普通菜豆SSR分子标记鉴定体系的建立及应用
作者:
作者单位:

1.湖南农业大学园艺园林学院;2.中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程;3.农业农村部科技发展中心

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基金项目:

农业行业标准制订和修订;国家自然科学基金(31772325);湖南省自然科学基金(2017JJ2118)


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

1.College of Horticulture and Landscape, Hunan Agricultural University;2.Institute of Crop Science, Chinese Academy of Agricultural Sciences/ National Key Facility for Crop Gene Resources and Genetic Improvement;3.Development Center of Science and Technology, Ministry of Agriculture and Rural Af airs

Fund Project:

the Establishment and Revision of Agricultural Industry Standards;the National Natural Science Foundation of China ;the Hunan Natural Science Foundation

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    摘要:

    近年来,普通菜豆新品种选育工作快速发展,新选育的品种和品种权申请数量不断增加。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 个多态性高、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 个 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。

    Abstract:

    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.

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陈琼,王兰芬,唐浩,等.普通菜豆SSR分子标记鉴定体系的建立及应用[J].植物遗传资源学报,2019,20(6):1494-1505.

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  • 收稿日期:2019-07-02
  • 最后修改日期:2019-09-24
  • 录用日期:2019-08-20
  • 在线发布日期: 2019-11-19
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