中国春小麦籽粒硬度puroindoline基因等位变异检测

Allelic Variation of Puroindoline Genes in Chinese Spring Wheats

【目的】研究中国不同麦区春小麦籽粒硬度基因型分布规律,为小麦品质改良和各麦区间资源交流提供依据。【方法】以中国7个省、自治区的140份春小麦主栽品种、高代品系和主要亲本为材料,采用单籽粒谷物硬度测试仪、特异引物的PCR扩增、PCR产物的酶切(PvuII内切酶)和改进的SDS-PAGE凝胶电泳对其SKCS硬度及其基因型进行鉴定。【结果】中国春麦区以硬质类型为主,共有野生型、Pina-D1b、Pinb-D1b、Pinb-D1c(先前未曾在中国冬麦区发现)和Pinb-D1p共5种类型。其中,Pinb-D1b在硬质麦占据主导地位,为51.5%;其次是Pina-D1b,为37.9%;Pinb-D1c和Pinb-D1p类型分别为4.9%和5.8%。后二者分布具有明显的地域性,Pinb-D1c主要分布在青海,Pinb-D1p则主要分布在甘肃。各puroindoline突变型SKCS硬度值显著高于野生型,Pina-D1b和Pinb-D1c类型SKCS硬度值显著高于Pinb-D1b和Pinb-D1p类型。【结论】本试验明确了puroindoline基因在中国春麦区的分布规律,总体上虽然以Pinb-D1b类型为主,但Pina-D1b类型分布也较为广泛,尤其在黑龙江等地区,因其对品质加工有一定的负面影响,建议该类型分布比例较高的地区多引进一些其它相对优良的基因型,如Pinb-D1b类型等。

[Objective] Study of wheat kernel hardness is helpful to improving quality and introducing materials in Chinese spring wheat regions. [Method] In total, 140 Chinese spring wheat cultivars, advanced lines, and breeding parents from seven provinces in China were examined by a Single Kernel Characterization System （SKCS）, STS （sequence tagged site） marker, site-specific cleavage with Pvull, SDS-PAGE and sequencing to understand the distribution of grain hardness and its puroindoline alleles. [Result] The results indicated that wild type, Pina-Dlb, Pinb-Dlb, Pinb-Dlc （absent in Chinese winter wheats） and Pinb-Dlp were present in Chinese spring wheats, and Pinb-Dlb was the most popular genotype with a frequency of 51.5% in hard spring wheat. The percentages ofPina-Dlb, Pinb-Dlc and Pinb-Dlp were 37.9%, 4.9% and 5.8% in hard spring wheats, respectively Pinb-Dlc and Pinb-Dlp are mainly distributed in Qinghai and Gansu provinces, respectively. In addition, SKCS hardness indexes of mutation genotypes are significantly higher than that of wild type and SKCS hardness indexes of Pina-Dlb and Pinb-Dlc are significantly higher than those of Pinb-Dlb and Pinb-Dlp. [ Conclusion] This study characterized the distribution ofpuroindoline alleles in Chinese spring wheat and provided the fact that Pinb-Dlb are the most popular genotype in Chinese spring wheat region. Also, Pina-Dlb was very prevalent, especially in Heilongiiang, suggesting that some relatively superior genotypes should be introduced to regions with high percentages of Pina-Dlb due to its inferior quality. This study could provide useful information for improvement of wheat quality in Chinese spring wheat region.