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作物学报 ›› 2007, Vol. 33 ›› Issue (10): 1703-1710.

• 研究论文 • 上一篇    下一篇

小麦品质性状分子标记多重PCR体系的建立

张晓科1,2, 夏先春2, 王忠伟2 , 万映秀3, 张平治3, 何心尧2, 杨燕2, 何中虎2,4,*   

  1. 1 西北农林科技大学农学院,陕西杨凌 712100;2 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;3 安徽省农业科学院作物研究所,安徽合肥 230031;4 CIMMYT中国办事处,北京100081
  • 收稿日期:2006-12-25 修回日期:1900-01-01 出版日期:2007-10-12 网络出版日期:2007-10-12
  • 通讯作者: 何中虎

Establishment of Multiplex-PCR for Quality Traits in Common Wheat

ZHANG Xiao-Ke 1,2, XIA Xian-Chun 2, WANG Zhong-Wei2, WAN Ying-Xiu 3, ZHANG Ping-Zhi3, HE Xin-Yao2, YANG Yan2, HE Zhong-Hu 2,4,*   

  1. 1 College of Agronomy, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100, Shaanxi; 2 National Wheat Improvement Center, Institute of Crop Sciences /National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, Beijing 100081; 3Crop Research Institute, Anhui Academy of Agricultural Sciences, Hefei 230031, Anhui; 4 CIMMYT-China Office, c/o Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2006-12-25 Revised:1900-01-01 Published:2007-10-12 Published online:2007-10-12
  • Contact: HE Zhong-Hu

摘要:

小麦高分子量谷蛋白亚基组成、1B/1R易位、籽粒硬度、直链淀粉含量和穗发芽抗性等性状与加工品质密切相关,建立相关性状的多重PCR体系在小麦品质分子育种中具有重要意义。本研究利用现有品质性状基因的分子标记,建立了适合不同品质类型品种评价和分子聚合育种的3个多重PCR体系,并用已知基因的品种(系)进行验证。多重PCR体系Ⅰ包括ω-secalin(1B/1R)、Vp1B3和Pinb-D1b基因的分子标记检测,可用于一般的品质检测;体系Ⅱ包括ω-secalin、Ax2*、Bx17和Dx5 基因的检测,可望用于强筋小麦品种的选育;体系Ⅲ包括Wx-A1、Wx-B1和Wx-D1位点的检测,可用于淀粉品质或糯小麦的选育。每个体系内的引物之间不存在相互抑制作用和错配,检测品种(系)的结果可靠、重复性好,成本低。3个多重PCR体系用于小麦品质育种的亲本评价和杂交后代优质基因的聚合,将会提高优质专用小麦品种评价和选育的效率。

关键词: 普通小麦, 品质性状, 多重PCR, 分子标记辅助育种

Abstract:

Wheat (Triticum aestivum L.) quality properties are strongly affected by the compositions of high-molecular-weight glutenin subunits, kernel hardness, amylase content, pre-harvest sprouting tolerance and presence or absence of 1B/1R translocation. It is very important to develop multiplex PCR for wheat quality improvement in molecular marker assisted breeding to reduce the cost and improve the efficiency. Three types of multiplex PCRs were developed and validated with 13, 30, and 11 Chinese wheat cultivars and advanced lines with known genes, respectively. The first multiplex PCR was used to simultaneously detect genes ω-secalin (1B/1R), Vp1B3, and Pinb-D1b for improving wheat processing quality. The second one was to detect the genes ω-secalin, Ax2*, Bx17, and Dx5 for improving gluten quality and bread making quality. The third multiplex PCR included three markers for Wx-7A, Wx-4A, and Wx-7D to improve starch quality and breed waxy wheat cultivars. The genotypes of all tested wheat cultivars and advanced lines identified by three multiplex PCRs were in agreement with those detected by other methods. The primer-primer interactions in each multiplex PCR were not found. Genomic DNAs extracted from both wheat kernels and leaves were feasible for three multiplex PCR amplifications. The three multiplex PCRs were highly effective in the test of Chinese wheat cultivars, demonstrating good repeatability and low cost for the evaluation of wheat quality properties in wheat breeding program.

Key words: Triticum aestivum L., Quality trait, Multiplex PCR, Molecular marker-assisted breeding

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