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园艺学报 ›› 2021, Vol. 48 ›› Issue (7): 1329-1339.doi: 10.16420/j.issn.0513-353x.2020-1032

• 研究论文 • 上一篇    下一篇

番茄不规则裂果性状的QTL定位及候选基因分析

陈斌, 吴震, 文军琴, 林昊维, 于璐, 薛灵姿, 周蓉, 蒋芳玲*()   

  1. 南京农业大学园艺学院,农业农村部华东地区园艺作物生物学与种质创新重点实验室,南京 210095
  • 收稿日期:2021-02-07 修回日期:2021-05-25 出版日期:2021-07-25 发布日期:2021-08-10
  • 通讯作者: 蒋芳玲 E-mail:jfl@njau.edu.cn
  • 基金资助:
    国家重点研发计划项目(2019YFD100190200);国家重点研发计划项目(2016YFD0100204);国家自然科学基金项目(32072581);江苏省农业科技自主创新资金项目(CX(20)3101);中央高校基本科研业务费项目(KYZZ201909)

QTL Mapping and Candidate Genes Analysis of Irregular Fruit Cracking in Tomato

CHEN Bin, WU Zhen, WEN Junqin, LIN Haowei, YU Lu, XUE Lingzi, ZHOU Rong, JIANG Fangling*()   

  1. College of Horticulture,Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China,Ministry of Agriculture and Rural Affairs,Nanjing Agricultural University,Nanjing 210095,China
  • Received:2021-02-07 Revised:2021-05-25 Online:2021-07-25 Published:2021-08-10
  • Contact: JIANG Fangling E-mail:jfl@njau.edu.cn

摘要:

为挖掘调控番茄不规则裂果性状的关键基因,利用番茄易发生不规则裂果的种质‘S189’和耐裂果的种质‘R91’杂交获得F1代,之后F1自交构建F2群体。以F2群体中挑选出的20株易裂果单株和20株耐裂果单株的DNA分别等量混合,构成易裂池和耐裂池,进行QTL-seq分析。共检测到2个调控番茄不规则裂果性状的QTL,分别位于2号染色体的38.75 ~ 42.14 Mb区域和5号染色体的49.07 ~ 49.48 Mb区域,暂命名为qCR2qCR5。在qCR5的区域内仅检测到2个基因,利用2个亲本的红熟期果皮进行基因表达量分析,发现其差异均不显著。qCR2的区域较大,为快速有效地挖掘候选基因,利用43个多态性标记构建2号染色体标记连锁图,并结合F2群体裂果率进行遗传连锁分析,在多态性标记sli2734和Bin3371之间检测到1个主效QTL,其LOD值为3.05,贡献率为7.05%,且其对应的物理位置与qCR2的区域相重合。联合QTL-seq分析和遗传连锁分析将候选区域缩小至39.55 ~ 39.94 Mb,记为qCR2.1。在qCR2.1的区域内共有53个基因,利用亲本红熟期果皮进行基因表达量分析,获得3个表达量差异显著的基因Solyc02g072400、Solyc02g072470和Solyc02g076780。其中Solyc02g076780表达量差异极显著,其与乙烯调控相关,推测其为控制番茄不规则裂果性状的关键候选基因。

关键词: 番茄, 不规则裂果, QTL定位, 候选基因

Abstract:

In order to excavate key regulatory genes for irregular fruit cracking in tomato,the F1 and F2 offspring were obtained by crossing the cracking-susceptible(CS)germplasm accession‘S189’and the cracking-resistant(CR)germplasm accession‘R91’and subsequent self-crossing. QTL-seq approach was applied to identify candidate QTLs. Genomic DNA of 20 CS and 20 CR F2 individuals were evenly mixed as CS-pool and CR-pool,respectively. A total of two QTLs that regulated irregular fruit cracking in tomato were detected,which were physically located in the region of 38.75-42.14 Mb on chromosome 2 and the region of 49.07-49.48 Mb on chromosome 5. The two QTLs were temporarily designated as qCR2 and qCR5. In view of the fact that there were only two genes in the region of qCR5,the differences at gene expression levels were analyzed using the mature fruit pericarps from the parents. However,both the results showed no significant differences between the parents. Considering that the region of qCR2 was relatively large,in order to quickly and effectively mine candidate genes,the linkage map of chromosome 2 was constructed using 43 polymorphic markers. Afterwards,traditional QTL analysis was carried out based on the fruit cracking rate of F2 population. A major QTL was detected between the polymorphic markers sli2734 and Bin3371,whose LOD score was 3.05 and contribution rate was 7.05%. The corresponding physical location of the major QTL was coincided with the qCR2’s region. According to the results from both QTL-seq and traditional QTL analysis,the candidate region was delimited between 39.55-39.94 Mb,eventually designated as qCR2.1. There were a total of 53 genes in qCR2.1’s region. Accordingly,the mature fruit pericarps from the parents were used to analyze the differences at gene expression levels and thus three genes with significantly different expression levels were obtained,namely Solyc02g072400,Solyc02g072470 and Solyc02g076780. Among them,the expression levels of Solyc02g076780 between the parents were extremely significantly different. Solyc02g076780 played a role in ethylene regulation,which could be a key candidate gene for irregular fruit cracking in tomato.

Key words: tomato, irregular fruit cracking, QTL mapping, candidate gene

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