Abstract
Kernel hardness is mainly controlled by one major genetic locus on the short arm of chromosome 5D in bread wheat. Twelve Chinese and CIMMYT wheat cultivars were characterized for the deletion region of Pina-D1b genotype and developing a novel STS marker for this allele. PCR and SDS-PAGE were used to confirm the Pina-D1b genotype, and then 20 pairs of primers were designed to amplify the fragment including deletion region in Pina-D1b genotype by primer walking strategy. An STS marker Pina-N spanning deletion region in Pina-D1b was developed and sequencing results showed that all of 10 Pina-D1b genotypes uniformly possessed a 15,380 bp deletion in comparison with that of Chinese Spring with wild type. This study provided an alternative method to exam Pina-D1b by molecular marker and will accelerate identification of puroindoline alleles in bread wheat.
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