小麦近缘种低分子量麦谷蛋白亚基基因Glu-B3克隆及系统发育分析

发掘小麦近缘种低分子量麦谷蛋白基因, 可为小麦品质改良提供更多的基因资源。文章利用Glu-B3位点特异性标记LB1F/LB1R、LB2F/LB2R、LB3F/LB3R和 LB4F/LB4R, 对普通小麦B染色体组的7个可能供体近缘种, 即硬粒小麦(T. durum)、栽培二粒小麦(T. dicoccum)、野生二粒小麦(T. dicoccoides)、拟斯卑尔脱山羊草(Ae. speltoides)、高大山羊草(Ae. longissima)、西尔斯山羊草(Ae. searsii)和双角山羊草(Ae. bicornis)共20份材料进行PCR扩增, 克隆小麦近缘种中GluB3-1、GluB3-2、GluB3-3和GluB3-4基因的等位变异, 并对Glu-B3位点基因进行系统发育分析。共获得16个新等位变异, 其中GluB3-1基因的新等位变异1个, 命名为GluB3-16, 其推导氨基酸分子量为39.2 kDa; GluB3-3的新等位变异有3个, 分别命名为GluB3-35、GluB3-36和GluB3-37, 其推导氨基酸分子量为44.5 kDa(GluB3-36)或44.6 kDa(GluB3-35和GluB3-37); GluB3-4的新等位变异12个, 分别命名为GluB3-46、GluB3-47、GluB3-48、GluB3-49、GluB3-410、GluB3-411、GluB3-412、GluB3-413、GluB3-414、GluB3-415、GluB3-416和GluB3-417, 其推导氨基酸分子量变化在38.6(GluB3-414)~ 42.5 kDa(GluB3-413)之间; 16个新等位变异都包含单一的完整开放阅读框, 具有低分子量麦谷蛋白亚基的典型结构。文章进一步拓展了低分子量麦谷蛋白基因资源, 揭示不同Glu-B3基因的进化过程不完全相同, 为有效地利用小麦近缘种材料和转基因育种提供了新的基因资源。

Cloning and phylogenetic analysis of low-molecular-weight glutenin subunit genes at Glu-B3 locus in common wheat relative species

The common wheat relative species are important germplasm for wheat breeding. In the present study, novel allelic variants at Glu-B3 locus were cloned to provide gene resources for wheat quality improvement. Four Glu-B3-locus specific primer sets LB1F/LB1R, LB2F/LB2R, LB3F/LB3R, and LB4F/LB4R were employed to isolate novel allelic variants of GluB3-1, GluB3-2, GluB3-3, and GluB3-4 from seven common wheat relative species, i.e., T. durum, T. dicoccum, T. dicoccoides, Aegilops longissima, Ae. searsii, Ae. Bicornis, and Ae. speltoides, and the software MEGA 4 was used to construct a phylogenetic tree. In total, 16 novel allelic variants of GluB3-1, GluB3-3, and GluB3-4 genes were isolated from the seven common wheat relative species, designated GluB3-16, GluB3-35, GluB3-36, GluB3-37, GluB3-46, GluB3-47, GluB3-48, GluB3-49, GluB3-410, GluB3-411, GluB3-412, GluB3-413, GluB3-414, GluB3-415, GluB3-416 and GluB3-417, respectively. In detail, GluB3-16 was cloned from T. dicoccoides with LB1F/LB1R, and the molecular weight of the de-duced amino acid was 39.2 kDa. GluB3-35, GluB3-36, and GluB3-37 were isolated from T. durum and T. dicoccum with the primer set LB3F/LB3R, and the molecular weights of their deduced peptides were 44.5 kDa (GluB3-36) and 44.6 kDa (GluB3-35 and GluB3-37). The molecular weight of deduced peptides of GluB3-4 ranged from 38.6 kDa (GluB3-414) to 42.5 kDa (GluB3-413). All the 16 new allelic variants showed a single open reading frame (ORF), and their deduced amino-acid sequences had a typical sequence structure of LMW-GS. The allelic variants at Glu-B3 locus identified in com-mon wheat relative species provide potential gene resources for wheat quality breeding and gene transformation. The results suggested that these Glu-B3 genes originated from different evolution processes.