Agrobacterium mediated transformation of indica rice variety HKR46 with amaranthus seed albumin gene, AmA1
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Date
2006
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CCSHAU
Abstract
Experiments were conducted to transfer the Amarantus seed
albumin gene, AmA1, driven by CaMV35S 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, 15-18 days old calli initiated 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 (pSB8) and then
transferred onto the solidified co-cultivation medium for two days. After
two days the co-cultivated calli were transferred onto selection medium
containing kanamycin/paromomycin (50 mg/l) for 2-3 cycles of
selection of 15 days each. A total of 13/1717 and 103/278 calli of
HKR46 and TNG67 respectively showed sustained proliferation in the
selection medium. The putatively transformed calli were then
transferred onto regeneration medium. No shoot regeneration was
obtained in case of indica rice variety HKR46. However, eight of the
putatively transformed calli of TNG67 regenerated shoots; but all of
these shoots were albino. The study shows an urgent need of
developing an efficient protocol for high frequency green plant
regeneration/genetic transformation in indica rice varieties.