新疆小麦品种高分子量谷蛋白亚基及相关品质基因的分子标记检测

新疆小麦品种高分子量谷蛋白亚基及相关品质基因的分子标记检测

作物学报 ACTA AGRONOMICA SINICA 2009, 35(4): 647 661

ISSN 0496-3490; CODEN TSHPA9

/zwxb/

E-mail: xbzw@

DOI: 10.3724/SP.J.1006.2009.00647

新疆小麦品种高分子量谷蛋白亚基及相关品质基因的分子标记检测

芦 静1,2 何中虎2,3,* 夏先春2 吴新元1 李 冬1 曹俊梅1

1

新疆农业科学院粮食作物研究所, 新疆乌鲁木齐830000; 2中国农业科学院作物科学研究所/国家小麦改良中心, 北京100081; 3

CIMMYT中国办事处, 北京100081

摘 要: 高分子量麦谷蛋白亚基、1B·1R易位系、多酚氧化酶(PPO)活性及黄色素含量等基因对小麦加工品质有重要影响, 准确、快速鉴定这些基因对品质改良有重要意义。本研究对新疆当地及国内外引进的321份小麦品种进行SDS-PAGE分析, 利用特异性分子标记对高分子量谷蛋白亚基中的Dx5、Bx7、By8、By9亚基、1B·1R易位系、PPO和黄色素含量基因进行鉴定, 进一步验证分子标记检测的可靠性和准确性, 为优质小麦分子标记辅助育种提供材料和方法。SDS-PAGE分析表明, 供试材料有21种亚基类型, 其中Glu-A1位点有3种类型, 以null为主; G1u-B1位点有10种类型, Bx7+By8和Bx7+By9亚基占主导地位; Glu-D1位点有8种类型, Dx2+Dy12和Dx5+Dy10占主导地位。分子标记检测表明, Dx5亚基的频率为38.3%, Bx7亚基的频率为85.7%, By8亚基的频率为38.9%, Bx9亚基的频率为42.7%; 特异性PCR标记扩增与SDS-PAGE鉴定结果的吻合率分别为97.2%、98.4%、93.4%和97.2%。在新疆当地品种、其他国内品种和国外品种中, 1B·1R易位系材料的频率分别为22.0%、31.5%和25.0%, Psy-A1b的频率分别为9.0%、10.8%和5.4%, Ppo-A1b的频率分别为38.0%、43.8%和45.7%, Ppo-D1a的频率分别为48.0%、66.9%和40.2%。同时含Ppo-A1b和Ppo-D1a的材料有74份, 占23.0%。本试验中采用的基因特异性标记重复性好、准确率高, 可有效用于小麦品质分子标记辅助选择。

关键词: 普通小麦; 高分子谷蛋白亚基; SDS-PAGE; 品质基因; 分子标记

Characterization of Xinjiang Local and Intoduced Wheat Germplasm for High Molecular Weight Glutenin Subunits and Quality-Related Genes with Molecu-lar Markers

LU Jing1,2, HE Zhong-Hu2,3,*, XIA Xian-Chun2, WU Xin-Yuan1, LI Dong1, and CAO Jun-Mei1

1

Institute of Cereal Crops, Xinjiang Academy of Agricultural Sciences, Urumqi 830000, China; 2 Institute of Crop Sciences / National Wheat

Improvement Center, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 3 CIMMYT China Office, Beijing 100081, China

Abstract: High molecular weight glutenin subunits (HMW-GS), 1B·1R translocation, polyphenol oxidase (PPO) activities, and yellow pigment content are mostly correlated with the processing quality of common wheat (Triticum aestivum L.). An accurate and fast characterization of these genes is of great importance in the improvement of wheat quality. In this study, a total of 321 wheat genotypes, including 100 Xinjiang local wheat cultivars, 130 introductions from other provinces of China, and 91 introduc-tions from other countries, were analyzed by SDS-PAGE method. In addition, the functional markers of Dx5, Bx7, By8, By9, 1B·1R, PPO16, PPO18, PPO29, and YP7A were used to detect their allelic variations. Twenty-one subunit combinations were found according to the SDS-PAGE data, three types at Glu-A1 locus with null as a major subunit, 10 types at Glu-B1 locus with Bx7+By8 and Bx7+By9 as major subunits, and eight types at Glu-D1 locus with Dx2+Dy12 and Dx5+Dy10 as major subunits. The frequencies of the Dx5, Bx7, By8, and Bx9 subunits revealed by functional markers were 38.3%, 85.7%, 38.9%, and 42.7%, respectively, with 97.2%, 98.4%, 93.4%, and 97.2% of consistency with SDS-PAGE results, respectively. Eighty-six genotypes had 1B·1R translocations, with 22.0% in Xinjiang local cultivars, 31.5% in the introductions from other provinces of China, and 25.0% in those from other countries, respectively. The frequencies of Psy-A1b detected by YP7A marker were 9.0%, 10.8%, and 5.4% among three group genotypes, respectively. The PPO18 marker for the Ppo-A1 locus yielded Ppo-A1b allele with frequen-cies of 38.0%, 43.8%, and 45.7% among three group genotypes, respectively. The Ppo-D1a allele frequencies were 48.0%, 66.9%,

本研究由中国科学院“西部之光”计划项目, 国家高技术研究发展计划(863计划)重点项目(2006AA10Z1A7和2006AA100102), 新疆维吾尔自通讯作者(Corresponding author): 何中虎, E-mail: zhhe@; Tel: 010-82105691; Fax: 010-82108547

治区重大科技专项(200731132-1)资助。

*

Received(收稿日期): 2008-08-13; Accepted(接受日期): 2008-10-25.

新疆小麦品种高分子量谷蛋白亚基及相关品质基因的分子标记检测

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作 物 学 报 第35卷

and 40.2% among three group genotypes, respectively, according to the PPO16 and PPO29 markers at Ppo-D1 locus. However, only 74 genotypes contained Ppo-A1b and Ppo-D1a alleles at both loci, accounting for 23.0% of the genotypes. The functional markers applied in this study were repeatable, accurate and stable, and can be effectively used in wheat quality breeding. Keywords: Common wheat; HMW-GS; SDS-PAGE; Processing quality; Molecular markers

小麦面筋质量的优劣与高分子量谷蛋白亚基(HMW-GS)密切相关[1-3]。不同高分子量谷蛋白亚基对品质的贡献不同, 具备高分子量谷蛋白亚基组合1或2*、7+8/7+9或17+18、5+10的基因型具有较好的面包品质。过去主要依据亚基在聚丙烯酰胺凝胶(SDS-PAGE)中的迁移率来鉴定高分子量谷蛋白亚基的组成, 由于电泳图谱上亚基的迁移率与分子量并不总是一致, 从而混淆分子量不同而迁移率相似的亚基, 因此在精确度上有待进一步提高。利用分子标记进行高分子量谷蛋白亚基鉴定, 克服了SDS-PAGE方法的局限性, 在不同生育期都可以取材检测, 不必破坏种子, 可以快速大量地检测有关基因, 因而受到广泛重视。目前, 已获得了部分高分子量谷蛋白亚基基因的分子标记[4-8], 如Ax1、Ax2*、Bx7、By8、By9、Bx17、Dx5和Bx20等, 为分子标记辅助育种提供了技术保障。1B·1R易位系在中国小麦育种和生产中发挥了重要作用[9], 但这也是导致我国小麦加工品质(尤其是面包烘烤品质)较差的主要原因之一[10-11]。为了快速检测1B·1R易位系, Francis等[12]开发了特异性基因标记, 并得到广泛应用。

面粉及其制品的色泽是衡量小麦品质的一项重要指标。多酚氧化酶活性(PPO)、黄色素含量、蛋白质含量和脂肪氧化酶(lipoxygenase, LOX)活性等都影响面制品颜色。高多 …… 此处隐藏：4545字，全部文档内容请下载后查看。喜欢就下载吧 ……