栽培金鸡菊EST-SSR分子标记开发与亲缘关系分析

doi:10.11733/j.issn.1007-0435.2022.06.014

草地学报 ›› 2022, Vol. 30 ›› Issue (6): 1430-1440.DOI: 10.11733/j.issn.1007-0435.2022.06.014

• 研究论文 • 上一篇

栽培金鸡菊EST-SSR分子标记开发与亲缘关系分析

孙浩男, 李蓉, 李明阳, 郑妍, 刘志杰, 张存石, 刘冬云, 李鹤

河北农业大学园林与旅游学院, 河北保定 071000

收稿日期:2021-11-09 修回日期:2021-12-23 发布日期:2022-07-05
通讯作者: 刘冬云,E-mail:liudongyun0505@163.com;李鹤,E-mail:liheass@163.com
作者简介:孙浩男(1995-),男,汉族,河北唐山人,硕士研究生,主要从事园林植物遗传育种与栽培研究,E-mail:641775049@qq.com
基金资助:
河北省重点研发计划项目（20326339D）；现代种业科技创新专项（21326348D）资助

Development of EST-SSR Markers based on Transcriptome and Analysis of Genetic Relationship of Cultivated Coreopsis

SUN Hao-nan, LI Rong, LI Ming-yang, ZHENG Yan, LIU Zhi-jie, ZHANG Cun-shi, LIU Dong-yun, LI He

College of Landscape and Travel, Hebei Agricultural University, Baoding, Hebei Province 071000, China

Received:2021-11-09 Revised:2021-12-23 Published:2022-07-05

摘要/Abstract

摘要： 为了开发适用于栽培金鸡菊的EST-SSR分子标记，采用二代高通量测序技术对剑叶金鸡菊‘斯丹泰勒’（Coreopsis lanceolata ‘Sterntaler’）的叶片进行转录组测序。结果表明：通过转录组测序共得到42 958条Unigene，利用MISA软件共检测出个6 546个SSR位点，平均分布距离6.36 kb。除单核苷酸外，重复类型以二核苷酸和三核苷酸为主，分别占总数的19.28%和30.13%，二者优势重复单元和占比分别为AG/CT （57.30%）和AAT/ATT （27.94%）。通过Primer3软件设计得到4 499条引物，在随机筛选的50对引物中成功开发出22对多态性较好的引物，扩增出的166个条带全部具有多态性。通过UPGMA聚类分析，在遗传距离为0.460时可将100个栽培金鸡菊种质划分为7类，其遗传相似系数介于0.542 2~1之间，平均为0.747 6。金鸡菊组、两色组、歧序组和轮叶组相关的种质可作为金鸡菊远缘杂交育种的材料。本研究为栽培金鸡菊的分类鉴定、杂交育种和绘制指纹图谱提供了理论依据。

关键词: 金鸡菊, 种质资源, EST-SSR, 转录组, 亲缘关系

Abstract: To develop EST-SSR molecular markers for cultivated Coreopsis,the transcriptome of the of Coreopsis lanceolata ‘Sterntaler’ was sequenced by using high-throughput sequencing technology. Based on the sequencing data,we selected some polymorphic primers for the genetic relationship of 100 germplasm resources of Coreopsis. The results showed that a total of 42 958 unigenes were obtained and a total of 6 546 SSR locis were detected by using MISA software. Except fot mono-nucleotides,the repeat types were mainly di-nucleotides and tri-nucleotides,accounting for 19.28% and 30.13%,with the dominant repeat units and ratios of AG/CT (57.30%) and AAT/ATT (27.94%),respectively. Total 4 499 primers were designed by Primer 3 software. 22 pairs of EST-SSR primers with good polymorphism were successfully developed from these data. A total of 166 bands were amplified and the percentage of polymorphic bands was 100%. Based on the UPMGA clustering analysis,100 Coreopsis germplasms could be divided into 7 groups at a genetic distance of 0.664. The genetic similarity coefficients ranged from 0.542 2 to 1,with an average of 0.747 6. Germplasms related to C. sect. Coreopsis,C. sect. Calliopsis,C. sect. Eublepharis and C. sect. Gyrophyllum can be used as materials for distant hybrid breeding in Coreopsis. This study provides a theoretical basis for the classification,identification,breeding and genetic map construction of cultivated Coreopsis.

Key words: Coreopsis, Germplasm resource, EST-SSR, Transcriptome, Genetic relationship

中图分类号:

孙浩男, 李蓉, 李明阳, 郑妍, 刘志杰, 张存石, 刘冬云, 李鹤. 栽培金鸡菊EST-SSR分子标记开发与亲缘关系分析[J]. 草地学报, 2022, 30(6): 1430-1440.

SUN Hao-nan, LI Rong, LI Ming-yang, ZHENG Yan, LIU Zhi-jie, ZHANG Cun-shi, LIU Dong-yun, LI He. Development of EST-SSR Markers based on Transcriptome and Analysis of Genetic Relationship of Cultivated Coreopsis[J]. Acta Agrestia Sinica, 2022, 30(6): 1430-1440.

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栽培金鸡菊EST-SSR分子标记开发与亲缘关系分析

Development of EST-SSR Markers based on Transcriptome and Analysis of Genetic Relationship of Cultivated Coreopsis

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