利用单籽粒谷物特性测试仪和PCR扩增、酶切及DNA测序技术，结合改良的friabilin提取及电泳分析方法，对山东省431份农家品种、63份历史品种和29份当前主栽品种的籽粒硬度分布以及Pina和Pinb等位变异类型进行研究，以探讨山东小麦籽粒硬度的演变规律。农家品种、历史品种和当前主栽品种中硬质麦比例分别为75.6%、12.7%和27.6%，混合麦分别占20.4%、19.0%和13.8%，而软质麦分别占3.9%、68.3%和58.6%。农家品种中共有6种基因型，Pina-D1b/Pinb-D1a和Pina-D1a/Pinb-D1p分别占硬质麦的38%和59.6%。历史品种中有4种基因型，8份硬质麦中Pina-D1b/Pinb-D1a占37.5%，Pina-D1a/Pinb-D1b占37.5%，而Pina-D1a/Pinb-D1p只占25.0%。当前主栽品种中，8份硬质麦全部是Pina-D1a/Pinb-D1b类型。长芒透垅白等3个品种的Pinb基因的核苷酸序列发生了双突变，起始密码子下游96 bp处碱基C突变为A，而且在265 bp处发生了A碱基的缺失，将其命名为Pinb-D1aa。
Studies on the grain hardness and puroindolines alleles in wheat cultivars released in different historical periods, are helpful for breeding wheat cultivar with optimal endosperm texture. In the present study, 523 accessions from Shandong Province including 431 landraces, 63 historical cultivars and 29 current cultivars were used to evaluate the SKCS hardness and distribution of puroindoline alleles (Pins). Distribution of grain hardness differed in landraces, historical cultivars and current wheats, with 75.6%, 20.4%, and 3.9% of hard texture, and 20.4%, 19.0%, and 13.8% of mixed wheats, and 3.9%, 68.3%, and 58.6% of soft grains, respectively. Six genotypes of Pina and Pinb were present in landraces, in which Pina-D1a/Pinb-D1p and Pina-D1b/Pinb-D1b were the dominant genotypes, accounting for 38.0% and 59.6% of hard wheat, respectively. Compared with landraces, the polymorphism of Pina and Pinb was decreased in historical cultivars. Pina-D1b/Pinb-D1a, Pina-D1a/Pinb-D1b, and Pina-D1a/Pinb-D1p accounted for 37.5%, 37.5%, and 25.0% of hard wheat, respectively, whereas, Pina-D1a/Pinb-D1b was the only genotype presented in hard genotype of current cultivars surveyed. A novel Pinb allele with double mutations at the positions of 96th (C to A) and 265th (deletion of A) was found in three landraces, and was designated as Pinb-D1aa.
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