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GrainGenes Reference Report: TAG-107-84

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Reference	TAG-107-84
Title	Detection of single nucleotide polymorphism (SNP) controlling the waxy character in wheat by using a derived cleaved amplified polymorphic sequence (dCAPS) marker
Journal	Theoretical and Applied Genetics
Year	2003
Volume	107
Pages	84-88
Author	Yanagisawa T Kiribuchi-Otobe C Hirano H Suzuki Y Fujita M
Abstract	Summary: We investigated a single nucleotide polymorphism (SNP) in the Wx-D1 gene, which was found in a mutant waxy wheat, and which expressed the Wx-D1 protein (granule-bound starch synthase I) as shown by immunoblot analysis. We also assayed starch synthase activity of granule-bound proteins. Using 22 doubled-haploid (DH) lines and 172 F5 lines derived from the wild type x the mutant, we detected SNP via a PCR-based (dCAPS) marker. Amplified PCR products from Wx-D1 gene-specific primers, followed by mismatched primers designed for dCAPS analysis, were digested with the appropriate restriction enzyme. The two alleles, and the heterozygote genotype were easily and rapidly discriminated by gel-electrophoresis resolution to reveal SNP. All progeny lines that have the SNP of the mutant allele were waxy. Integrating the results of dCAPS analysis, immunoblot analysis and assays of starch synthase activity of granule-bound proteins indicates that the SNP in the Wx-D1 gene was responsible for its waxy character. This dCAPS marker is therefore useful as a marker to introduce the mutant allele into elite breeding lines
External Databases	Pubmed: 12669198
Keyword	[ Hide all but 1 of 37 ] allele assay bound starch synthase breeding dcaps marker detection doubled haploid element endosperm enzyme gene genetic analysis genotype granule-bound starch synthase locus maize marker mutant pcr polymorphism primer products progeny restriction sequence single nucleotide polymorphism snp starch starch synthase starches synthase synthase-i waxy waxy gene waxy wheat wild wild-type

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