普通菜豆SSR分子标记鉴定体系的建立及应用

Establishment and Application of SSR Molecular Marker Identification System for Common Bean(Phaseolus vulgaris L.)

为了保护育种家合法权益、规范种业市场、提高种子市场监管效率等,本研究拟建立一套普通菜豆品种SSR分子标记鉴定体系。首先,遴选12个产地不同、植物学形态差异较大的普通菜豆品种,通过聚丙烯酰胺凝胶电泳筛选出120对多态性高、条带清晰的SSR引物;然后,利用荧光毛细管电泳技术对筛选出的引物多态性在96个普通菜豆品种中进一步检测,最终获得在菜豆基因组上均匀分布、多态性高、峰型稳定的33对SSR核心引物。在此基础上,依据扩增片段长度和荧光标记颜色对核心引物进行分组,并设置参照品种以校正不同实验批次和平台之间的误差,建立了基于SSR标记的品种鉴定体系。同时,构建了包含申请品种、保护品种、主栽品种和地方品系在内的161个普通菜豆品种的指纹数据库。这套核心引物能够将其中的140个品种区分开,遗传相似系数平均值为0.33,表明这些品种遗传多样性较丰富;未区分开的21个品种在产地、父母本、植物学性状等方面极其相似,但能通过部分性状加以区分,表明该套核心引物在辅助近似品种筛选方面非常有效。本研究建立的基于SSR分子标记的普通菜豆品种鉴定体系将为其真实性鉴定及DUS测试近似品种辅助筛选提供有力支撑。

In recent years,the new varieties breeding of common bean has been developing rapidly,and the number of new varieties and applications for Plant Variety Protection(PVP)has been increasing.In order to protect the legitimate rights and interests of breeders,regulate the seed market and improve the efficiency of the seed market supervision,this study intends to establish common bean identification system based on a set of SSR molecular markers.Firstly,12 common bean varieties with different origins and botanical morphology were selected,and 120 SSR markers with high polymorphism and clear bands were screened by polyacrylamide gel electrophoresis.Then,33 SSR core markers with even distribution,high polymorphism and stable peak pattern were obtained in 96 common bean varieties by fluorescence capillary electrophoresis.On this basis,the core markers were grouped according to the length of amplified fragments and the color of fluorescent markers,and reference varieties were set to correct the errors between different experimental batches and platforms.A variety identification system based on SSR markers was thus established.Meanwhile,a fingerprint database was constructed for 161 common beans for which plant variety protection was applied or which are protected varieties or dominant varieties or landraces.140 varieties can be distinguished by the set of core markers,and the average genetic similarity coefficient is 0.33,showing a high genetic diversity of those varieties.21 varieties that are not distinguished share high similarities in terms of origin,parentage and botanical characters,but they could still be recognized by some traits.The result indicates that the set of core markers are effective in aiding screening of similar varieties.The identification system of common bean based on SSR molecular markers established in this study will provide strong support for variety identification and for assisting selection of similar varieties in DUS tests.