AGROBACTERIUM MEDIATED GENETIC TRANSFORMATION IN RICE USING Cry 1Ac & Cry 1B genes
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Date
2004
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ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY RAJENDRANAGAR, HYDERABAD
Abstract
In the present investigation studies were carried out to determine the callus
induction, growth and regeneration in two indica rice genotypes viz. Swarna and
APMS6B. Studies were conducted to optimize various parameters for stable
transformation in rice through Agrobacterium mediated gene transfer.
Callus induction frequency in MS medium in light was higher than in dark
irrespective of the genotype. Regeneration in both Swarna and APMS6B did not show
much variation between calli induced in MS and N6 basal medium and incubation in
light and dark.
Transformation of calli derived from mature seeds of two genotypes was
attempted using Agrobacterium EHA105/1304 plasmid containing Cry1AC and gus
reporter gene was used for transformation studies. Gus expression was observed in
two genotypes. Success of plant transformation using Agrobacterium depends on
factors such as (i) concentration of Acetosyringone (ii) Concentration of
Agrobacterium cells in culture. (iii) infiltration time and (iv) hygromycin in selection
medium. Concentration of Acetosyringone, 100 M and concentration of
Agrobacterium cells, 2 x 108 cells/ml (OD = 2) with infiltration time of 30 minutes
showed higher transformation rate. Present study thus indicates that genetic
transformation in rice can be successfully achieved by optimizing various parameters
for Agrobacterium mediated transformation. In the molecular detection study, F2
segregants of cross between BPT 211 and IR72 with a Xa21 were analysed for the
native Xa21 gene by homozygous and heterozygous condition.
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Keywords
AGROBACTERIUM, MEDIATED, GENETIC, TRANSFORMATION, RICE, Cry, 1Ac & Cry 1B genes,