State Key Laboratory of Plant Cell and Chromosome Engineering, National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 1 West Beichen Road, Chaoyang District, Beijing, China.
Theor Appl Genet. 2011 May;122(8):1503-16. doi: 10.1007/s00122-011-1550-7. Epub 2011 Feb 23.
Low-molecular-weight glutenin subunits (LMW-GSs) play an important role in determining the bread-making quality of bread wheat. However, LMW-GSs display high polymorphic protein complexes encoded by multiple genes, and elucidating the complex LMW-GS gene family in bread wheat remains challenging. In the present study, using conventional polymerase chain reaction (PCR) with conserved primers and high-resolution capillary electrophoresis, we developed a new molecular marker system for identifying LMW-GS gene family members. Based on sequence alignment of 13 LMW-GS genes previously identified in the Chinese bread wheat variety Xiaoyan 54 and other genes available in GenBank, PCR primers were developed and assigned to conserved sequences spanning the length polymorphism regions of LMW-GS genes. After PCR amplification, 17 DNA fragments in Xiaoyan 54 were detected using capillary electrophoresis. In total, 13 fragments were identical to previously identified LMW-GS genes, and the other 4 were derived from unique LMW-GS genes by sequencing. This marker system was also used to identify LMW-GS genes in Chinese Spring and its group 1 nulli-tetrasomic lines. Among the 17 detected DNA fragments, 4 were located on chromosome 1A, 5 on 1B, and 8 on 1D. The results suggest that this marker system is useful for large-scale identification of LMW-GS genes in bread wheat varieties, and for the selection of desirable LMW-GS genes to improve the bread-making quality in wheat molecular breeding programmes.
高分子量谷蛋白亚基(LMW-GSs)在决定面包小麦的面包制作质量方面起着重要作用。然而,LMW-GSs 表现出高度多态性的蛋白质复合物,由多个基因编码,阐明面包小麦中复杂的 LMW-GS 基因家族仍然具有挑战性。在本研究中,我们使用常规聚合酶链反应(PCR)与保守引物和高分辨率毛细管电泳,开发了一种新的分子标记系统,用于鉴定 LMW-GS 基因家族成员。基于先前在我国小麦品种小偃 54 中鉴定的 13 个 LMW-GS 基因和 GenBank 中可用的其他基因的序列比对,设计了 PCR 引物,并分配给跨越 LMW-GS 基因长度多态性区域的保守序列。PCR 扩增后,使用毛细管电泳检测小偃 54 中的 17 个 DNA 片段。总共检测到 13 个与先前鉴定的 LMW-GS 基因相同的片段,另外 4 个片段是通过测序从独特的 LMW-GS 基因衍生而来的。该标记系统还用于鉴定中国春及其 1 组缺四体系。在所检测的 17 个 DNA 片段中,4 个位于 1A 染色体上,5 个位于 1B 染色体上,8 个位于 1D 染色体上。结果表明,该标记系统可用于大规模鉴定面包小麦品种中的 LMW-GS 基因,并用于选择理想的 LMW-GS 基因,以提高小麦分子育种计划中的面包制作质量。