中国野生葡萄抗白粉病基因RAPD标记的克隆及序列分析

对获得的中国野生葡萄抗白粉病基因RAPD标记OPW02-1756、OPO11-964、OPY13- 661、OPB11- 520、OPW05-766、OPV03-1365和OPJ16-759进行了回收、克隆、测序及序列分析。结果表明，7个RAPD标记的实际长度分别是1756bp、964bp、661bp、520bp、766bp、1365bp和759bp。OPW02-1756序列与3条欧洲葡萄cDNA克隆获得的EST序列有94 ~ 98%的同源性。OPO11- 964序列与欧洲葡萄“赤霞珠”的1条EST序列有93%同源性，与“霞多丽”的2条果实不同发育时期获得的EST序列有90%和87%的同源性，与甜橙在多种病原菌侵染后通过cDNA文库获得的1条EST序列有83%的同源性。OPO11-964最大的一个阅读框架包含444个碱基，编码147个氨基酸，该氨基酸序列和21个其它生物的未知功能蛋白序列有部分相似性。OPY13-661序列与欧洲葡萄16条EST序列有同源性，其中与“赤霞珠”cDNA克隆的EST有较高的同源性, 与“霞多丽”的2条叶片非生物胁迫有关的EST序列同源性均为89%。OPB11-520序列与拟南芥基因组4条未知功能DNA序列有94%同源性。OPW05-766序列与来自于酿酒葡萄的GAG-POL 前提有50%的同源性。OPV03-1365序列与水稻基因组6条序列有81 ~ 88%的同源性，与拟南芥基因组3条序列有84 ~ 91%的同源性。OPJ16-759与欧洲葡萄“霞多丽”叶片14条非生物胁迫EST序列有83 ~ 100%的同源性，与欧洲葡萄“Cabsau”浆果3条水分胁迫的EST序列有94 ~ 100%的同源性。

Cloning and Sequence Analysis of RAPD Markers Linked to the Resistant Powdery Mildew Genes in Chinese Wild Vitis

OPW02-1756, OPO11-964, OPY13-661, OPB11-520, OPW05-766, OPV03-1365 and OPJ16-759, seven RAPD markers linked to the resistant powdery mildew genes in Chinese wild Vitis were reclaimed, cloned, sequenced and analyzed. The results showed that the real length of seven RAPD markers was 1756bp, 964bp, 661bp, 520bp, 766bp, 1365bp and 759bp respectively. Marker OPW02-1756 shows 94% to 98% identity with 3 EST sequences from European grapes. OPO11-964 shows 93 % identity with 1 EST sequence from Cabernet Sauvignon, shows 90% and 87% identity respectively with 2 EST sequences from green grape berries library, shows 83% identity with 1 EST sequence from multiple pathogen-infected sweet orange. OPO11-964 sequence has a biggest open reading frame of 444bp encoding 147 amino acids, has partial identity with 21 protein sequences of unknown functions from other lives. OPY13-661 sequence has identity with 16 EST sequences from European grapes, and shows 89% identity with 2 EST sequences from abiotic stressed leaves of Vitis vinifera var. Chardonnay. OPB11-520 shows 94% identity with 4 genomic sequences of unknown functions of Arabidopsis thaliana. OPW05-766 shows 50% identity with GAG-POL precursor of wine grape. OPV03-1365 shows 81% to 88% identity with 6 rice genomic sequences, 84% to 91% identity with 3 sequences from Arabidopsis thaliana. OPJ16-759 shows 83% to 100% identity with 14 EST sequences from abiotic stressed leaves of Vitis vinifera var. Chardonnay, 94% to 100% identity with 3 EST sequences from water stressed berry of Vitis vinifera var. Cabsau.