Double haploidy as a route to generating homozygous insertion mutants of a germin-like protein to understood its role in rice endosperm development
2013
Min, A. | Azanes, M.D. | Dane, R.H. | Tsakirpaloglou, N. | Slamet-Leodin, I. | Kohli, A.
Ubiquitous family of germin and germin-like plant proteins are known to function as superoxide dismutase (SOD). In rice, a 23kD rice GLP gene which is located on chromosome 8 (LOC_Os08g08960) was earlier reported to be highly upregulated in the spikelet. A single copy transposon insertion mutant in Osglp8960 was obtained. The expected 3:1 segregation of the insertion was not observed. Instead 1:2 ratio of normal to heterozygous mutant was obtained. This indicated homozygous mutation lethality. To underpin the role of Osglp8960, the heterozygous mutant plants were used in pollen culture to induce double haploidy mediated homozygous insertion mutant. Homozygous mutant lines were then screened for germination, seedling vigor and grain filling ability. Results showed that homozygous mutants had significantly lower germination rate than the wild type Nipponbare. Seedling vigor test in 10 day old seedlings showed that homozygous lines had shorter root and shoot length. At harvest, it was also found that the spikelet fertility of mutant lined decreased by two-folds. These data suggest that GLP Os08g08960 plays an important role in germination, seedling vigor and grain filling.
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