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中国农学通报 ›› 2012, Vol. 28 ›› Issue (1): 195-200.doi: 10.11924/j.issn.1000-6850.2011-2147

所属专题: 生物技术 园艺

• 林学 园艺 园林 • 上一篇    下一篇

番茄黄化卷叶病毒病抗病基因Ty-1的CAPS标记建立

韩璀璇 宋建军 王琳珊 田鹏 仇燕   

  • 收稿日期:2011-07-25 修回日期:2011-10-17 出版日期:2012-01-05 发布日期:2012-01-05
  • 基金资助:

    河北省自然科学基金项目;河北省科技支撑计划项目

Establishment of CAPS Molecular Marker for Ty-1 Gene Resistant to Tomato Yellow Leaf Curl Disease

  • Received:2011-07-25 Revised:2011-10-17 Online:2012-01-05 Published:2012-01-05

摘要:

本研究旨在筛选获得与番茄黄化卷叶病毒抗病基因Ty-1紧密连锁的分子标记,为番茄抗病育种提供技术支撑。根据与抗病基因Ty-1紧密连锁的RFLP标记序列设计特异引物,以抗病杂合体(Ty-1/ty-1)、抗病纯合体(Ty-1/Ty-1)和感病纯合体(ty-1/ty-1)材料提取的DNA为模板,进行PCR扩增,而后经不同的核酸内切酶酶切处理,筛选获得与Ty-1基因紧密连锁的CAPS标记。结果显示从4个标记中筛选获得了2个稳定可靠的CAPS标记,即TG97和Mi23。TG97标记在抗病杂合体产生398 bp、303 bp和95 bp 3个特异片段,抗病纯合体产生303 bp和95 bp 2个特异片段,感病纯合体产生398 bp一个特异片段。Mi23标记在抗病杂合体产生402 bp和354 bp 2个特异片段,抗病纯合体产生402 bp一个特异片段,感病纯合体产生354 bp一个特异片段。研究结果表明TG97和Mi23这2个CAPS标记均为共显性标记,可用于番茄抗病育种的辅助选择中。

关键词: 密度, 密度, 施氮量, 红土晒烟

Abstract:

The study was designed to develop and obtain molecular markers tightly linked to Ty-1 gene resistant to tomato yellow leaf curl virus, in order to affording technical supports for tomato disease resistance breeding. In this study, resistant heterozygotes (Ty-1/ty-1), resistant homozygotes (Ty-1/Ty-1) and susceptible homozygotes (ty-1/ty-1) were used as experimental materials. Based on the published RFLP markers tightly linked to Ty-1, four pairs of specific primers were designed for PCR amplification. PCR products were then digested with different restriction enzymes to develop CAPS molecular markers. As a result, two steady-going and available CAPS molecular markers were found, i.e. TG97 and Mi23. TG97 marker showed three restriction fragments (398 bp, 303 bp and 95 bp) with resistant heterozygous materials, two restriction fragments (303 bp and 95 bp) with resistant homozygous materials and only 398 bp fragment with susceptible homozygous materials. Mi23 marker showed two restriction fragments (402 bp and 354 bp) with resistant heterozygous materials, only 402 bp fragments with resistant homozygous materials and only 354 bp fragments with susceptible homozygous materials. These findings indicated that the TG97 and Mi23 were co-dominant CAPS markers, which could be used in tomato breeding for disease resistance.

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