Abstract
We report here a high-efficiency transformation protocol for recalcitrant indica rice cultivars IR64 and IR72 with the selectable marker genehph and thegusA reporter gene. Factors that favor high-efficiency transformation were found to be use of 2-month-old mature seed-derived embryogenic calli, maltose as a source of carbon, a higher concentration of 2,4-dichlorophenoxyacetic acid, and both phytagel and agar as gelling agents. The putative transgenic (T0) plants were analyzed for integration of the transgene through polymerase chain reaction and Southern blotting analyses. Various factors thought to be responsible for increased transformation efficiency are discussed.
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Abbreviations
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- BAP:
-
6-benzylaminopurine
- MS:
-
Murashige and Skoog
- NAA:
-
α-naphthaleneacetic acid
- OD:
-
optical density
- PCR:
-
polymerase chain reaction
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Kumar, K.K., Maruthasalam, S., Loganathan, M. et al. An improvedAgrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars. Plant Mol Biol Rep 23, 67–73 (2005). https://doi.org/10.1007/BF02772648
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DOI: https://doi.org/10.1007/BF02772648