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    林晨俞, 郭鑫, 王文娟, 等. 番茄SlJAZ7抗病功能的研究及与SlTGA7互作的鉴定[J]. 华南农业大学学报, 2024, 45(0): 1-11. doi: 10.7671/j.issn.1001-411X.202312017
    引用本文: 林晨俞, 郭鑫, 王文娟, 等. 番茄SlJAZ7抗病功能的研究及与SlTGA7互作的鉴定[J]. 华南农业大学学报, 2024, 45(0): 1-11. doi: 10.7671/j.issn.1001-411X.202312017
    shu
    LIN Chenyu, GUO Xin, WANG Wenjuan, et al. Study on disease resistance of SlJAZ7 and its interaction with SlTGA7 in tomato[J]. Journal of South China Agricultural University, 2024, 45(0): 1-11. doi: 10.7671/j.issn.1001-411X.202312017
    Citation: LIN Chenyu, GUO Xin, WANG Wenjuan, et al. Study on disease resistance of SlJAZ7 and its interaction with SlTGA7 in tomato[J]. Journal of South China Agricultural University, 2024, 45(0): 1-11. doi: 10.7671/j.issn.1001-411X.202312017
    shu

    番茄SlJAZ7抗病功能的研究及与SlTGA7互作的鉴定

    Study on disease resistance of SlJAZ7 and its interaction with SlTGA7 in tomato

      摘要
    • 摘要:
      目的 细菌性斑点病是导致番茄Solanum lycopersicum减产的主要因素之一，丁香假单胞菌(Pseudomonas syringae pv. tomato DC3000，Pst DC3000)是细菌性斑点病的致病因子之一。瞬时沉默番茄JAZ7基因导致其对Pst DC3000的敏感性增加，然而，验证JAZ7基因抗细菌性斑点病的直接证据及其作用机理的报道较少。本研究从番茄叶片中克隆得到SlJAZ7基因，创制稳定遗传的过表达SlJAZ7的转基因番茄，分析其抗病功能和机理，在转录水平和蛋白水平研究其与抗病性密切相关的SlTGA7的关系，为有效防治细菌性斑点病提供理论基础。
      方法 本研究通过对照和转基因番茄接种Pst DC3000的表型差异分析，鉴定SlJAZ7基因的抗病功能。使用RT-qPCR分析SlJAZ7和SlTGA7基因在Pst DC3000、茉莉酸(Jasmonic acid，JA)和水杨酸(Salicylic acid，SA)处理下的表达模式和组织特异性。利用烟草瞬时表达的方法研究SIJAZ7和SlTGA7蛋白的亚细胞定位。利用酵母双杂交(Y2H)试验、双分子荧光互补(BiFC)试验和蛋白下拉(Pull-down)试验研究SlJAZ7蛋白与SlTGA7蛋白的互作关系，验证SlJAZ7的抗病功能和可能的抗病机理。
      结果 过表达SlJAZ7基因的转基因番茄叶片在Pst DC3000处理时受到的过氧化损伤较野生型更少，转基因株系中SlTGA7表达量升高，SlJAZ7基因在营养组织中高表达，且受到Pst DC3000诱导，响应JA、SA处理，SlTGA7基因在同样的处理下呈相反的变化趋势。SlJAZ7和SlTGA7蛋白均定位于细胞核。Y2H、BiFC和pull-down试验同时证明SlJAZ7蛋白和SlTGA7蛋白存在互作关系。
      结论 过表达SlJAZ7基因有利于减少活性氧积累，提高番茄抗病性，同时SlJAZ7在转录水平正调控SlTGA7基因表达。SlJAZ7与SlTGA7存在互作，推测SIJAZ7基因可能通过提高Pst DC3000病原菌侵染时SITGA7基因的表达量，启动SITGA7下游抗病基因的表达来提高转基因番茄的抗病性，也可能通过与SITGA7蛋白结合，影响其调控MYC等转录因子的活性。这为进一步研究SlJAZ7的作用机制奠定了基础。

       

      Abstract:
      Objective Bacterial spot disease is one of the main factors leading to tomato (Solanum lycopersicum) yield reduction, and Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) is one of the pathogenic factors of bacterial spot disease. Transient silencing of JAZ7 gene in tomato results in increased susceptibility to Pst DC3000. At present, there are a few reports to verify the resistance function and mechanism of JAZ7 gene in tomato. In this study, we cloned SlJAZ7 gene, and a stable genetic transgenic tomato with overexpression of SlJAZ7 was created to analyze its resistance function and mechanism, and the relationship between SlJAZ7 and SlTGA7, which is closely related to disease resistance, was studied at the transcriptional and protein levels, providing a theoretical basis for effective prevention and control of bacterial spot disease.
      Method In this study, the resistance function of SlJAZ7 gene was determined by the phenotypic difference between control and transgenic tomato inoculated with Pst DC3000. RT-qPCR was used to analyze the tissue-specific expression of SlJAZ7 gene and SlTGA7gene under Pst DC3000, JA (Jasmonic acid) and salicylic acid (SA) treatments. The subcellular localization of SIJAZ7 and SlTGA7 proteins was studied by transient expression of tobacco. The interaction between SlJAZ7 and SlTGA7 was verified by Y2H, BiFC and pull-down experiments, to verify the disease resistance function and possible mechanism of SlJAZ7.
      Result Transgenic tomato leaves overexpressing SlJAZ7 gene had lower peroxidation damage than wild type when treated with pathogens, the expression level of SlTGA7 in transgenic lines increased. SlJAZ7 gene was highly expressed in nutrient tissues. SlJAZ7 gene was induced by Pst DC3000 in response to JA and SA treatments. SlTGA7 gene showed opposite change trend under the same treatment. Both SlJAZ7 and SlTGA7 proteins were located in the nucleus. All Y2H, BiFC and pull-down experiments proved the interaction between SlJAZ7 and SlTGA7.
      Conclusion In summary, overexpression of SlJAZ7 gene can reduce the accumulation of reactive oxygen species and improve tomato disease resistance. SlJAZ7 positively regulates SlTGA7 gene expression at the transcriptional level. SlJAZ7 interacts with SlTGA7. It is speculated that SIJAZ7 gene may improve the disease resistance of transgenic tomato by increasing the expression level of SITGA7 gene during pathogen infection, initiating the expression of downstream resistance genes of SITGA7, or by binding with SITGA7 protein, affecting its regulation of MYC and other transcription factors, which lays a foundation for further research on the mechanism of SlJAZ7.

       

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