洋葱雄性不育恢复位点Ms座位的SSR标记开发及其应用

doi:10.11924/j.issn.1000-6850.casb18120064

中国农学通报 ›› 2020, Vol. 36 ›› Issue (11): 49-55.doi: 10.11924/j.issn.1000-6850.casb18120064

所属专题：园艺

洋葱雄性不育恢复位点Ms座位的SSR标记开发及其应用

杨亚会¹^,², 刘冰江², 杨妍妍², 吴雄², 曹辰兴¹(), 霍雨猛²()

¹山东农业大学园艺科学与工程学院,山东泰安 271018
²山东省农业科学院蔬菜花卉研究所/山东省设施蔬菜生物学重点实验室/国家蔬菜改良中心山东分中心,济南 250100

收稿日期:2018-12-17 修回日期:2019-02-07 出版日期:2020-04-15 发布日期:2020-04-28
通讯作者: 曹辰兴,霍雨猛
作者简介:杨亚会,女,1993年出生,山东菏泽人,硕士研究生,研究方向:蔬菜遗传育种。通信地址:250100 山东省济南市历城区山东省农业科学院蔬菜花卉研究所,Tel:0531-66659015,E-mail: m15098337277@163.com
基金资助:
国家自然科学基金“洋葱雄性不育位点完全连锁基因AcSKP1的功能及互作蛋白的挖掘”(31672165);“洋葱果胶甲酯酶基因AcPME的功能研究”(31201635);山东省农业科学院青年英才培养计划(IVFSAAS2016-2018-01)

The Development and Application of SSR Markers Linked to Male Sterility Restoration Site Ms Locus in Allium cepa L.

Yang Yahui¹^,², Liu Bingjiang², Yang Yanyan², Wu Xiong², Cao Chenxing¹(), Huo Yumeng²()

¹College of Horticulture Science and Engineering, Shandong Agricultural University, Taian Shandong 271018
²Institute of Vegetables and Flowers, Shandong Academy of Agricultural Sciences/Shandong Provincial Key Laboratory for Biology of Greenhouse Vegetable/Shandong Branch of National Improvement Center for Vegetable, Jinan 250100

Received:2018-12-17 Revised:2019-02-07 Online:2020-04-15 Published:2020-04-28
Contact: Cao Chenxing,Huo Yumeng

摘要/Abstract

摘要：

本研究旨在开发稳定可靠的洋葱育性位点分子标记,并将其应用于育种实践,筛选育种系,从而缩短育种周期,节省育种成本,加速洋葱杂交种的培育进程。以洋葱育性恢复Ms座位的AFLP序列为基础,采用序列比对分析、PCR检测和表型分析等方法,开发、鉴定、应用了WH-SSR-1分子标记。结果表明:WH-SSR-1标记与Ms位点紧密连锁,Ms位点基因型为纯合显性MsMs时,有1条102 bp的扩增带;纯合隐性msms时,有1条99 bp的扩增带;杂合Msms时,有102 bp和99 bp两条扩增带。32份洋葱材料的验证结果证实了Ms座位的标记类型与基因型完全相符。随后从4个OP群体中直接获得了2个配套的不育系与保持系,‘吊玉’和‘天正红玉’因未检测到保持株或不育株,无法简单实现配套。WH-SSR-1标记与Ms位点紧密连锁,能够将其用于育种系筛选的育种实践,从OP群体中直接选择保持株和不育株,同时实现两系配套。

关键词: 洋葱, 保持系, 雄性不育系, Ms位点, SSR标记

Abstract:

This study aims to develop a stable and reliable molecular marker of onion fertility locus and apply it to breeding practice for screening breeding lines, which will further shorten the breeding cycle, save breeding costs, and accelerate the cultivation process of onion hybrids. Based on the AFLP sequences of onion fertility restoration Ms locus, WH-SSR-1 molecular marker was developed, identified and applied in breeding practice by sequence alignment analysis, PCR detection and phenotype analysis. The results showed that WH-SSR-1 marker was closely linked to Ms locus. When Ms locus genotype was homozygous dominant MsMs, there was one 102 bp band; when Ms locus genotype was homozygous recessive msms, there was only one 99 bp band; when Ms locus genotype was hybridized Msms, there were two bands, 102 bp band and 99 bp band, respectively. The validation results of 32 onion materials confirmed that the marker type of Ms locus was completely consistent with the genotype. Subsequently, two matching maintainer and sterile lines were directly developed from four OP populations, but ‘Diaoyu’ and ‘Tianzhenghongyu’ could not be easily matched because no maintainer or sterile individuals were detected. The WH-SSR-1 marker is closely linked to Ms locus and can be used in the breeding practice. The maintainer and sterile individuals can be directly selected from OP population, and can be matched with each other and form counterpart breeding lines at the same time.

Key words: onion (Allium cepa L.), maintainer line, male sterile line, Ms locus, SSR marker

中图分类号:

S633.2

杨亚会, 刘冰江, 杨妍妍, 吴雄, 曹辰兴, 霍雨猛. 洋葱雄性不育恢复位点Ms座位的SSR标记开发及其应用[J]. 中国农学通报, 2020, 36(11): 49-55.

Yang Yahui, Liu Bingjiang, Yang Yanyan, Wu Xiong, Cao Chenxing, Huo Yumeng. The Development and Application of SSR Markers Linked to Male Sterility Restoration Site Ms Locus in Allium cepa L.[J]. Chinese Agricultural Science Bulletin, 2020, 36(11): 49-55.

图/表 6

参考文献 31

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品系或杂交种	来源¹	细胞质²	Ms的基因型³	表型(+/-)⁴	性状⁵
118	SAAS	S	msms	-	EM,黄色
12-10	SAAS	S	MsMs	+	MEM,黄色
110	SAAS	S	msms	-	EM,黄色
502	SAAS	S	msms	-	MEM,黄色
503	SAAS	S	msms	-	MEM,黄色
101	SAAS	S	msms	-	MEM,黄色
117-5	SAAS	S	msms	-	MEM,红色
117-11-1	SAAS	S	msms	-	MEM,红色
117-10-1	SAAS	S	msms	-	MEM,红色
117-10-2	SAAS	S	msms	-	MEM,红色
117-10-3	SAAS	S	msms	-	MEM,红色
210	SAAS	N	msms	+	EM,黄色
218	SAAS	N	msms	+	EM,黄色
602	SAAS	N	msms	+	MEM,黄色
603	SAAS	N	msms	+	MEM,黄色
202	SAAS	N	msms	+	MEM,黄色
217-5	SAAS	N	msms	+	MEM,红色
217-11-1	SAAS	N	msms	+	MEM,红色
217-10-1	SAAS	N	msms	+	MEM,红色
217-10-2	SAAS	N	msms	+	MEM,红色
217-10-3	SAAS	N	msms	+	MEM,红色
324	SAAS	S	MsMs	+	MEM,黄色
327	SAAS	S	MsMs	+	MEM,黄色
231	SAAS	N	MsMs	+	MEM,黄色
234	SAAS	N	MsMs	+	MEM,黄色
TABAO	TakII Seed	S	Msms	+	M,黄色
ATON	TakII Seed	S	Msms	+	MLM,黄色
EARTH	TakII Seed	S	Msms	+	MLM,黄色
ADVANCE	Shippo Seed	S	Msms	+	MEM,黄色
AMA70	Shippo Seed	S	Msms	+	MEM,黄色
TASAN	Shippo Seed	S	Msms	+	M,黄色
MOMIJI No.3	Shippo Seed	S	Msms	+	MLM,黄色

品系或杂交种	来源¹	细胞质²	Ms的基因型³	表型(+/-)⁴	性状⁵
118	SAAS	S	msms	-	EM,黄色
12-10	SAAS	S	MsMs	+	MEM,黄色
110	SAAS	S	msms	-	EM,黄色
502	SAAS	S	msms	-	MEM,黄色
503	SAAS	S	msms	-	MEM,黄色
101	SAAS	S	msms	-	MEM,黄色
117-5	SAAS	S	msms	-	MEM,红色
117-11-1	SAAS	S	msms	-	MEM,红色
117-10-1	SAAS	S	msms	-	MEM,红色
117-10-2	SAAS	S	msms	-	MEM,红色
117-10-3	SAAS	S	msms	-	MEM,红色
210	SAAS	N	msms	+	EM,黄色
218	SAAS	N	msms	+	EM,黄色
602	SAAS	N	msms	+	MEM,黄色
603	SAAS	N	msms	+	MEM,黄色
202	SAAS	N	msms	+	MEM,黄色
217-5	SAAS	N	msms	+	MEM,红色
217-11-1	SAAS	N	msms	+	MEM,红色
217-10-1	SAAS	N	msms	+	MEM,红色
217-10-2	SAAS	N	msms	+	MEM,红色
217-10-3	SAAS	N	msms	+	MEM,红色
324	SAAS	S	MsMs	+	MEM,黄色
327	SAAS	S	MsMs	+	MEM,黄色
231	SAAS	N	MsMs	+	MEM,黄色
234	SAAS	N	MsMs	+	MEM,黄色
TABAO	TakII Seed	S	Msms	+	M,黄色
ATON	TakII Seed	S	Msms	+	MLM,黄色
EARTH	TakII Seed	S	Msms	+	MLM,黄色
ADVANCE	Shippo Seed	S	Msms	+	MEM,黄色
AMA70	Shippo Seed	S	Msms	+	MEM,黄色
TASAN	Shippo Seed	S	Msms	+	M,黄色
MOMIJI No.3	Shippo Seed	S	Msms	+	MLM,黄色

分离群体	可育S (Msms)		不育S (msms)		总个体数	交换值/%
分离群体	二带	一带	二带	一带	总个体数	交换值/%
BC_1-1	61	0	0	54	115	0
BC_1-2	61	0	0	63	124	0

分离群体	可育S (Msms)		不育S (msms)		总个体数	交换值/%
分离群体	二带	一带	二带	一带	总个体数	交换值/%
BC_1-1	61	0	0	54	115	0
BC_1-2	61	0	0	63	124	0

Ops （栽培种）	特性	鳞茎数	WH-SSR-1/accD标记鉴定		子代表型
Ops （栽培种）	特性	鳞茎数	不育株	保持株	不育株子代	保持株子代
吊玉	黄色,中熟	50	13	0	-	-
天正黄金	金黄色,中晚熟	50	9	3	不育	可育
晚生大玉葱	棕黄色,中晚熟	50	4	15	不育	可育
天正红玉	红色,中晚熟	50	0	9	不育	可育

洋葱雄性不育恢复位点Ms座位的SSR标记开发及其应用

The Development and Application of SSR Markers Linked to Male Sterility Restoration Site Ms Locus in Allium cepa L.

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