| 摘要: |
| 【目的】构建白粉菌诱导下中国野生毛葡萄“商-24”的抑制消减文库，获得抗白粉病差异表达基因EST序列。【方法】应用抑制消减杂交（Suppression subtractive hybridization，SSH）技术，构建白粉菌诱导下高抗白粉病的中国野生毛葡萄“商-24”叶片SSH cDNA文库,通过生物信息学方法对文库中的EST序列进行分析。【结果】成功构建了白粉菌诱导的中国野生毛葡萄“商-24”抑制消减cDNA文库，对消减文库测序后获得了200个EST序列，大小为300～1 000 bp；BLAST分析表明，其中19个EST序列与已知功能基因序列同源性很高，分别参与了植物的防卫反应、胁迫反应、细胞解毒和信号转导等。【结论】获得了葡萄抗白粉病的EST序列，这为进一步克隆抗葡萄白粉病基因、探明其抗病分子机理奠定了基础。 |
| 关键词: 毛葡萄 抗白粉病 消减文库 EST |
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| 基金项目:国家自然科学基金项目（30671446，30871701）；教育部新世纪优秀人才支持计划及国家葡萄现代产业技术体系项目（nycytx-30-zp-06） |
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| Construction and EST analysis of a suppressive subtraction cDNA library of Chinese Wild Vitis quinquangularis inoculated with powdery mildew |
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| Abstract: |
| 【Objective】The study was to construct a suppressive subtraction cDNA library in Chinese Wild Vitis quinquangular Shang-24 inoculated with powdery mildew and to obtain disease resistance EST sequence related to powny mildew.【Method】 Suppression subtractive hybridization was employed to construct the SSH cDNA library of Chinese Wild V.quinquangular Shang-24 with powdery mildew,using bioinformatics method to analyses these ESTs.【Result】 A suppressive subtraction cDNA library of Chinese Wild V.quinquangular inoculated with powdery mildew were constructed successfully,200 ESTs were obtained from the SSH library,the fragments were 300-1 000 bp in length.BLAST analysis revealed that 19 ESTs had high homology with known genes in GenBank involved in defense response,stress reaction,disease defense detoxification and signal transduction.【Conclusion】 The ESTs of disease resistant obtained from SSH library is the foundation for further cloning grape powdery mildew resistance genes and understanding the molecular mechanism of disease resistant. |
| Key words: Vitis quinquangular resistance to powdery mildew subtracted cDNA library EST |
