Development of STS markers and establishment of multiplex PCR for Glu-A3 alleles in common wheat (Triticum aestivum L.) |
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| Authors: | Linhai Wang Genying Li Roberto J Peña Xianchun Xia Zhonghu He |
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| Institution: | 1. Institute of Crop Science, National Wheat Improvement Centre/The National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences (CAAS), No. 12 Zhongguancun South Street, Beijing 100081, China;2. Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, Shandong, China;3. International Maize and Wheat Improvement Centre (CIMMYT), Apdo. Postal 6-641, 06600 Mexico, D.F., Mexico;4. International Maize and Wheat Improvement Centre (CIMMYT) China Office, c/o CAAS 12 Zhongguancun South Street, Beijing 100081, China |
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| Abstract: | Low-molecular-weight glutenin subunits (LMW-GS) play a key role in determining the processing quality of the end-use products of common wheat. The objectives of this study were to identify genes at Glu-A3 locus, develop the STS markers, and establish multiplex PCR with the STS markers for Glu-A3 alleles. Gene-specific PCR primers were designed to amplify six near-isogenic lines (NILs) and Glenlea with different Glu-A3 alleles (a, b, c, d, e, f and g) defined by the protein electrophoretic mobility. Three Glu-A3 genes with complete coding sequence were cloned, designated as GluA3-1, GluA3-2 and GluA3-3, respectively. Seven dominant allele-specific STS (sequence tagged sites) markers were designed based on the SNPs (single nucleotide polymorphisms) among different allelic variants for the discrimination of the Glu-A3 protein alleles a, b, c, d, e, f and g. Four multiplex PCRs were established including Glu-A3b + Glu-A3f, Glu-A3d + Glu-A3f, Glu-A3d + Glu-A3g, and Glu-A3b + Glu-A3e. These markers and multiplex-PCR systems were validated on 141 CIMMYT wheat varieties and advanced lines with different Glu-A3 alleles, confirming that they can be efficiently used in marker-assisted breeding. |
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| Keywords: | Common wheat LMW-GS Glu-A3 STS marker Multiplex PCR |
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