利用重测序技术开发高粱多态性SSR分子标记

doi:10.13560/j.cnki.biotech.bull.1985.2019-0692

生物技术通报 ›› 2019, Vol. 35 ›› Issue (11): 217-223.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0692

利用重测序技术开发高粱多态性SSR分子标记

王平^{1, 2}, 王春语¹, 张丽霞¹, 丛玲¹, 朱振兴¹, 陆晓春¹

1. 辽宁省农业科学院高粱研究所,沈阳 110161;
2. 沈阳农业大学农学院,沈阳 110161

收稿日期:2019-08-02 出版日期:2019-11-26 发布日期:2019-11-19
作者简介:王平,男,博士,助理研究员,研究方向：高粱分子育种技术研发;E-mail：pingw-556@163.com
基金资助:
国家重点研发计划（2018YFD1000701,2018YFD1000700）,国家现代农业产业技术体系项目（CARS-06-13.5-A3）,国家自然科学基金项目（31301393）

Development of SSR Molecular Markers with Sorghum Polymorphism Using Re-sequencing

WANG Ping^{1, 2}, WANG Chun-yu¹, ZHANG Li-xia¹, CONG Ling¹, ZHU Zhen-xing¹, LU Xiao-chun¹

1. Sorghum Research Institute,Liaoning Academy of Agricultural Sciences,Shenyang 110161;
2. Agronomy College,Shenyang Agricultural University,Shenyang 110161

Received:2019-08-02 Published:2019-11-26 Online:2019-11-19

摘要/Abstract

摘要： SSR分子标记由于具有成本低廉、容易操作等特性,使其在分子标记辅助育种中广泛应用。目前高粱SSR标记多基于测序已经完成美国高粱品种BTX623基因组开发,在应用中筛选多态性标记的效率低。对不育系和恢复系组成的26个高粱材料进行了重测序,然后进行生物信息学分析,最终开发了在26份材料中至少含有2种多态型的SSR标记24 441个。这些SSR标记在26份材料中表现出的基因型多态类型在2-7种之间,2种的有16 694个,7种的仅有77个。另外本研究还进行了筛选单拷贝基因处的多态性SSR分析,共筛选到6 733个。随机挑选均匀覆盖10条染色体的单拷贝基因处SSR标记50对,利用辽宁高粱杂交种辽糯3号进行测试,其中49对能扩增出产物,成功率高达98%。其后利用2个品种和74份微核心种质资源测试表明,50对SSR标记在2个品种中有18对表现出多态性,挑选了一对引物在74份微核心种质中可见8种多态型。本研究表明利用不育系和恢复系材料进行重测序能有效开发多态性高的SSR标记。

关键词: 高粱, SSR, 重测序, 多态性引物, 分子标记

Abstract: SSR marker is widely applied in molecule-assisted breeding due to its low cost and easy operation. Currently,sorghum SSR marker is based on sequencing,and although the genome sequence of American sorghum BTX623 is completed,the efficiency is low in screening polymorphism markers in applications. In this study,26 sorghum materials composed of sterile lines and restorer lines were re-sequenced and analyzed by bioinformatics,and 24 441 SSR markers containing at least 2 polymorphic types in 26 materials were developed. These SSR markers generated 2-7 genotypes in 26 sorghum varieties,including 16 694 markers for 2 genotypes and only 77 markers for 7 genotypes. In addition,6 673 polymorphic SSRs at the single-copy gene were screened. Fifty pairs of SSRs on the 10 evenly-covered sorghum chromosomes were randomly selected as bridge markers,and Liaoning sorghum hybrid Liaonuo 3 was used for test. Among them,49 pairs of primers were amplified,the success rate of which achieved 98%. Subsequent test with 2 sorghum varieties and 74 mini-core germplasm showed that 18 pairs of SSR markers among 50 pairs of SSRs presented polymorphism between the 2 varieties. One selected SSR marker generated 8 genotypes in 74 mini-core germplasm. This study demonstrates that re-sequencing sterile lines and restorer lines may effectively develop SSR markers for Chinese sorghum breeding.

Key words: sorghum, SSR, re-sequencing, polymorphic primer, molecular marker

王平, 王春语, 张丽霞, 丛玲, 朱振兴, 陆晓春. 利用重测序技术开发高粱多态性SSR分子标记[J]. 生物技术通报, 2019, 35(11): 217-223.

WANG Ping, WANG Chun-yu, ZHANG Li-xia, CONG Ling, ZHU Zhen-xing, LU Xiao-chun. Development of SSR Molecular Markers with Sorghum Polymorphism Using Re-sequencing[J]. Biotechnology Bulletin, 2019, 35(11): 217-223.

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利用重测序技术开发高粱多态性SSR分子标记

Development of SSR Molecular Markers with Sorghum Polymorphism Using Re-sequencing

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