大麻雄性基因连锁AFLP分子标记的筛选及鉴定

摘要

为建立一种大麻早期性别筛选及鉴定方法,利用 AFLP 标记技术,筛选了64对EcoRI-NNN/MseI-NNN引物组合,对11个不同大麻品种雌﹑雄植株的混合DNA池进行了性别连锁特异性条带的筛选。结果表明,6对引物组合表现出多态性,其中, EcoRI-ACA/MseI-CTG在雄性DNA池中扩增出1条特异条带,经各品种单株DNA验证,该条带只在雄性单株稳定出现,回收﹑克隆﹑测序后获得1条可用于大麻早期田间性别鉴定的734 bp雄性特异条带。%64 pairs of AFLP primer combinations of 8 pairs of EcoRI-NNN and MseI-NNN prim-ers were used to find the polymorphisms between mixed DNA pools of Cannabis sativa L. varieties. Six pairs of sixty -four primer combinations showed polymorphism between male and female DNA pools. 700 bp specific fragments were amplified in the primer combination of EcoRI-ACA and MseI-CTG in male DNA pool. This marker was testified with individual DNA of 11 varieties, and the fragment could only be amplified in male plants. The male-specific fragment of 734bp was recovered, cloned and se-quenced.