EFFICIENT TISSUE CULTURE INDEPENDENT Agrobacterium MEDIATED GENETIC TRANSFORMATION PROTOCOLS FOR Solanum lycopersicum
ATUL GROVER *
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India
SADHANA SINGH
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India
PANKAJ PANDEY
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India and National Agri-Food Biotechnology Institute, C-127, Industrial Area, S.A.S. Nagar, Phase 8, Mohali, Punjab 160071, India
PATADE VIKAS YADAV
Defence Institute of Bio-Energy Research Field Station Pithoragarh, Panda Farm, Pithoragarh, Uttarakhand 263145, India
SANJAY MOHAN GUPTA
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India
ANJILI YADAV
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India and Department of Bioscience and Biotechnology, Banasthali University, Banasthali, Rajasthan 304022, India
AAKANKSHA JADON
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India and Department of Bioscience and Biotechnology, Banasthali University, Banasthali, Rajasthan 304022, India
HIMANSHU BHATT
Department of Biotechnology, Motiram Baburam Government Post Graduate College, Haldwani, Uttarakhand 263139, India
OM PRAKASH
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India
BASANT BALLABH BHATT
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India
MOHAMMED NASIM
Defence Institute of Bio-Energy Research, Goraparao, P.O. Arjunpur, Haldwani, Uttarakhand 263139, India
*Author to whom correspondence should be addressed.
Abstract
Aims: Efficient and rapid in planta genetic transformation methods using Agrobacterium tumefaciens for tomato have been proposed.
Place and Duration of Study: Defence Institute of Bio Energy Research, Haldwani, India between June and December 2013.
Methodology: In one of the methods, Agrobacterium tumefaciens culture containing recombinant plasmids, suspended in MS medium containing 100 µM Acetosyringone, was injected in locules of ripened fruits. In another method, tomato seedlings were transferred to YEM medium containing 100 µM Acetosyringone, 10 mg/l Rifampicin along with Agrobacterium tumefaciens culture harbouring recombinant plasmids. The culture was incubated overnight at 27°C and 200 rpm.
Results: Using each of these methods, transformation efficiencies in the range 49-73% were obtained in the progenitor generation.
Conclusion: Direct contact of Agrobacterium with developing embryo or with germinating seed has been found easy and efficient approach to get transformants.
Keywords: Tomato, in planta transformation, Agrobacterium tumefaciens, real-time PCR