{"title":"14‐3‐3蛋白SlTFT1通过结合和稳定乙烯信号通路中的YFT1加速番茄果实成熟","authors":"Tengjian Wen, Lichun Cao, Lida Zhang, Lingxia Zhao","doi":"10.1111/pbi.70274","DOIUrl":null,"url":null,"abstract":"Ethylene plays a pivotal role during the fruit ripening process in tomato (<jats:styled-content style=\"fixed-case\"><jats:italic>Solanum lycopersicum</jats:italic></jats:styled-content>). Previously, we have reported that the tomato EIN2‐like protein YELLOW‐FRUITED TOMATO1 (YFT1), a core component in the ethylene signal transduction pathway, exerts a critical regulatory function in the tomato fruit ripening process. However, the molecular mechanism of YFT1‐mediated ethylene signalling during tomato ripening still remains largely unknown. In this study, we performed yeast two‐hybrid screens of a cDNA expression library from tomato fruits and obtained a new YFT1‐interacting protein, SlTFT1 (TOMATO FOURTEEN‐THREE PROTEIN 1), and confirmed their interaction in vitro and in vivo. Further bimolecular fluorescence complementation (BiFC) assays indicated that SlTFT1 can specifically bind the canonical and non‐canonical on the carboxyl terminus of YFT1 (YFT1‐C). Triple response assays and ethylene responsive gene expression analysis demonstrated that SlTFT1 positively regulates the ethylene signalling pathway in a YFT1‐dependent manner. Phenotypical and biochemical analysis of SlTFT1 overexpression lines and loss‐of‐function mutants showed that <jats:italic>SlTFT1</jats:italic> significantly accelerated ethylene emission, chromoplast development, lycopene accumulation and fruit ripening rates; however, the deletion of <jats:italic>YFT1</jats:italic> and/or <jats:italic>SlTFT1</jats:italic> lesion generated the opposite results, suggesting that <jats:italic>SlTFT1</jats:italic> positively regulates tomato fruit ripening also in a YFT1‐dependent manner. Co‐expression and western blotting assays showed that SlTFT1 can efficiently prevent YFT1 from protein degradation mediated by an F‐box protein SlETP2‐like3, suggesting that SlTFT1 can stabilise YFT1 through their physical interaction. Collectively, our results reveal a genetic and molecular framework of the SlTFT1‐YFT1 complex, which modulates tomato fruit ripening by regulating ethylene signalling.","PeriodicalId":221,"journal":{"name":"Plant Biotechnology Journal","volume":"91 1","pages":""},"PeriodicalIF":10.5000,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"The 14‐3‐3 Protein SlTFT1 Accelerates Tomato Fruit Ripening by Binding and Stabilising YFT1 in the Ethylene Signalling Pathway\",\"authors\":\"Tengjian Wen, Lichun Cao, Lida Zhang, Lingxia Zhao\",\"doi\":\"10.1111/pbi.70274\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Ethylene plays a pivotal role during the fruit ripening process in tomato (<jats:styled-content style=\\\"fixed-case\\\"><jats:italic>Solanum lycopersicum</jats:italic></jats:styled-content>). Previously, we have reported that the tomato EIN2‐like protein YELLOW‐FRUITED TOMATO1 (YFT1), a core component in the ethylene signal transduction pathway, exerts a critical regulatory function in the tomato fruit ripening process. However, the molecular mechanism of YFT1‐mediated ethylene signalling during tomato ripening still remains largely unknown. In this study, we performed yeast two‐hybrid screens of a cDNA expression library from tomato fruits and obtained a new YFT1‐interacting protein, SlTFT1 (TOMATO FOURTEEN‐THREE PROTEIN 1), and confirmed their interaction in vitro and in vivo. Further bimolecular fluorescence complementation (BiFC) assays indicated that SlTFT1 can specifically bind the canonical and non‐canonical on the carboxyl terminus of YFT1 (YFT1‐C). Triple response assays and ethylene responsive gene expression analysis demonstrated that SlTFT1 positively regulates the ethylene signalling pathway in a YFT1‐dependent manner. Phenotypical and biochemical analysis of SlTFT1 overexpression lines and loss‐of‐function mutants showed that <jats:italic>SlTFT1</jats:italic> significantly accelerated ethylene emission, chromoplast development, lycopene accumulation and fruit ripening rates; however, the deletion of <jats:italic>YFT1</jats:italic> and/or <jats:italic>SlTFT1</jats:italic> lesion generated the opposite results, suggesting that <jats:italic>SlTFT1</jats:italic> positively regulates tomato fruit ripening also in a YFT1‐dependent manner. Co‐expression and western blotting assays showed that SlTFT1 can efficiently prevent YFT1 from protein degradation mediated by an F‐box protein SlETP2‐like3, suggesting that SlTFT1 can stabilise YFT1 through their physical interaction. Collectively, our results reveal a genetic and molecular framework of the SlTFT1‐YFT1 complex, which modulates tomato fruit ripening by regulating ethylene signalling.\",\"PeriodicalId\":221,\"journal\":{\"name\":\"Plant Biotechnology Journal\",\"volume\":\"91 1\",\"pages\":\"\"},\"PeriodicalIF\":10.5000,\"publicationDate\":\"2025-07-22\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Plant Biotechnology Journal\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1111/pbi.70274\",\"RegionNum\":1,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Plant Biotechnology Journal","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1111/pbi.70274","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
乙烯在番茄果实成熟过程中起着关键作用。在此之前,我们已经报道了番茄EIN2样蛋白YFT1 (YELLOW‐FRUITED TOMATO1)是乙烯信号转导通路的核心成分,在番茄果实成熟过程中发挥着关键的调控作用。然而,在番茄成熟过程中,YFT1介导的乙烯信号传导的分子机制仍然是未知的。在这项研究中,我们对来自番茄果实的cDNA表达文库进行了酵母双杂交筛选,获得了一个新的YFT1相互作用蛋白SlTFT1 (tomato FOURTEEN - THREE protein 1),并在体外和体内证实了它们的相互作用。进一步的双分子荧光互补(BiFC)实验表明,SlTFT1可以特异性结合YFT1羧基末端的典型和非典型蛋白(YFT1‐C)。三重应答实验和乙烯应答基因表达分析表明,SlTFT1以YFT1依赖性的方式正向调节乙烯信号通路。对SlTFT1过表达系和功能缺失突变体的表型和生化分析表明,SlTFT1显著加快了乙烯释放、叶绿体发育、番茄红素积累和果实成熟速度;然而,YFT1缺失和/或SlTFT1损伤产生相反的结果,这表明SlTFT1也以YFT1依赖的方式积极调节番茄果实成熟。共表达和western blotting实验表明,SlTFT1可以有效地阻止F - box蛋白SlETP2 - like3介导的YFT1蛋白降解,这表明SlTFT1可以通过它们的物理相互作用来稳定YFT1。总之,我们的研究结果揭示了SlTFT1 - YFT1复合物的遗传和分子框架,该复合物通过调节乙烯信号调节番茄果实成熟。
The 14‐3‐3 Protein SlTFT1 Accelerates Tomato Fruit Ripening by Binding and Stabilising YFT1 in the Ethylene Signalling Pathway
Ethylene plays a pivotal role during the fruit ripening process in tomato (Solanum lycopersicum). Previously, we have reported that the tomato EIN2‐like protein YELLOW‐FRUITED TOMATO1 (YFT1), a core component in the ethylene signal transduction pathway, exerts a critical regulatory function in the tomato fruit ripening process. However, the molecular mechanism of YFT1‐mediated ethylene signalling during tomato ripening still remains largely unknown. In this study, we performed yeast two‐hybrid screens of a cDNA expression library from tomato fruits and obtained a new YFT1‐interacting protein, SlTFT1 (TOMATO FOURTEEN‐THREE PROTEIN 1), and confirmed their interaction in vitro and in vivo. Further bimolecular fluorescence complementation (BiFC) assays indicated that SlTFT1 can specifically bind the canonical and non‐canonical on the carboxyl terminus of YFT1 (YFT1‐C). Triple response assays and ethylene responsive gene expression analysis demonstrated that SlTFT1 positively regulates the ethylene signalling pathway in a YFT1‐dependent manner. Phenotypical and biochemical analysis of SlTFT1 overexpression lines and loss‐of‐function mutants showed that SlTFT1 significantly accelerated ethylene emission, chromoplast development, lycopene accumulation and fruit ripening rates; however, the deletion of YFT1 and/or SlTFT1 lesion generated the opposite results, suggesting that SlTFT1 positively regulates tomato fruit ripening also in a YFT1‐dependent manner. Co‐expression and western blotting assays showed that SlTFT1 can efficiently prevent YFT1 from protein degradation mediated by an F‐box protein SlETP2‐like3, suggesting that SlTFT1 can stabilise YFT1 through their physical interaction. Collectively, our results reveal a genetic and molecular framework of the SlTFT1‐YFT1 complex, which modulates tomato fruit ripening by regulating ethylene signalling.
期刊介绍:
Plant Biotechnology Journal aspires to publish original research and insightful reviews of high impact, authored by prominent researchers in applied plant science. The journal places a special emphasis on molecular plant sciences and their practical applications through plant biotechnology. Our goal is to establish a platform for showcasing significant advances in the field, encompassing curiosity-driven studies with potential applications, strategic research in plant biotechnology, scientific analysis of crucial issues for the beneficial utilization of plant sciences, and assessments of the performance of plant biotechnology products in practical applications.