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  • 颜新林,管中荣,温雯,等.基于SSR标记的芥菜品种鉴定技术体系建立及应用[J].植物遗传资源学报,2021,22(3):758-770.    [点击复制]
  • Yan Xin-lin,Guan Zhong-rong,Wen Wen,et al.Establishment and Application of Mustard Variety Identification System Based on SSR Markers (Brassica juncea L.)[J].植物遗传资源学报,2021,22(3):758-770.   [点击复制]
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基于SSR标记的芥菜品种鉴定技术体系建立及应用
颜新林1, 管中荣2, 温雯3, 张浙峰4, 王晨宇3, 沈进娟2, 汪启明1, 韩瑞玺3, 饶力群1
0
(1.湖南农业大学生物科学技术学院;2.重庆市渝东南农业科学院;3.农业农村部科技发展中心;4.四川省农业科学院作物研究所)
摘要:
利用SSR分子标记建立芥菜品种鉴定体系,旨在为我国芥菜育成品种鉴定提供高通量、快速鉴定方法。选取芥菜16个变种代表性品种用于SSR引物初筛,通过聚丙烯酰胺凝胶电泳从432对引物中筛选出84对条带清晰、稳定性好、多态性高的芥菜SSR引物,并对筛选出的引物5’端进行荧光标记。另外选取96份品种对上述84对引物进行复筛,利用基因分析仪对所有扩增片段大小进行分析,依据各引物的扩增稳定性、峰型易读取程度、多态性及染色体位置等,最终筛选出25对SSR引物作为核心引物,并为其设置参照品种以消除不同实验批次或平台等因素引起的误差。根据荧光颜色和扩增片段大小将25对核心引物分成6组,建立基于SSR标记的芥菜品种鉴定体系。利用该套核心引物构建了189份芥菜品种的DNA指纹数据库,共检测到175个等位变异,平均每对引物检测到7个等位变异;基因多样性指数变化为0.239~0.870,平均为0.555;多态性信息含量在0.23~0.86之间,平均为0.493;本研究建立的基于毛细管电泳平台的芥菜SSR分子标记品种鉴定体系,可用于芥菜品种真实性快速鉴定、遗传多样性分析和DUS测试近似品种辅助筛选等。
关键词:  芥菜  SSR标记  品种鉴定  DNA指纹图谱
DOI:10.13430/j.cnki.jpgr.20201014002
投稿时间:2020-10-14修订日期:2020-10-14
基金项目:农业行业标准制订和修订;湖南省发改委科技专项课题《湖南省主要中药材DNA条形码信息采集与应用》(湘发改投资[2014]658号)
Establishment and Application of Mustard Variety Identification System Based on SSR Markers (Brassica juncea L.)
Yan Xin-lin1, Guan Zhong-rong2, Wen Wen3, Zhang Zhe-feng4, Wang Chen-yu3, Shen Jin-juan2, Wang Qi-ming1, Han Rui-xi3, Rao Li-qun1
(1.College of Bioscience and Biotechnology, Hunan Agricultural University;2.Chongqing Yudongnan Academy of Agricultural Science;3.Development Center of Science and Technology, Ministry of Agriculture and Rural Affairs;4.Institute of Crop Science,Sichuan Academy of Agricultural Sciences)
Abstract:
This study attempted to establish a high-throughput and rapid method using SSR markers, in order to clarify the mustard varieties (Brassica juncea L.) released from China. Sixteen representative mustard varieties were used to screen SSR primers showing polymorphism. Out of 432 pairs of tested primers, 84 were detected with higher polymorphism and good repeatability, and they were subjected for synthesizing the fluorescent-labeled primers. We further genotyped 96 varieties with updated versions of primers using gene analyzer. Based on the amplification stability, the readability of peaks, polymorphism and chromosome location, etc., 25 SSR markers were finally qualified. We further provided the solution using reference varieties to eliminate genotyping errors caused by different experimental batches or platforms. By taking use of these markers from six groups according to the fluorescence color and amplification fragment size, a SSR-based mustard variety identification system has been established. The DNA database containing the fingerprinting information of 189 mustard varieties was generated showing a total of 175 allelic variants with an average of 7 allelic variants per primer. The gene diversity index was variable from 0.239 to 0.870, with an average of 0.555, while the polymorphism was observed between 0.23 and 0.86, with an average of 0.493. Collectively, this study established a mustard variety clarification method by taking use of 25 SSR markers on the capillary electrophoresis genotyping platform, which might be useful in rapid authenticity identification, genetic diversity analysis and similar varieties selection in DUS test in mustard.
Key words:  Mustard  SSR marker  Variety identification  DNA fingerprint

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