Agrobacterium-mediated transfer of amaranthus seed albumin (AmA1) gene driven by seed specific promoter in indica rice variety (HKR-46) for improving the protein quality
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Date
2007
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Publisher
CCSHAU
Abstract
Experiments were conducted to transfer Amaranthus seed
albumin gene, AmA1, driven by GBSS promoter in indica rice variety HKR46
by Agrobacterium-mediated transformation. The japonica rice variety TNG67
was also used as control for transformation. During the present investigation,
21-24 days old calli induced from mature seed scutella and immature
embryos on modified MS medium were used. The calli were co-cultivated for
10 minutes in liquid co-cultivation medium with Agrobacterium strain
EHA105 (pSB8G) and then transferred onto the solidified co-cultivation
medium for 2-3 days. Then the calli were screened on selection medium
containing cefotaxime (250 mg/l) and G-418 (50 mg/l) for 2-3 cycles of
selection of 15 days each. A total of 23/1822 calli of HKR46 and 242/345 of
TNG67 showed sustained proliferation on the selection medium. The
putatively transformed calli were then transferred onto regeneration medium.
A total of 9 HKR-46 and 89 TNG-67 plants were regenerated from the selected
calli by using the MS based regeneration media supplemented with
antibiotics. Eighteen of these plants (HKR-46, 4 plants; TNG-67, 14 plants)
were tested for desired bands. An amplified product of 1.1 kb fragment was
detected in 13 plants (Table 4.4).
The study shows an urgent need of developing an efficient protocol for high
frequency plant regeneration of transformed indica and japonica rice
varieties.
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Keywords
Rice, Transformation, AmA1 gene, Seed specific promoter