通过AFLP分子标记方法,应用64对引物组合EcoRⅠ—NNN/MseⅠ—NNN对山葡萄雌花、雄花和两性花植株基因池进行筛选,引物E—AGC/M—CTG在雄花和两性花植株基因池DNA中扩增出一条分子量为283bp的特异性条带;经组池个体和其它品系验证,该特异性条带能在雄株和完全植株中稳定出现。将该特异性条带回收、克隆、测序,设计SCAR标记引物,对已知性别的85份山葡萄种质进行盲测,准确率达到97.6%,表明标记转化成功。由于该SCAR标记在绝大多数山葡萄栽培品种和有育种价值的种质资源中得到验证,可利用该SCAR标记对山葡萄杂交后代在苗期进行性别鉴定,提高育种效率。
Due to its resistant characters to cold, insect and disease, Vitis amurensis Rupr. is one of the most important germplasm resource in grape breeding. It has male, female and hermaphrodite phenotype, but only hermaphrodite is suitable for commercial production. AFLP (amplified fragment length polymorphism ) was applied to identify markers associated with Vitis amurensis Rupr. sex type. By application of 64 markers in the screening, E-AGC/M-CTG was found to amplify a specific 283 bp fragment in male and hermaphrodite pool. According to its sequencing result, SCAR (sequence characterized amplified region) primers were designed, and the primers gave the expected band in 97.6% of the tested 85 cultivars of Vitis amurensis Rupr. It suggested that the SCAR marker will be useful in the seedling stage marker-assistant selection for breeding with the resistant Vitis amurensis Rupr.