Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (4): 1195-1204.DOI: 10.1016/j.jia.2023.08.008

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不结球白菜隐性核不育基因Bra2Ms的精细定位与克隆

  


  • 收稿日期:2023-04-28 接受日期:2023-07-05 出版日期:2024-04-20 发布日期:2024-03-30

Fine mapping and cloning of the sterility gene Bra2Ms in non-heading Chinese cabbage (Brassica rapa ssp. chinensis)

Liping Song1, Xia Li2, Liguang Tang1, Chuying Yu1, Bincai Wang1, Changbin Gao1, Yanfeng Xie1, Xueli Zhang1, Junliang Wang1, Chufa Lin1, Aihua Wang1#   

  1. 1 Wuhan Vegetable Research Institute, Wuhan Academy of Agricultural Sciences, Wuhan 430345, China

    2 Biotechnology and Genetic Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming 650000, China

  • Received:2023-04-28 Accepted:2023-07-05 Online:2024-04-20 Published:2024-03-30
  • About author:Liping Song, E-mail: lp19871120@126.com; #Correspondence Aihua Wang, Tel: +86-27-61801260, E-mail: wangaihualt@163.com
  • Supported by:
    We thank the Wuhan Major Project of Key Technologies in Biological Breeding and New Variety Cultivation, China (2022021302024852), the Science and Technology Support Project of Rural Vitalization in Hubei Province, China (2022BBA121), the Key Research and Development Project of Hubei Province, China (2021BBA097), and the Key Research and Development Project of Hubei Province, China (2021BBA102) for the financial support.

摘要: 雄性不育是植物发育过程中重要的生物学过程,也是目前作物杂种优势利用的重要途径。不结球白菜为典型的异花授粉作物,杂种优势明显,然而,其细胞核雄性不育的分子机理仍不明确。矮脚黄A(WS24-3)为新发现的不结球白菜隐性核雄性不育材料,之前的研究将不育基因Bra2MS锁定在白菜A2染色体上,通过细胞学观察发现,产生不育的原因主要是花粉母细胞减数分裂出现异常。本研究通过精细定位将Bra2MS锁定在129K的物理距离内,结合细胞学数据、转录组数据对这129K序列进行ORF分析,初步确定一个编码PHD-finger蛋白转录因子的Bra039753即是WS24-3A的不育基因Bra2Ms。在减数分裂时期,Bra039753在不育系WS24-3A中明显下调表达。Bra2Ms基因的DNA序列在不育材料中有一段369bp的外源插入,且在第一个外显子上,该插入区段与目标基因是共分离的。这些实验数据将为不结球白菜核不育形成的遗传机制研究提供新线索,本研究中获得的分子标记可辅助加快不结球白菜及其他十字花科作物优良不育系的选育。

Abstract: The application of a male-sterile line is an ideal approach for hybrid seed production in non-heading Chinese cabbage (Brassica rapa ssp. chinensis).  However, the molecular mechanisms underlying male sterility in B. rapa are still largely unclear.  We previously obtained the natural male sterile line WS24-3 of non-heading Chinese cabbage and located the male sterile locus, Bra2Ms, on the A2 chromosome.  Cytological observations revealed that the male sterility of WS24-3 resulted from disruption of the meiosis process during pollen formation.  Fine mapping of Bra2Ms delimited the locus within a physical distance of about 129 kb on the A2 chromosome of B. rapa.  The Bra039753 gene encodes a plant homeodomain (PHD)-finger protein and is considered a potential candidate gene for Bra2MsBra039753 was significantly downregulated in sterile line WS24-3 compared to the fertile line at the meiotic anther stage.  Sequence analysis of Bra039753 identified a 369 bp fragment insertion in the first exon in male sterile plants, which led to an amino acid insertion in the Bra039753 protein.  In addition, the 369 bp fragment insertion was found to cosegregate with the male sterility trait.  This study identified a novel locus related to male sterility in non-heading Chinese cabbage, and the molecular marker obtained in this study will be beneficial for the marker-assisted selection of excellent sterile lines in non-heading Chinese cabbage and other Brassica crops.

Key words: non-heading Chinese cabbage , male sterility ,  Bra2Ms ,  fine mapping ,  PHD-finger protein