Agrobacterium-mediated high frequency transformation in dwarf recalcitrant rice cultivars |
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Authors: | P. Kant S. Kant R. K. Jain V. K. Chaudhury |
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Affiliation: | (1) J. Blaustein Institutes for Desert Research, Department of Dryland Biotechnologies, Ben-Gurion University of the Negev, Midreshet Ben-Gurion, 84990, Israel;(2) Dept. of Biotechnology and Molecular Biology, College of Basic Sciences and Humanities, CCS Haryana Agricultural University, Hisar, 125004, India |
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Abstract: | The Agrobacterium-mediated transformation was done in rice (Oryza sativa L. var. indica) cv. HKR126 and elite cross-bred cv. Pusa Basmati1 (PB1), using strain LBA4404 containing pCAMBIA1300 cloned with gene cassettes; potato proteinase inhibitor and Bacillus thuringiensis endotoxin (plasmid JDW53) or mannitol-1-phosphate dehydrogenase (plasmid RKJ108). Co-cultivation with scutellar-calli derived from mature seeds showed stable and highly efficient transformation. In cvs. HKR126 and PB1, 35 % and 41 % of hygromycin resistant calli were obtained. The transformation efficiency in PB1 (22.0 %) was much higher than in HKR126 (12.5 %). Similarly, PB1 had higher plant regeneration efficiency than HKR126. The shoots regenerated per callus were, 3–4 in HKR126 and 5–6 in PB1. The transformation efficiency with pRKJ108 (18.6 %) was higher than pJDW53 (15.9 %). Polymerase chain reaction (PCR) analysis showed the presence of transgenes in regenerated transgenic plants of both cultivars. |
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Keywords: | Bacillus thuringiensis endotoxin hygromycin kinetin napthaleneacetic acid PCR potato proteinase inhibitor |
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