基于SSR标记的5个西瓜新品种纯度鉴定及特异性分析的研究

Seed Purity Detection and Distinctiveness Analysis of 5 New Watermelon Hybrid Varieties on SSR Markers

为建立西瓜杂交种品种纯度的快速、准确、高效鉴定体系,利用SSR分子标记,对5个西瓜新品种纯度进行鉴定,并构建亲本的分子指纹图谱。以上海市农业科学院育成的5个西瓜新品种‘申选958’、‘双色冰淇淋’、‘黄晶’、‘圣女红2号’与‘圣女红3号’及其亲本的种子为试材,采用CTAB法从发芽种子的根尖中提取DNA。根据杂交种带型双亲互补的原则,利用SSR标记进行杂交种纯度及特异性鉴定研究。从28对SSR核心引物中筛选出5对特异性引物(引物BVWS00106、BVWS00209、BVWS0036、BVWS01734、BVWS00826),鉴定到这5个新品种的杂交种纯度均达到98%,并与各品种的大田植物学形态特征鉴定结果相比较,符合率高达98%以上。分析比较各亲本材料的SSR引物的多态性表现,构建了9份亲本材料的DNA指纹图谱。研究为西瓜杂交品种纯度快速检测和品种权保护提供了理论依据与技术支撑。

In order to establish a rapid, acute and effective seed purity detection system of watermelon hybrids, the variety purity of 5 watermelon hybrids was identified, and the molecular fingerprint of parental germplasms was constructed by using SSR molecular markers. With seeds of 5 new watermelon hybrid varieties ‘ Shenxuan 958＇, ‘Shuangsebingqilin＇, ‘ Huangjing＇,‘Shengnvhong 2＇ and ‘Shengnvhong 3＇ together with their parents which were bred by researchers of Shanghai Academy of Agricultural Sciences as test materials, total DNA was extracted from root tips of germinating seed by CTAB. Based on the principle of complementary with female parent and male parent of SSR electrophoresis band which was a hybrid variety, the variety purity and the distinctiveness of these watermelon hybrids were identified by analysis of SSR molecular markers. Five pairs of polymorphic primers （primer BVWS00106, BVWS00209, BVWS0036, BVWS01734 and BVWS00826） out of 28 pairs of SSR core primers were selected to make rapid variety purity identification, and the seed purity ofthese 5 watermelon hybrids were 98%. Compared with results of morphological identification in field, the coincidence rate of the variety purity was higher than 98%, which was obtained by SSR analysis. And the fingerprints of 9 parental germplasms of the 5 watermelon hybrids were constructed by using polymorphic sites of the selected SSR markers. The results showed that the technical system could meet the requirements of rapid variety puritv detection and protection of plant variety right of watermelon new hybrids in laboratory.