Biologia plantarum 51:61-68, 2007 | DOI: 10.1007/s10535-007-0012-1

Agrobacterium-mediated high frequency transformation in dwarf recalcitrant rice cultivars

P. Kant1,*, S. Kant1, R. K. Jain2, V. K. Chaudhury2
1 J. Blaustein Institutes for Desert Research, Department of Dryland Biotechnologies, Ben-Gurion University of the Negev, Midreshet Ben-Gurion, Israel
2 Dept. of Biotechnology and Molecular Biology, College of Basic Sciences and Humanities, CCS Haryana Agricultural University, Hisar, India

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.

Keywords: Bacillus thuringiensis endotoxin; hygromycin; kinetin; napthaleneacetic acid; PCR; potato proteinase inhibitor
Subjects: abscisic acid (ABA); Agrobacterium tumefaciens; auxins; Bacillus thuringiensis endotoxin; cultivar and genotype differences; cytokinins; in vitro culture, callus growth and duration; in vitro culture, regeneration, proliferation, differentiation; nutrient medium, Murashige and Skoog (MS); Oryza sativa; polymerase chain reaction (PCR); protease; recalcitrant dwarf cultivars; transgenic plants

Received: March 4, 2005; Accepted: January 15, 2006; Published: March 1, 2007Show citation

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Kant, P., Kant, S., Jain, R.K., & Chaudhury, V.K. (2007). Agrobacterium-mediated high frequency transformation in dwarf recalcitrant rice cultivars. Biologia plantarum, 51(1), 61-68. doi: 10.1007/s10535-007-0012-1.
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