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 ...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.
