Using Waxy locus to reveal mutation in TRIM rice population
Date Issued
2008
Date
2008
Author(s)
Chen, Yi-Fang
Abstract
Rice（Oryza sativa L.）is one of the most important crops in the world. T-DNA has been used successfully in rice functional genomics studies. For rice transformations, the integration of T-DNA occurred at the callus tissue, as a consequence, somaclonal variations might be generated during cultured period. The tagging efficiency of T-DNA induced population is relatively low, that is, about 5-10％ of the phenotype co-segregate with the T-DNA integration site. In the present research, TILLING, targeting induced local lesion in genomes, a novel technique with high throughput discovery of single nucleotide changes, was used to detect SNPs, single nucleotide polymorphism, in Waxy locus between two japonica varities, Tainung 67 and Nipponbare. In Taiwan, Hsing et al.,（2007）used T-DNA transform to build up a mutant population using an elite local variety Tainung 67 and designated TRIM, Taiwan Rice Insertion Mutants. In the present research, TILLING was used to reveal Waxy locus muataions in TRIM population, in order to characterize the frequency and type of somacloal variations. o SNPs are present in Waxy locus（6297 bp） between Tainung 67 and Nipponbare cultivars. For 200 TRIM lines, or 3000 individual plants, five lines consisted somaclonal variations in Waxy locus. One of these lines contained two SNPs, thus, totally 6 SNPs were detected. All SNPs were belong to nucleotide substitution, and they located in the promoter or 5，-UTR region in Waxy gene. To conclude, 6 nucleotide substitutions was found , after screening 3000 plants × 6297 bp Waxy gene. The mutation frequency is estimated as 3.1×10-7.
Subjects
somacloal variation
Type
thesis
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