中国野生葡萄抗寒基因的RAPD标记及其序列分析

以抗寒性存在差异的83份葡萄种质及3个杂交组合玫瑰香(Vitis vinifera)×黑龙江实生(V.amurensis)、北醇(V.vinifera×V.amurensis)自交、燕山-1(V.yes hanensis)×河岸-3(V.riparia Beaumont)的F1代122株为试材,进行了中国野生葡萄抗寒基因RAPD标记的研究。通过对110个随机引物的筛选,获得了2个与中国野生山葡萄抗寒基因相连锁的RAPD标记S241-670和S238-800。通过克隆、测序,这2个RAPD标记的实际长度分别是717bp和854bp,因此重新命名为S241-717和S238-854,其序列的GenBank登录号分别为GQ338290和GQ338289。经序列分析,S241-717与29条欧洲葡萄全基因组鸟枪序列有77%~97%的同源性,S238-854与6条欧洲葡萄全基因组鸟枪序列有82%~100%的同源性;二者分别编码欧洲葡萄的一种假定蛋白。RAPD标记S241-717和S238-854可用于葡萄抗寒育种的早期分子标记辅助选择。

RAPD Markers Linked to the Hardiness Gene in Chinese Wild Vitis and Their Sequence analysis

RAPD markers linked to the hardiness gene in Chinese wildVitis were studied with 205 plants, including 83 grape germplasms of different hardiness, 122 F1 individuals from cross combinations Muscat Hamburg （Vitis vinifera ）×Heilongjiang seedling （V.amurensis）, Beichun （V. vinifera×V.amurensis ） selfing and Yanshan-1 （V. yeshanensis ）×V. riparia Beaumont. Two RAPD markers, S241-670 and S238-800 linked to the hardiness gene derived from hardyV. amurensis were obtained by Screening of 110 random primers. Furthermore, cloning and sequencing of the RAPD markers were carried out. The length of the RAPD markers was actually 717 bp and 854 bp. Therefore, they were renamed as S241-717 and S238-854. Nucleotide sequence data of the RAPD markers had been submitted to the GenBank databases,and the accession number was GQ338290 and GQ338289 respectively. Through sequences comparison between the RAPD markers and GenBank, S241-717 and S238-854 show identity with 29 and 6 EST sequences from Vitis vinifera L. respectively. Also, they can code a hypothetical protein of grape respectively. RAPD markers S241-717 and S238-854 could be used as the evidence for marker-assisted selection in grape hardiness breeding.