Journal of
Plant Breeding and Crop Science

  • Abbreviation: J. Plant Breed. Crop Sci.
  • Language: English
  • ISSN: 2006-9758
  • DOI: 10.5897/JPBCS
  • Start Year: 2009
  • Published Articles: 409

Full Length Research Paper

Transformation of chickpea lines with Cry1X using in planta transformation and characterization of putative transformants T1 lines for molecular and biochemical characters

Asharani, B. M.*, Ganeshaiah, K. N., Amrita Raja Vinod Kumar and Udayakumar Makarla
Department of Genetics and Plant Breeding, University of Agricultural Sciences, Bangalore-560 065, India.
Email: [email protected]

  •  Published: 30 December 2011

Abstract

 

The transgenic plants of the chickpea (cicer arietinum) cv. KAK-2 expressing a Bt gene, were generated using an Agrobacterium tumefaciens mediated transformation system. A tissue culture-independent transformation method, in planta which targets the A. tumefaciens to the apical meristem was used in this study. The protocol involves in planta inoculation of the embryo axes of the germinating seeds and allowing them to grow into seedlings ex vitroPolymerase chain reaction (PCR) analysis indicated the putative transgenic nature of theT1 generation plants. Bioassays against major pests of the chickpea, Helicoverpa armigera revealed several T1 plants that perform well against the larvae. This revealed that 43 of T1 plants harbor the transgene. The seeds of 43 T1 plants were allowed to continue into the next generation amplified gene of interest in most of the plants tested. Enzyme Linked-Immuno Sorbent Assay (ELISA), PCR and Bioassays was used to identify the high expressing plants. These results suggest that the Bt gene was functional in the transgenic chickpea and was being expressed. The study also showed that the chickpea plants harboring the cry1X gene were resistant toHelicoverpa armigera.

 

Key words: Chickpea, Helicoverpa armigera, transformation, in-planta, tissue culture, synthetic cry gene.

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