Today, there are numerous reports and articles written about the transformation of recalcitrant Indica rice cultivars such as IR17 and IR64. While most of these reports suggest that these recalcitrant indica rice cultivars have a very low frequency of transformation, the hypothesis of this experiment is to suggest that the transformation protocol can easily be improved when mature seed-derived calli are used as explants. The plant materials used in this experiment were mature seeds of rice cultivars – IR72 and IR64.
The experiment showed that the most essential factor towards increasing efficiency of transformation is the starting material for the process. While earlier researchers used calli that were 3 weeks old, this experiment uses 2 month old embryogenic calli. The experiment also sensitizes the formation of embryogenic calli (Kumar, Maruthasalam, Loganathan, Sudhakar & Balasubramanian, 2005) . In addition to that, there is also an emphasize on use of maltose as the source of carbon, the use of a gelling agent and 2,4-D that are of a high percentage. As the graph illustrates, a transformation efficiency of between 4.6% and 5.5% was obtained in IR64. The graph also illustrates that for IR72, a transformation efficiency between 6.4% and 7.3% was obtained.
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For an effective transformation of recalcitrant indica rice genotype, the experiment found that there is a need to utilize scutellar derived embryogenic calli that is about 2 months old. The experiments also note that the embryogenic calli should be used on a medium that has a higher concentration of the gelling agent. A high concentration of gelling agent is said to increase greatly increase the quality of 2,4-D. in addition to that, the high concentration also increases the quality of maltose not only as a source of carbon but also in helping reduce injuries to the explants while in the process of cocultivation. For the next experiment, researchers could consider extending the protocol of the current experiment for the transformation of any other recalcitrant indica rice cultivars apart from IR72 and IR64.
Reference
Kumar, K., Maruthasalam, S., Loganathan, M., Sudhakar, D., & Balasubramanian, P. (2005). An improvedAgrobacterium-mediated transformation protocol for recalcitrant elite indica rice cultivars. Plant Molecular Biology Reporter , 23 (1), 67-73. doi: 10.1007/bf02772648
