中国野生葡萄抗黑痘病基因RAPD标记的克隆、序列分析及辅助育种应用

Cloning and sequence analysis of the RAPD marker linked to anthrac-nose-resistance gene in Chinese wild Vitis and its application for marker-assisted breeding

对中国野生葡萄抗黑痘病基因RAPD标记OPS03-1300进行了克隆、测序及序列分析,结果表明该标记实际长度是1354bp,因此更名为OPS03-1354,其GenBank登录号为DQ350885。该标记序列与99条来自欧洲葡萄的EST有同源性,其中与1条来自赤霞珠叶片感染葡萄皮尔斯病病原菌后获得的EST同源性为82%,与2条来自赤霞珠果实在不同发育期获得的EST同源性分别为80%和85%。该序列与1条来自中国野生华东葡萄白河-35-1叶片接种霜霉病病原菌后获得的EST同源性为67%。该序列编码葡萄的一种假定蛋白。此外,应用该标记对中国野生葡萄与欧洲葡萄的种间杂交广西-1×京可晶F1代339株、白河-35-1×佳利酿F2代207株进行了辅助选择。

Cloning, sequencing and sequence analysis of the RAPD marker OPS03-1300 linked to anthracnose （Elsinoe ampelina）resistant gene in Chinese wild Vitis were carried out. The result showed that the length of the RAPD marker OPS03-1300 was actually 135 4 bp,so it was renamed as OPS03-1354. Nucleotide sequence data of the RAPD marker had been submitted to the GenBank databases ,and the accession number was DQ350885. OPS03-1354 sequence shows identity with 99 EST sequences from Vitis vinifera L.The sequence showed 82% identity with one EST sequence from Cabernet Sauvignon leaves infected with Xylellafastidiosa, 80% and 85% identity respectively with two EST sequences from Cabernet Sauvignon berry of different growth stages. Moreover, the sequence showed 67% identity with one EST sequence from Chinese wild Vitis, Vitis paeudoretlculata clone Baihe-35-1 leaves infected with Plazrnopara vitlcola. The sequence can code a hypothetical protein of grape. In addition, Marker-assisted selection of anthracnose-resistance was performed using OPS03-1354 in hybrids from interspeeies cross between Chinese wild Vitis and Vitis vinifera,including 339 F1 progenies of combination Guangxi-1×Jingkejing and 207 F2 progenies of combination Baihe-35-1×Carignane.