Agrobacterium-Mediated Genetic Transformation in Indian Spinach (Beta palonga)
Gaurab Gangopadhyay
Cytogenetics Laboratory, Department of Botany Bose Institute, 93/1 A.P.C. Road, Calcutta 700 009, West Bengal, India
Tanoy Bandyopadhyay
Cytogenetics Laboratory, Department of Botany Bose Institute, 93/1 A.P.C. Road, Calcutta 700 009, West Bengal, India
Siraj Datta
Cytogenetics Laboratory, Department of Botany Bose Institute, 93/1 A.P.C. Road, Calcutta 700 009, West Bengal, India
Debabrata Basu
Cytogenetics Laboratory, Department of Botany Bose Institute, 93/1 A.P.C. Road, Calcutta 700 009, West Bengal, India
Kalyan K Mukherjee *
Cytogenetics Laboratory, Department of Botany Bose Institute, 93/1 A.P.C. Road, Calcutta 700 009, West Bengal, India
*Author to whom correspondence should be addressed.
Abstract
An efficient and reproducible protocol for Agrobacterium-mediated plant transformation has been standardized in Indian spinach (Beta palonga) with a standard vector (pCambia 1304). Passage wise subculture in medium containing hygromycin, the selection agent, eventually resulted in survival of two percent (eleven in number) of originally infected explants showing continuous organogenesis. Of those eleven lines only three showed GUS amplification after PCR analysis followed by GUS positive histochemical assay. Those three confirmed transgenics were multiplied followed by successful rooting in liquid medium with coir as alternative matrix.
Keywords: Beta palonga, Genetic transformation, GUS amplification, Hygromycin resistance