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园艺学报 ›› 2024, Vol. 51 ›› Issue (2): 219-228.doi: 10.16420/j.issn.0513-353x.2023-0659

• 遗传育种·种质资源·分子生物学 •    下一篇

番茄抗坏血酸含量相关QTL定位及候选基因鉴定

刘根忠1,2,李方曼2,葛平飞2,陶金宝2,张星雨2,叶志彪2,3,张余洋2,3,4,5,*   

  1. 1山东农业大学园艺科学与工程学院,山东泰安 271018;2华中农业大学,果蔬园艺作物种质创新与利用全国重点实验室,武汉 430070;3湖北洪山实验室,武汉 430070;4华中农业大学深圳营养与健康研究院,武汉 430070;5中国农业科学院深圳农业基因组研究所,岭南现代农业科学与技术广东省实验室深圳分中心,广东深圳 518000
  • 出版日期:2024-02-25 发布日期:2024-02-26
  • 基金资助:
    国家重点研发计划项目(2022YFD1200502,2021YFD1200201);国家自然科学基金项目(32372696,31991182);湖北洪山实验室项目(2021hszd007);武汉市生物育种重大专项(2022021302024852);湖北省重点研发计划项目(2022BBA0062,2022BBA0066);湖北省种业高质量发展资金项目(HBZY2023B004);湖北省现代农业产业技术体系建设专项(2023HBSTX4-06);华中农业大学与中国农业科学院深圳农业基因组所合作资金研发项目(SZYJY2023022)

QTL Mapping and Candidate Gene Identification Related to Ascorbic Acid Content in Tomato

LIU Genzhong1,2,LI Fangman2,GE Pingfei2,TAO Jinbao2,ZHANG Xingyu2,YE Zhibiao2,3,and ZHANG Yuyang2,3,4,5,*   

  1. 1College of Horticultural Science and Engineering,Shandong Agricultural University,Tai’an,Shandong 271018,China;2National Key Laboratory of Germplasm Innovation and Utilization of Fruit and Vegetable Horticultural Crops,Huazhong Agricultural University,Wuhan 430070,China;3Hubei Hongshan Laboratory,Wuhan 430070,China;4Shenzhen Institute of Nutrition and Health,Huazhong Agricultural University,Wuhan 430070,China;5Shenzhen Branch of Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology,Shenzhen Institute of Agricultural Genomics,Chinese Academy of Agricultural Sciences,Shenzhen,Guangdong 518000,China
  • Published:2024-02-25 Online:2024-02-26

摘要: 以番茄果实高抗坏血酸含量材料TS-226和低抗坏血酸含量材料TS-228为试验亲本杂交和自交创制F2遗传分离群体,分别构建高抗坏血酸混池和低抗坏血酸混池,集团分离群体测序(bulked segregant analysis sequencing,BSA-seq)分析鉴定到与番茄果实抗坏血酸相关的主效QTL,进一步将与抗坏血酸相关的候选基因定位于番茄基因组8号染色体58.00 ~ 60.15 Mb区域内。结合∆SNP-index数据,推断SlPPO(Polyphenol oxidase)是参与调控番茄抗坏血酸含量的主效基因。以TS-228为背景材料,通过农杆菌介导方法进行遗传转化,获得SlPPO过表达和沉默株系,相比野生型,过表达株系OE-3和OE-18的红熟果实总抗坏血酸含量分别减少了18.89%和26.56%;沉默株系Ri-9和Ri-16红熟果实总抗坏血酸含量分别增加了37.53%和63.93%。综上所述,SlPPO为参与控制番茄红熟果实抗坏血酸含量的关键基因,对果实抗坏血酸含量起负向调控作用。

关键词: 番茄, 集团分离群体测序, 抗坏血酸, QTL, SlPPO

Abstract: The F2 genetic segregating population was constructed via using high ascorbic acid(AsA)tomato accession TS-226 and low ascorbic acid tomato accession TS-228 as parents. The high-ascorbic acid and low-ascorbic acid pools were constructed respectively. Through bulked segregant analysis sequencing analysis(BSA-seq),the main QTL related to ascorbic acid was found,and the candidate gene related to ascorbic acid was located in the 58.00–60.15 Mb region of tomato chromosome 8. Furthermore,the ∆SNP-index data were analyzed to determine the main candidate gene SlPPO(Polyphenol oxidase)that controls the ascorbic acid content of tomato fruit. Using TS-228 as the background plant,transgenic lines with SlPPO overexpression and silencing were obtained by Agrobacterium-mediated transformation. Compared with the wild type,the total ascorbic acid content in the red ripe fruits of the overexpression lines OE-3 and OE-18 decreased by 18.89% and 26.56%,respectively. The total ascorbic acid content in the red ripe fruits of the silenced lines Ri-9 and Ri-16 increased by 37.53% and 63.93% relative to wild type,respectively. Taken together,SlPPO is the key gene to control the ascorbic acid content of tomato red ripe fruit,and plays a negative role in regulating fruit ascorbic acid content.

Key words: tomato, BSA-seq, ascorbic acid, QTL, SlPPO

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