大豆细胞质雄性不育恢复基因 GmRf1 的精细定位
作者:
作者单位:

1.吉林农业大学农学院;2.吉林省农业科学院大豆研究所 / 大豆国家工程研究中心

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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
Author:
Affiliation:

1.College of Agronomy,Jilin Agricultural University;2.Soybean Research Institute,Jilin Academy of Agricultural Sciences/The National Engineering Research Center for Soybean

Fund Project:

Jilin Province Agricultural Science and Technology Innovation Project(CXGC2021ZD002,CXGC2021RCY003),China Agriculture Research System(CARS-04)

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    摘要:

    目前大豆杂交育种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被测恢复系与不育系测交后代 F1 的育性确定,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;利用集群分离 分析法(BSA,bulked segregant analysis)对双亲、可育和半不育池进行测序分析,发现该基因位点位于 16 号染色体上;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。与 ZH13 v2.0 参考基 因组进行比对发现,GmRf1 位于 16 号染色体 32 708 896~32 932 950 bp 之间,物理距离约为 224.1 kb。本研究为今后分子标 记辅助选育含 GmRf1 基因位点的恢复系和克隆 GmRf1 基因奠定了基础。

    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 tested 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 markerassisted 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 both parents and two pools(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. This study will lay the foundation for molecular marker assisted breeding of the restorer lines containing GmRf1 locus and the isolation of the GmRf1 gene.

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引用本文

郭凤兰,林春晶,王鹏年,等.大豆细胞质雄性不育恢复基因 GmRf1 的精细定位[J].植物遗传资源学报,2022,23(2):518-526.

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  • 收稿日期:2021-08-27
  • 最后修改日期:2021-09-10
  • 录用日期:2021-09-24
  • 在线发布日期: 2022-03-10
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