Development of simple and co-dominant PCR markers to genotype puroindoline a and b alleles for grain hardness in bread wheat (Triticum aestivum L.)

作者：Huang Xiu Qiang^*; Brule Babel Anita

来源：Journal of Cereal Science, 2011, 53(3): 277-284.

DOI：10.1016/j.jcs.2011.01.008

• Grain hardness
• Pina
• Pinb
• SNP (single nucleotide polymorphism)
• STS
• Temperature-switch (TS) PCR
• Triticum aestivum

Grain hardness is one of the most important quality characteristics of cultivated bread wheat (Triticum aestivum L). A large deletion in the puroindoline a (Pina) gene or single nucleotide polymorphisms (SNPs) in the puroindoline b (Pinb) gene results in hard grain texture. So far, nine Pina alleles (Pina-D1a-Pina-D1b, Pina-D1k-Pina-D1q) and seventeen Pinb alleles (Pinb-D1a-Pinb-D1g, Pinb-D1p-Pinb-D1ab) have been identified in bread wheat. The major Pina and Pinb alleles identified in hard wheat cultivars are Pina-D1b. Pinb-D1b, Pinb-D1c and Pinb-D1d. In this study, a three-primer PCR system was employed to develop nine co-dominant STS markers for genotyping Pina-D1a and Pina-D1b, whereas temperature-switch (TS) PCR was used to develop six co-dominant SNP markers for genotyping the Pinb-D1a, Pinb-D1b, Pinb-D1c and Pinb-D1d alleles. These STS and TS-PCR markers were used to verify the grain hardness genotype of 100 wheat cultivars. The reliability and genotyping accuracy of TS-PCR markers were confirmed through sequencing of PCR products and a comparison with previously published results. Therefore, STS and TS-PCR markers offer a simple, cost-effective and reliable method for high-throughput genotyping Pina and Pinb alleles to select grain hardness in wheat quality breeding programs and for wheat market classification.

• 出版日期2011-5