We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
An improved Agrobacterium-mediated transformation of recalcitrant indica rice ( Oryza sativa L.) cultivars.
- Authors
Shri, Manju; Rai, Arti; Verma, Pankaj; Misra, Prashant; Dubey, Sonali; Kumar, Smita; Verma, Sikha; Gautam, Neelam; Tripathi, Rudra; Trivedi, Prabodh; Chakrabarty, Debasis
- Abstract
Agrobacterium-mediated transformation of indica rice varieties has been quite difficult as these are recalcitrant to in vitro responses. In the present study, we established a high-efficiency Agrobacterium tumefaciens-mediated transformation system of rice ( Oryza sativa L. ssp. indica) cv. IR-64, Lalat, and IET-4786. Agrobacterium strain EHA-101 harboring binary vector pIG121-Hm, containing a gene encoding for β-glucuronidase (GUS) and hygromycin resistance, was used in the transformation experiments. Manipulation of different concentrations of acetosyringone, days of co-culture period, bacterial suspension of different optical densities (ODs), and the concentrations of l-cysteine in liquid followed by solid co-culture medium was done for establishing the protocol. Among the different co-culture periods, 5 days of co-culture with bacterial cells (OD = 0.5-0.8) promoted the highest frequency of transformation (83.04 %) in medium containing l-cysteine (400 mg l). Putative transformed plants were analyzed for the presence of a transgene through genomic PCR and GUS histochemical analyses. Our results also suggest that different cultural conditions and the addition of l-cysteine in the co-culture medium improve the Agrobacterium-mediated transformation frequencies from an average of 12.82 % to 33.33 % in different indica rice cultivars.
- Subjects
RICE varieties; ORYZA; AGROBACTERIUM; AGROBACTERIUM tumefaciens; CULTIVARS
- Publication
Protoplasma, 2013, Vol 250, Issue 2, p631
- ISSN
0033-183X
- Publication type
Article
- DOI
10.1007/s00709-012-0439-x