黄瓜性别基因连锁的分子标记筛选

SCREENING OF RAPD MARKER LINKED TO CUCUMBER SEX DETERMINATION GENE

利用RAPD(randomamplifiedpolymorphicDNA)技术对黄瓜性别特异基因进行分子标记连锁研究。共选用 30 0个 1 0碱基的随机引物按BSA法对黄瓜性别表型分离群体进行PCR扩增 ,筛选出 5个在全雌和弱雌株基因池 (genepool)中表现多态性的引物。单株检测表明 ,引物B1 1具有全雌特异性 ,在检测的大多数全雌性单株中均可扩增出一条约 1 0 0 0bp的特征带。而在雌雄同株的单株中则未见。因此 ,将该全雌性特异的片段命名为B1 1 1 0 0 0 。该标记的获得为黄瓜性别特异基因的分离和克隆奠定了基础。另外 ,以ACC合酶基因 (CS ACS1 )特异引物检测分离群体单株 ,发现该酶基因存在于所有性型的单株中 。

Molecular markers linked to the sex determination genes of cucumber were screened with RAPD technology. With the method of Bulked Segregant Analyis, 300 randomly selected 10bp primers were used in PCR amplification of the DNA pools from gynoecious and monoecious cucumber, respectively, and polymorphism were found in five primers in three replications. Those polymorphic primers were used to detect the individual samples with different sex phenotypes, a band of 1000bp was amplified specifically in DNA from the gynoeious plants tested. This band was absent in the monoecious plants and marked as B11 1000 . In addition, primers with 20 bases synthesized according to the 1 aminocyclopropane 1 carboxylic acid synthase gene (CS ACS1) were used to amplify the DNA from individual samples and no specific relationship to either gynoecious or monoecious plants was found.