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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (15): 38-43.

所属专题： 生物技术； 小麦

• 生物技术科学 • 上一篇    下一篇

利用SDS-PAGE和AS-PCR标记鉴定簇毛麦HMW-GS基因

刘守斌 梁荣奇   

  • 收稿日期:2010-02-20 修回日期:2010-03-26 出版日期:2010-08-05 发布日期:2010-08-05
  • 基金资助:

    国家自然科学基金项目;北京市自然科学基金项目

Analysis on HMW-GS genes in Dasypyum villosum by SDS-PAGE and Allelic Specific (AS) -PCR Markers

  • Received:2010-02-20 Revised:2010-03-26 Online:2010-08-05 Published:2010-08-05

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摘要：

本研究通过SDS-PAGE和AS-PCR标记对簇毛麦HMW-GS进行鉴定，对于快速鉴别导入到普通小麦中的簇毛麦HMW-GS和改良普通小麦品质具有重要意义。利用SDS-PAGE对中国春、钱尼、簇毛麦、6个中国春-簇毛麦二体附加系、普通小麦-簇毛麦3V异代换系进行HMW-GS组成分析，进一步确定簇毛麦HMW-GS基因位于1V染色体。根据已发表的普通小麦HMW-GS基因序列同源性比较结果，设计了3对分别扩增普通小麦x-型亚基基因、y-型亚基基因以及所有HMW-GS基因的特异引物。经过对簇毛麦HMW-GS基因进行扩增发现，这3个AS-PCR标记都能很好地鉴别簇毛麦HMW-GS编码基因，并从分子水平上把簇毛麦HMW-GS基因定位于簇毛麦1V染色体上。

关键词: 抗旱复合肥, 抗旱复合肥, 玉米, 水分利用率, 氮素利用率

Abstract:

Sodium Dodecyl Sulphate -PolyAcrylamide Gel Electrophoresis (SDS-PAGE) and three Allelic Specific PCR (AS-PCR) markers were used in this paper to screen and confirm the High-molecular-weight glutenin subunits (HMW-GS) of D. villosum. The results of analysis on HMW-GS of CS, Cheyenny, D. villosum, six addition lines of D. villosum chromosome in CS and substitution line 3V of D. villosum chromosome in T. aestivum by SDS-PAGE indicated that genes encoding for HMW-GS were located in D. villosum chromosome 1V. Three AS-PCR markers, which could amplify x-type HMW glutenin genes, y-type HMW glutenin genes and all HMW glutenin genes of T. aestivum respectively, were synthesized on the basis of all published sequences of HMW-GS genes from T. aestivum. The results of amplification in D. villosum and other materials above showed that all of three markers could be used for characterization of HMW glutenin genes in D. villosum. Therefore, the gene encoding for HMW-GS of D. villosum was located in chromosome 1V in molecular level.