柱花草花粉不育基因连锁的SCAR标记研究

doi:10.11733/j.issn.1007-0435.2020.05.036

草地学报 ›› 2020, Vol. 28 ›› Issue (5): 1478-1483.DOI: 10.11733/j.issn.1007-0435.2020.05.036

柱花草花粉不育基因连锁的SCAR标记研究

黄春琼, 张子雄, 王文强, 刘国道

中国热带农业科学院热带作物品种资源研究所/农业农村部华南作物基因资源与种质创制重点实验室, 海南海口 571101

收稿日期:2020-03-27 修回日期:2020-04-29 发布日期:2020-09-19
通讯作者: 王文强, 刘国道
作者简介:黄春琼(1982-),女,云南宣威人,博士,副研究员;主要从事热带草种质资源与育种研究,E-mail:huangchunqiong44@163.com
基金资助:
国家自然科学基金（31402124）；国家自然科学基金国际合作与交流项目（31861143013）；现代产业技术体系牧草岗位科学家经费（CARS-34）资助

Studies of SCAR Markers Linked to Pollen Sterility Gene in Stylosanthes guianensis

HUANG Chun-qiong, ZHANG Zi-xiong, WANG Wen-qiang, LIU Guo-dao

Tropical Crops Genetic Resources Institute/Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China, Ministry of Agriculture and Rural Affairs, Haikou, Hainan Province 571101, China

Received:2020-03-27 Revised:2020-04-29 Published:2020-09-19

摘要/Abstract

摘要： 本研究以圭亚那柱花草‘1979’（Stylosanthes guianensis ‘1979’，母本，花粉不育）和‘热研2号’柱花草（轮回父本）的BC₁F₁代群体为材料，利用简单序列重复区间扩增多态性（Inter-simple Sequence Repeat，ISSR）技术，结合集群分组分析法（Bulked Segregate Analysis，BSA）构建花粉育性基因池。结果表明：利用100条ISSR引物筛选出在不育基因池中有特异条带的引物共11条；经BC₁F₁群体单株验证，有8个标记仅在花粉不育个体中出现，表明其与花粉不育基因连锁；将这8个特异性片段回收、克隆、测序，并分别设计8对特异序列特异扩增区域（Sequenced Charaeterized Amplified Region，SCAR）引物，对BC₁F₁单株进行验证，发现这8条片段仅在不育个体中扩增出目标条带，表明8 ISSR特异片段成功转化为8个SCAR标记。本研究结果为建立柱花草稳定的雄性不育系提供新的理论基础，为柱花草杂交育种奠定基础。

关键词: 柱花草, ISSR标记, SCAR标记, 不育基因

Abstract: In the present study,the pollen fertility and sterility gene pool were constructed using BC₁F₁ individuals which got through continuous cross from S. guianens 1979 (pollen sterility) and S. guianensis ‘Reyan No.2’ (pollen fertility,recurrent parent) using ISSR technique and BAS (Bulked Segregate Analysis) method. The results showed that 11 ISSR (Inter-simple Sequence Repeat) specific markers of sterile gene pool were screened out using 100 ISSR primers. After individuals of population test,only 8 markers could be amplified in the pollen sterility individuals,which showed 8 markers existed certain linkage relationship with pollen sterility gene. Thus,the 8 markers were collected and sequenced to synthesize 8 SCAR pairs of primers respectively to amplify the individuals of the BC₁F₁ progeny,the results showed that the 8 markers could only be amplified in the pollen sterility individuals,indicating the 8 ISSR markers have been successfully converted into 8 SCAR markers which linked to pollen sterility gene. The research results provide the new theoretical basis for screening specific SCAR markers and establish stable male sterile line of S. guianensis, and lay the foundation for S.guianensis cross breeding.

Key words: Stylosanthes guianensis, ISSR markers, SCAR markers, Sterility gene

中图分类号:

Q943.2

黄春琼, 张子雄, 王文强, 刘国道. 柱花草花粉不育基因连锁的SCAR标记研究[J]. 草地学报, 2020, 28(5): 1478-1483.

HUANG Chun-qiong, ZHANG Zi-xiong, WANG Wen-qiang, LIU Guo-dao. Studies of SCAR Markers Linked to Pollen Sterility Gene in Stylosanthes guianensis[J]. Acta Agrestia Sinica, 2020, 28(5): 1478-1483.

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柱花草花粉不育基因连锁的SCAR标记研究

Studies of SCAR Markers Linked to Pollen Sterility Gene in Stylosanthes guianensis

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