Silencing of a rice germin-like protein (GLP) increases the drought tolerance in indica rice cultivar
2013
Biswal, A.K. | Min, A. | Tsakirpaloglou, N. | Slamet-Loedin, I.H. | Kohli, A.
Germin-like protein (GLP)s are ubiquitous plant proteins largely associated with abiotic/biotic stress response, plant growth and development. A 23k D rice GLP gene, located on chromosome 8 (LOC_Os08g08960), was earlier reported to be developmentally regulated. To further characterize the role of this gene in abiotic stress tolerance, the coding sequence of the OsGLP8960 gene was cloned and over expressed in indica rice cultivar IR64 through Agrobacterium-mediated genetic transformation. The target gene was also silenced through post-transcriptional gene silencing (PTGS) by transforming IR64 with an artificial miRNA (amiRNA) construct in similar manner. Two each of single copy insertion Tsubo transgenic lines which had similar morphology as the wild type were selected for further generation advancement. The Tsub1 transgenic lines showed 3:1 genetic segregation, expected for single copy and stable transgene integration. Drought stress was applied by withholding watering for two weeks in vegetative stage and for 10 days in reproductive stage. IR64 and Nipponbare were used as the susceptible check while Apo was used as tolerant check. Followed by Nipponbare, IR64 and the over expression lines showed high level of susceptibility while the silencing lines showed lower leaf rolling and faster recovery after stress as Apo. The leaf relative water content (RWC) of silencing lines was the highest followed by Apo while rest of the lines including the over expression line were almost dry with very low RWC. Preliminary drought screening at reproductive stage revealed similar leaf rolling pattern. Further physiological and agronomic characterization including spikelet fertility and grain weight measurements are in progress.
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