Citation:[Copy]
[Copy]
【Print page】 【Online reading】【PDF Full text】 查看/发表评论下载PDF阅读器关闭

Back Issue    Advanced search

This Paper:Browse 629   Download 0  
大豆细胞质雄性不育育性恢复基因GmRf1的精细定位
郭凤兰1,2, 林春晶2, 王鹏年2, 杨绪磊1,2, 吴 铮1,2, 彭 宝2, 赵丽梅1,2, 张春宝1,2
0
(1吉林农业大学农学院,长春130118;2吉林省农业科学院大豆研究所/大豆国家工程研究中心,长春130033)
摘要:
目前大豆杂交种选育主要依赖于以细胞质雄性不育(CMS,Cytoplasmic Male Sterility)为基础的“三系”法。这其中恢复系的选育至关重要,恢复基因的有无和恢复能力的强弱主要通过与不育系测交后代F1的育性来鉴定,既耗时又费力。若能定位和克隆恢复基因(Rf,Restorer-of-fertility),用于恢复系的鉴定或辅助选育将大幅提高选育效率。本研究以大豆CMS-RN型不育系JLCMS204A为母本与含有未知Rf基因的恢复系父本JLR230进行杂交获得的F2分离群体为材料,通过花粉育性鉴定,明确了该恢复系所含恢复基因受一对显性单基因控制,符合单基因配子体遗传模式;利用混合群体分离分析法(BSA,Bulked Segregant Analysis)对亲本、可育和半不育池进行测序分析,将该基因位点位于16号染色体;利用简单重复序列(SSR,Simple Sequence Repeat)分子标记进行多态性分析,初步将该基因定位于标记BARCSOYSSR_16_1069和BARCSOYSSR_16_1076之间,命名为GmRf1。利用酶切扩增多态性序列 (dCAPS,Derived Cleaved Amplified Polymorphic Sequences)标记、插入缺失(InDel,Insertion-Deletion)标记和序列标签位点(STS,Sequence Tag Site)标记,进一步精细定位,最终将GmRf1定位在标记dCAPS-1和BARCSOYSSR_16_1076之间,遗传距离分别为0.1 cM和0.3 cM。以ZH13 v2.0参考基因组进行比对发现,GmRf1位于16号染色体32 708 896 bp~32 932 950 bp之间,物理距离为224.1 kb。本研究通过BSA测序结合新开发分子标记,精细定位了恢复基因GmRf1,这为今后分子标记辅助选育含GmRf1基因位点的恢复系提供了技术支撑,也为后续GmRf1基因的克隆奠定了理论基础。
关键词:  大豆  细胞质雄性不育  恢复基因  分子标记
DOI:10.13430/j.cnki.jpgr.20210827002
投稿时间:2021-08-27修订日期:2021-09-10
基金项目:吉林省农业科技创新工程项目(CXGC2021ZD002,CXGC2021RCY003);国家现代农业产业技术体系建设专项(CARS-04)
Fine Mapping of a Restorer-of-fertility Gene GmRf1 for the Cytoplasmic Male Sterility in Soybean
GUO Feng-lan1,2, LIN Chun-jing2, WANG Peng-nian2, YANG Xu-lei1,2, WU Zheng1,2, PENG Bao2, ZHAO Li-mei1,2, ZHANG Chun-bao1,2
(1 College of agronomy, Jilin Agricultural University, Changchun 130118; 2Soybean Research Institute, Jilin Academy of Agricultural Sciences/The National Engineering Research Center for Soybean, Changchun 130033)
Abstract:
The hybrid breeding in soybean (Glycine max) mainly relies on the three-lines system derived from cytoplasmic male sterility. Identification of optimal restorer lines is therefore of importance. Since checking the fertility of F1 hybrids derived from the restorer lines crossing with sterile lines is time-consumption and laborious, genetic identification of the strong restorer-of-fertility (Rf) gene(s) and its use for marker-assisted selection (MAS) will greatly improve the efficiency of breeding. In this study, the F2 segregation population derived from the CMS-RN type sterile line JLCMS204A (female parent) and the restorer line JLR230 (male parent, containing the unknown Rf gene) was investigated. Through examining pollen fertility the Rf gene in the restorer line was controlled by a pair of dominant single gene. Based on the bulked segregant analysis (BSA) of four pools (both parents, fertile and semi-sterile), the genetic locus named GmRf1 was allocated on chromosome 16 flanked by simple sequence repeat (SSR) markers BARCSOYSSR_16_1069 and BARCSOYSSR_16_1076. By taking use of enzyme digestion amplified polymorphic sequences (dCAPS) makers, insertion deletion (InDel) makers and sequence tag site (STS) makers, GmRf1 was finally delimited between the marker dCAPS-1 and BARCSOYSSR_16_1076, in which the genetic distance were 0.1 cM and 0.3 cM, respectively. On the basis of the ZH13 v2.0 reference genome, GmRf1 was located between 32 708 896 bp and 32 932 950 bp with a physical distance of 224.1 kb. Altogether, the fine mapping of the restorer-of-fertility gene GmRf1 was accomplished, which provided a basis for molecular marker assisted breeding of the restorer lines containing GmRf1 locus and the isolation of the GmRf1 gene.
Key words:  Soybean  Cytoplasmic male sterility  Restorer-of-fertility gene  Molecular marker