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园艺学报 ›› 2025, Vol. 52 ›› Issue (10): 2597-2612.doi: 10.16420/j.issn.0513-353x.2024-0850

• 遗传育种·种质资源·分子生物学 • 上一篇    下一篇

基于重测序的番木瓜基因组InDel标记开发与应用

杨敏*, 贺涵*, 周陈平, 邝瑞彬, 刘传和, 吴夏明, 徐泽, 魏岳荣**()   

  1. 广东省农业科学院果树研究所,农业农村部南亚热带果树生物学与遗传资源利用重点实验室,广东省果树科学与技术研究重点实验室,广州 510640
  • 收稿日期:2025-03-02 修回日期:2025-07-28 出版日期:2025-10-25 发布日期:2025-10-28
  • 通讯作者:
    ** E-mail:
  • 作者简介:

    *共同第一作者

  • 基金资助:
    广州市重点研发计划项目(2023B03J1369); 广东省自然科学基金面上项目(2022A1515010697); 广东农业科学院“中青年学术带头人——金颖之星”培养基金项目(R2023PY-JX005)

Development and Application of Insertion-Deletion(InDel)Markers in Papaya(Carica papaya)Based on Whole Genome Re-Sequencing Data

YANG Min, HE Han, ZHOU Chenping, KUANG Ruibin, LIU Chuanhe, WU Xiaming, XU Ze, WEI Yuerong**()   

  1. Guangdong Provincial Key Laboratory of Science and Technology Research on Fruit Tree;Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization,Ministry of Agriculture and Rural Affairs;Institute of Fruit Tree Research,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China
  • Received:2025-03-02 Revised:2025-07-28 Published:2025-10-25 Online:2025-10-28

摘要:

利用5份番木瓜种质的重测序数据挖掘InDel(插入/缺失)位点,并基于位点设计高效且具有应用价值的分子标记。经严格筛选后,成功鉴定3 635个易于凝胶电泳检测、长度大于30 bp且呈现高度多态性的InDel位点。挑选360个在基因组上均匀分布的位点设计引物,用6份番木瓜材料初筛后发现355对引物能成功扩增出目标产物,其中271对表现出显著的多态性,占候选引物总数的75.3%。从中选取72对高多态性标记对47份番木瓜种质进行基因分型和遗传多样性分析,共检测到145个等位位点;标记的多态性信息含量(PIC)分布于0.0408 ~ 0.4523,平均值为0.3041;基因多样性指数的变异范围为0.1030 ~ 0.8735,平均值为0.5660。通过聚类分析,将47份番木瓜划分为4个类群。此外,基于20对高多态性核心InDel标记的扩增结果,结合花色、单果质量、果形和果肉颜色4种关键农艺性状,构建番木瓜种质的分子身份证。同时,选取能反映亲本间差异的两个InDel标记,鉴定24份番木瓜F1(GT1-5 ב黄金钟’)杂交后代的真实性,成功确认11个真实杂交种。

关键词: 番木瓜, InDel标记, 遗传多样性, F1代真实性鉴定, 分子身份证

Abstract:

InDel(insertion/deletion)loci were mined using resequencing data of 5 papaya germplasm resources,and efficient and applicable molecular markers were designed based on these loci. After strict screening,3 635 InDel loci that are easy to detect by gel electrophoresis,with a length of more than 30 bp and showing high polymorphism were successfully identified. 360 loci evenly distributed across the genome were selected to design primers. After preliminary screening with 6 papaya materials,it was found that 355 pairs of primers could successfully amplify the target products,among which 271 pairs showed significant polymorphism,accounting for 75.3% of the total number of candidate primers. 72 pairs of highly polymorphic markers were selected for genotyping and genetic diversity analysis of 47 papayagermplasms,and a total of 145 allelic loci were detected. The polymorphism information content (PIC)of the markers ranged from 0.0408 to 0.4523,with an average of 0.3041;the gene diversity index varied from 0.1030 to 0.8735,with an average of 0.5660. Through cluster analysis,the 47 papaya accessions were divided into 4 groups. In addition,based on the amplification results of 20 pairs of highly polymorphic core InDel markers,combined with 4 key agronomic traits(flower color,single fruit weight,fruit shape,and flesh color),molecular identities of different papaya germplasms were constructed. Meanwhile,two InDel markers that can reflect the differences between parents were selected to identify the authenticity of 24 F1 hybrid progenies of GT1-5 בHuangjinzhong’papaya,and 11 true hybrids were successfully confirmed.

Key words: papaya, InDel marker, genetic diversity, verification of F1 generation, molecular ID