Characterization of Genes Encoding Wheat Grain Hardness from Chinese Cultivar GaoCheng 8901

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Abstract
The puroindoline‐b (Pinb‐D1) gene from Chinese hard wheat cultivar GaoCheng 8901 (Triticum aestivum L.) was obtained using two pairs of primers designed based on the known Pinb‐D1 gene sequence and polymerase chain reaction (PCR) amplification. The PCR amplification was made using the genomic DNA of the wheat as a template and the specific fragment ≈450 bp in size was screened. The results indicated that the Pinb‐D1 gene in GaoCheng 8901 shared 99.78% and 99.32% homology in nucleotide acid sequence and amino acid sequence, respectively, compared with the Pinb‐D1 gene from hard wheat cultivars Wanser and Cheyenne. A new mutation in this Pinb‐D1 gene, different from the six known mutations in the Pinb‐D1 gene, was characterized with a change of a lysine to glutamic acid at position 45 in its protein sequence. This mutation, designated as Pinb‐D1l in this study, might contribute to the formation of grain hardness in GaoCheng 8901. The characterization of Pinb‐D1 gene would be helpful in manipulating grain hardness of wheat through genetic engineering.