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

¹Institute of Crop Breeding and Cultivation, Chinese Academy of Agricultural Science, Beijing, 100081, China. ²Corresponding authors. E-mail: panzhm@mail.caas.net.cn; jzhu@chguenther.com ³Dept. of Cereal & Food Sciences, North Dakota State University, Fargo, ND 58105. ⁴Grain Science and Industry Program, Zhengzhou Institute of Technology, Zhengzhou 450052, China.

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.