中国农业科学 ›› 2008, Vol. 41 ›› Issue (3): 643-653 .doi: 10.3864/j.issn.0578-1752.2008.03.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

多重PCR的建立及黄淮麦区主要品种品质相关基因的鉴定

万映秀,张晓科,夏先春,张平治,何中虎   

  1. 安徽省农业科学院作物研究所/安徽省农作物品质改良重点实验室
  • 收稿日期:2007-01-09 修回日期:1900-01-01 出版日期:2008-03-10 发布日期:2008-03-10
  • 通讯作者: 张平治,何中虎

Identification of Major Quality Genes in Cultivars from Yellow and Huai River Valley Wheat Region by Multiplex PCR

  1. 安徽省农业科学院作物研究所/安徽省农作物品质改良重点实验室
  • Received:2007-01-09 Revised:1900-01-01 Online:2008-03-10 Published:2008-03-10

摘要: 【目的】小麦加工品质由多个位点控制,而每个位点又有多个等位基因控制。建立品质性状多重PCR反应体系是提高分子标记辅助选择效率及降低成本的一种重要措施。【方法】选择影响小麦品质性状重要基因的分子标记,即高分子量谷蛋白亚基基因标记Ax2*、Bx14、Bx17和Dx5;低分子量谷蛋白亚基基因标记Glu-A3 d;糯蛋白亚基Wx-B1基因标记BDFL-BRD和Wx-D1基因标记MAG269;1BL/1RS易位标记ω-sec及多酚氧化酶(PPO)活性基因标记PPO18。根据优质面包、优质面条亚基组成要求及引物的退火温度,建立3个多重PCR反应体系PCR-Ⅰ(Ax2*/Bx17/Dx5)、PCR-Ⅱ(BDFL-BRD/MAG269/PPO18)和PCR-Ⅲ(Bx14/Glu-A3d/ω-sec)。【结果】用构建的3个多重PCR对141份黄淮麦区小麦品种加工品质相关基因的分布情况进行了检测。结果表明Ax2*、Bx14、Bx17具有较低的分布频率,分别为4.3%、7.1%、1.4%;Dx5和Glu-A3 d分布频率相对较高,分别为17.7%和27.7%;而1BL/1RS易位和低PPO活性(扩增片段为876 bp)的材料分别占44.0%和51.8%;Wx-B1缺失材料占4.3%。在供试的141份材料中已有80份进行了高、低分子量谷蛋白亚基和黑麦碱的SDS-PAGE电泳检测,与本研究建立的多重PCR检测结果完全一致。【结论】建立的多重PCR体系可以准确、稳定、高效地检测9个小麦品质性状的基因组成。

关键词: 普通小麦, 加工品质, 多重PCR

Abstract: Abstract: 【Objective】Development of multiplex PCR for wheat quality traits is very important to improve the efficiency and reduce the cost in molecular marker assisted breeding. 【Method】 In the present study, according to genes controlling for pan bread and Chinese noodle quality, and the annealing temperatures of different markers, the molecular markers for important quality trait genes, i.e., high-molecular-weight glutenin subunit (HMW-GS) genes Ax2*, Bx14, Bx17 and Dx5, and low-molecular-weight glutenin subunit (LMW-GS) gene Glu-A3d, as well as the marker BDFL-BRD for Wx-B1, MAG269 for Wx-D1, ω-sec for 1BL/1RS translocation and PPO18 for polyphenol oxidase(PPO)activity, were selected and three combinations of molecular markers Ax2*/Bx17/Dx5, Bx14/Glu-A3d/ω-sec and BDFL-BRD/MAG269/PPO18 were tested. 【Result】Results indicated that the three multiplex PCRs, PCR-I (Ax2*/Bx17/Dx5), PCR-II (Bx14/Glu-A3d/ω-sec) and PCR-III (BDFL-BRD/MAG269/PPO18) can be used to identify the genes presented in wheat cultivars. A total of 141 wheat cultivars were evaluated with the multiplex PCRs, and the frequencies of HMW-GS Ax2*, Bx14, Bx17, Dx5 and LMW-GS Glu-A3d accounted for 4.3%, 7.1%, 1.4%, 17.7% and 27.7%, respectively. Frequencies of 1BL/1RS translocation and low polyphenol oxidase (PPO) activity genetypes were 44.0% and 51.8%, respectively. Wx-B1 null type accounted for 4.3%. Data obtained by SDS-PAGE for HMW-GS, LMW-GS and secalin for 80 genotypes correspond very well with those from the multiplex PCRs. 【Conclusion】This indicated that the multiplex PCRs developed can be steadily and efficiently used to detect the nine genes for the improvement of wheat quality in molecular marker assisted breeding.