Agrobacterium mediated transformation of indica rice variety IR-72 using Amaranthus seed albumin gene (AmA1) driven by a seed specific promoter
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Date
2007
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CCSHAU
Abstract
Experiments were conducted to transfer Amaranthus seed
albumin gene (AmA1) driven by GBSS promoter in indica rice variety IR-72
and Pusa Basmati by Agrobacterium-mediated transformation. During the
present investigation, 14-18 days old calli induced from mature seed scutella
and immature embryos on modified MS medium were used. The calli were
II
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. 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 160/1790 (mature seed) and 37/213
(immature embryo) calli of IR-72 and 124/831 of Pusa Basmati 1 showed
sustained proliferation on the selection medium. A total of 22 IR72 and three
Pusa Basmati 1 plants were regenerated from the selected calli by using the
MS based regeneration media supplemented with antibiotics. Fifteen of these
plants (IR72, 12 plants; Pusa Basmati 1, three plants) were successfully
transferred to the transgenic greenhouse. Five of the seven-rice plants
showed the amplification of 1.1 kb fragment indicating the presence of AmA1
gene. One of these PCR positive plants has already flowered and set seeds.
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Keywords
Rice, Transformation, AmA1 gene, seed specific promoter