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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (12): 194-200.doi: 10.11924/j.issn.1000-6850.2012-2479

所属专题： 小麦

• 植物保护 农药 • 上一篇    下一篇

NBS profiling技术在小麦抗叶锈病研究中的应用初探

梁丽然 王珅 高琳 王海燕 刘大群   

  • 收稿日期:2012-07-13 修回日期:2012-08-04 出版日期:2013-04-25 发布日期:2013-04-25
  • 基金资助:
    河北省自然科学基金;转基因生物新品种重大科技专项

Preliminary Study of Wheat Leaf Rust Disease using NBS profiling

  • Received:2012-07-13 Revised:2012-08-04 Online:2013-04-25 Published:2013-04-25

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摘要： 为了研究NBS profiling技术在小麦(Triticum aestivum L.)抗叶锈病基因中应用的可行性，本研究选用15个小麦抗叶锈病近等基因系和感病亲本Thatcher为材料，建立NBS profiling体系，并进行多态性分析；同时选取含有成株抗叶锈病基因Lr35的近等基因系TcLr35和Thatcher为材料，进行cDNA NBS profiling，从转录水平上寻找与目的基因Lr35相关的RGA片段。研究结果表明：基因组NBS profiling中所选的16对酶/引物组合中MseⅠ/NBS2、MseⅠ/NBS5和MseⅠ/NBS7均得到了较好扩增结果，多态性高，多态性检出率分别为26.4%、26.4%和23.5%；cDNA NBS profiling中共筛选了52对酶/引物组合进行差异片段的扩增，有多对组合能成功揭示抗感间差异。NBS profiling技术可应用于小麦叶锈病抗病基因的研究中，为该技术在小麦抗叶锈病基因筛选及分子标记研究中的应用奠定了基础。

关键词: 诊断分析, 诊断分析

Abstract: In order to study the feasibility of NBS profiling in leaf rust resistance gene, fifteen near-isogenic lines (NILs) and the susceptible parent Thatcher were selected as materials in the study. The NBS profiling system was established in wheat successfully and the polymorphism was analyzed. In the meanwhile, the TcLr35 wheat containing wheat leaf rust resistance gene Lr35 and Thatcher was used for the cDNA NBS profiling in order to uncover the RGA fragments related to Lr35 gene in the transcriptional level. The experimental results showed that the combination of MseⅠ/NBS2, MseⅠ/NBS5, MseⅠ/NBS7 could amplify the polymorphic bands and the rate of polymorphism was 26.4%, 26.4%, 23.5% respectively among the sixteen combinations of enzyme/primer; Fifty-two combinations of enzyme/primer were used to screen the specific fragment using cDNA NBS profiling between resistant and susceptible materials; several combinations could reflect the difference in TcLr35 and Thatcher. These results indicated that NBS profiling could be used for the study of wheat leaf rust resistance gene, which was the foundation to screen the wheat leaf rust resistance gene and the molecular markers..