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20071
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Subject: Biotechnology and Molecular Biology × Author: Chowdhary, Ritika × End date: 2009 × Start date: 2004 ×
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    ThesisItemOpen Access
    Agrobacterium mediated transformation of indica rice variety IR-72 using Amaranthus seed albumin gene (AmA1) driven by a seed specific promoter
    (CCSHAU, 2007) Chowdhary, Ritika; Jain, R.K.
    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.