Uncinula necator is a worldwide serious fungus disease causing annual heavy lost on grapevine production. In order to get more informations on defense related EST sequences and help breeding program, we constructed an...Uncinula necator is a worldwide serious fungus disease causing annual heavy lost on grapevine production. In order to get more informations on defense related EST sequences and help breeding program, we constructed and characterized a suppression subtractive hybridization cDNA library with artificially inoculated leaves and control Chinese wild Vitis pseudoreticulata clone Baihe-35-1, which is highly resistant to powdery mildew. In the library, the length of 58 EST fragments known as putative functions varied from 130 to 800 bp, and 60% of the ESTs exhibited high similarity to known sequences in database of GenBank with BLASTX analysis. These genes were involved in stress/defense response, detoxification, signal transduction, disease defense, and etc., and 14 ESTs remained unknown or hypothetical proteins, which may be new genes. The experiment provided an important basis for studying the disease-resistance mechanism and obtaining the genes for the aim of improving grapevine powdery mildew resistance.展开更多
根据从本课题组前期构建的中国野生葡萄华东葡萄株系白河-35-1在白粉病诱导下的cDNA文库中获得的1条与抗白粉病相关的EST序列,设计特异引物,结合RACE(Rapid-amplrfication of cDNA ends)技术克隆了中国野生华东葡萄白河-35-1广泛逆境蛋...根据从本课题组前期构建的中国野生葡萄华东葡萄株系白河-35-1在白粉病诱导下的cDNA文库中获得的1条与抗白粉病相关的EST序列,设计特异引物,结合RACE(Rapid-amplrfication of cDNA ends)技术克隆了中国野生华东葡萄白河-35-1广泛逆境蛋白基因(Universal stress protein,USP)全长序列。该基因全长836 bp,编码175个氨基酸,GenBank登录号为GU946975。进一步研究该基因在白粉菌诱导下的相对表达水平,我们通过半定量RT-PCR技术,研究了VpUSP在中国野生华东葡萄抗病、感病株系中的表达变化,结果表明,广泛逆境蛋白在中国野生华东葡萄感病株系中,接种白粉病前后均有表达,但表达量变化不明显;然而,抗病的中国野生华东葡萄株系在接种白粉病后VpUSP基因表达量呈明显的增加趋势,初步表明中国野生华东葡萄VpUSP基因在与白粉病菌互作过程中具有表达活性。展开更多
基金funded by the National Natural Science Foundation of China(30571280, 30771493)the Transcentury Talent-Training Program of Ministry of Education of China and the National 863 Project of China(2007AA10Z182)
文摘Uncinula necator is a worldwide serious fungus disease causing annual heavy lost on grapevine production. In order to get more informations on defense related EST sequences and help breeding program, we constructed and characterized a suppression subtractive hybridization cDNA library with artificially inoculated leaves and control Chinese wild Vitis pseudoreticulata clone Baihe-35-1, which is highly resistant to powdery mildew. In the library, the length of 58 EST fragments known as putative functions varied from 130 to 800 bp, and 60% of the ESTs exhibited high similarity to known sequences in database of GenBank with BLASTX analysis. These genes were involved in stress/defense response, detoxification, signal transduction, disease defense, and etc., and 14 ESTs remained unknown or hypothetical proteins, which may be new genes. The experiment provided an important basis for studying the disease-resistance mechanism and obtaining the genes for the aim of improving grapevine powdery mildew resistance.
