Agrobacterium-mediated transformation of indica rice cultivars using binary and superbinary vectors

Khanna, Harjeet K. & Raina, Satish K. (1999) Agrobacterium-mediated transformation of indica rice cultivars using binary and superbinary vectors. Australian Journal of Plant Physiology, 26, pp. 311-324.

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The Agrobacterium-mediated transformation system was extended to two indica cultivars: a widely cultivated breeding line IR-64 and an elite basmati cultivar Karnal Local. Root tips and shoot tips of seedlings, and scutellar-calli derived from mature seeds showed high-efficiency Agrobacterium tumefaciens infection and stable transformation. In addition to the superbinary vector pTOK233 in Agrobacterium strain LBA4404, almost equally high levels of transformation were achieved with a relatively much smaller (13.1 kb) binary vector (pCAMBIA1301) in a supervirulent host strain AGL1. In both cases, as well as in both cultivars, while 60–90% of the infected explants produced calli resistant to the selectable agent hygromycin, 59–75% of such calli tested positive for GUS. A high level (400 μM) of acetosyringone in the preinduction medium for Agrobacterium and a higher level (500 μM) in the cocultivation medium was necessary for an enhancement in transformation frequency of the binary vector to levels comparable to a superbinary. Hygromycin-resistant calli could be produced from all the explants used. Transformants could be regenerated for both cultivars using the superbinary and binary vector, but only for calli of scutellar origin. In addition to the molecular confirmation of hpt and gus gene transfer and transcription, absence of gene sequences outside the transferred DNA (T-DNA) region confirmed absence of any long T-DNA transfer.

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ID Code: 50736
Item Type: Journal Article
Refereed: Yes
Keywords: Agrobacterium-mediated transformation, indica rice, superbinary vector, cocultivation, basmati
DOI: 10.1071/PP98160
ISSN: 1445-4408
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000)
Divisions: Current > Research Centres > Centre for Tropical Crops and Biocommodities
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Copyright Owner: Copyright 1999 CSIRO Publishing
Deposited On: 05 Jun 2012 01:13
Last Modified: 05 Jun 2012 01:13

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