation by incorporating cry1Ac and cry2Aa genes, through a unique and
efficient gene transfer method. An Agrobacterium tumefaciens-mediated transformation strategy was implemented using in vitro
transgenic shoot-grafting technique. A. tumefaciens harboring different binary vectors containing cry1Ac and cry2Aa genes were
used for transgenic pigeon pea development. An overall 7-9% of transformation frequency was recorded. After monitoring transgene
integration by Southern hybridization, transgenic T1 and T2 lines were further analyzed by western blot, ELISA and insect bioassay.
Transgenic lines obtained, exhibited optimum expression of Cry1Ac and Cry2Aa proteins. This study was further extended to the
development of selectable marker (nptII) free cry1Ac expressing transgenic lines using cre-lox mediated marker elimination system.
Gourab Ghosh is currently pursuing his PhD in Transgenic Crop Science from St. Xavier’s College, University of Calcutta, India. He has two publications in reputed, peer-review journals to his credit.