Indian Society of Genetics & Plant Breeding
In the current investigation experiments were performed to
produce high yielding transgenic rice plants, suitable for
upland paddy cultivation in Gujarat. DREB 2A gene isolated
from drought tolerant rice variety was placed in a binary
vector driven by 35S promoter of cauliflower mosaic virus
(CaMV) and a 5’non-coding region (5’-UTR) of rice alcohol
dehydrogenase (ADH) gene acting as translational
enhancer. Three to four weeks old proliferating calli of GR
11 variety of rice were used for transformation with
Agrobacterium strains LB 4404 harbouring pRION_DREB
2A plasmid. NPT II selectable marker gene in putatively
transformed plants confers resistance to kanamycin which
is used for selection of positively transformed GR 11 callus.
In vitro regeneration of the transformed callus produced T0
lines of rice harbouring DREB 2A cassette. The frequency
of transformation was 10%. Stable integration of transgene
conferring tolerance to drought in T0 rice plants were verified
using PCR, Reverse transcription PCR (RT PCR), Real Time
PCR (absolute and relative quantification). The transgenic
copy number in T0 cisgenic lines were calculated as one to
three copies of stably integrated DREB 2A gene per copy of
genome of GR 11 rice variety.
Keywords: In vitro regeneration, indica rice, Agrobacterium tumefaciens, genetic transformation, cisgenic plants
Year: 2015
Volume: 75
Issue: 1
Article DOI: 10.5958/0975-6906.2015.00003.6
Print ISSN: 0019-5200
Online ISSN: 0975-6906
R. Sandeep Raj info_circle
Chandrakant Singh info_circle
Arpan Modi and N. Subhash info_circle
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