Somatic Embryogenesis and Agrobacterium-mediated Transformation In Chickpea (Cicer arietinum L.)
Kiran S. Ghanti
Plant Tissue Culture Laboratory, Department of Botany Gulbarga University, Gulbarga-585 106, India.
Srinath Rao
Plant Tissue Culture Laboratory, Department of Botany Gulbarga University, Gulbarga-585 106, India.
R. B. Venugopal
Plant Tissue Culture Laboratory, Department of Botany Gulbarga University, Gulbarga-585 106, India.
S. Ramgopal Rao
Plant Tissue Culture Laboratory, Department of Botany Gulbarga University, Gulbarga-585 106, India.
C. P. Kaviraj
Plant Tissue Culture Laboratory, Department of Botany Gulbarga University, Gulbarga-585 106, India.
Arvind Barad
Plant Tissue Culture Laboratory, Department of Botany Gulbarga University, Gulbarga-585 106, India.
*Author to whom correspondence should be addressed.
Abstract
Agrobacterium-mediated transformation of Cicer arietinum L. was achieved from ICCV-10 and Annigeri varieties via somatic embryogenesis using mature and immature cotyledons, on MS medium fortified with B5 vitamins, different concentration of 2,4-D, 2,4,5-T and NAA in combination with BAP. The well formed embryos germinations of BAP and Zeatin (0.5-2.0 mg/l). Regenerated plantlets were transferred to pots, but they did not survive. Agrobacterium tumefaciens strain LBA 4404 carrying a binary vector containing NPT-II gene as selectable marker and GUS as a reporter gene was used for transformation. Histochemical analysis for GUS gene expression was observed as a primary evidence for transformation.
Keywords: Chickpea, Somatic embryogenesis, GUS, Kanamycin, NPT-II, Transformation