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

doi:10.3724/SP.J.1005.2010.00613

遗传 ›› 2010, Vol. 32 ›› Issue (6): 613-624.doi: 10.3724/SP.J.1005.2010.00613

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

王林海¹, 周敏², 李慧玲³, 何中虎^{1, 4}, 夏先春¹

1. 中国农业科学院作物科学研究所, 国家小麦改良中心, 国家农作物基因资源与基因改良重大科学工程, 北京100081;
2. 北京农学院, 北京102206;
3. 河北省廊坊市种子管理站, 廊坊 065000;
4. 国际玉米小麦改良中心(CIMMYT)中国办事处, 北京 100081

收稿日期:2009-10-15 修回日期:2009-12-31 出版日期:2010-06-20 发布日期:2010-05-24
通讯作者: 周敏;夏先春 E-mail:zmskx@yahoo.com.cn;xiaxianchun@caas.net.cn
基金资助:
国家高技术研究发展计划项目(863计划)(编号：2006AA10Z1A7, 2006AA100102)和国家自然科学基金项目(编号：30830072)资助

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

WANG Lin-Hai¹, ZHOU Min², LI Hui-Ling³, HE Zhong-Hu^{1, 4}, XIA Xian-Chun¹

1. Institute of Crop Science, National Wheat Improvement Center/The National Key Facility for Crop Gene Resources and Genetic Im-provement, Chinese Academy of Agricultural Sciences (CAAS), Beijing 100081, China;
2. Beijing Agricultural College, Beijing 102206, China;
3. The Department of Seed Administration, Langfang 065000, China;
4. International Maize and Wheat Improvement Center (CIMMYT) China Office, Beijing 100081, China

Received:2009-10-15 Revised:2009-12-31 Online:2010-06-20 Published:2010-05-24
Contact: ZHOU Min;XIA Xian-Chun E-mail:zmskx@yahoo.com.cn;xiaxianchun@caas.net.cn

摘要/Abstract

摘要：

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。

关键词: 普通小麦, 小麦近缘种, 低分子量麦谷蛋白亚基, Glu-B3, 系统发育

Abstract:

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.

Key words: Triticum aestivum L., wheat related species, LMW-GS, Glu-B3, phylogenetics

王林海，周敏，李慧玲，何中虎，夏先春. 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[J]. 遗传, 2010, 32(6): 613-624.

WANG Lin-Hai, ZHOU Min, LI Hui-Ling, HE Zhong-Hu, JIA Xian-Chun. Cloning and phylogenetic analysis of low-molecular-weight glutenin subunit genes at Glu-B3 locus in common wheat relative species[J]. HEREDITAS, 2010, 32(6): 613-624.

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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

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

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