M88体育-明升M88体育讯(通讯员 王丽慧)近日,果蔬园艺作物种质创新与利用全国重点实验室、我校园艺林学学院别之龙教授团队和欧阳波教授团队合作研究成果以“Transcription factor SlWRKY50 enhances cold tolerance in tomato by activating the jasmonic acid signaling”为题在Plant Physiology在线发表。研究解析了转录因子SlWRKY50通过介导茉莉酸信号途径调控番茄低温响应的分子机制,为进一步研究WRKY转录因子的潜在机制和植物低温响应机制奠定了基础,同时也为未来耐低温番茄品种的选育提供了新的基因靶点。
番茄是一种对冷害极为敏感的喜温蔬菜,如何增强番茄植株耐冷性对于冬春季番茄设施栽培具有重要意义。课题组前期发现以野生型多毛番茄作为砧木嫁接栽培番茄能提高其耐冷性,茉莉酸在多毛番茄嫁接增强栽培番茄耐冷性中具有重要作用(研究结果已发表在Horticulture Research https://doi.org/10.1093/hr/uhac227),但其中具体调控茉莉酸生物合成的作用机制尚未明确。
图1 SlWRKY50通过介导茉莉酸信号途径调控番茄低温响应
研究发现,茉莉酸合成途径关键基因SlAOS是导致耐冷的多毛番茄和冷敏感的栽培番茄接穗中茉莉酸含量差异的关键因子。通过启动子元件预测,加权基因共表达网络分析及RT-qPCR定量实验,找到SlAOS的上游调控因子SlWRKY50。进一步通过酵母单杂交、LUC荧光信号检测、GUS活性分析及EMSA实验,明确了SlWRKY50能够直接结合到SlAOS的启动子上并正调控其转录。进一步通过病毒诱导的基因沉默和转基因技术进行验证,结果发现SlWRKY50正调控茉莉酸的生物合成,以此抵御低温胁迫。此外,通过药理学实验证明,对SlWRKY50超量植株施用JA抑制剂DIECA则负调控其耐冷性。通过以上证据说明SlWRKY50是通过介导茉莉酸信号增强番茄苗的耐冷性,此外,还发现茉莉酸信号的关键调控因子MYC2直接靶向SlWRKY50启动子正调控其转录,并通过JA缺失突变体和一系列生化实验验证了这一点。
综上所述,低温胁迫下SlWRKY50可转录激活JA合成酶SlAOS的表达,促进JA积累,进而通过MYC2-WRKY50通路诱导SlWRKY50的表达,建立自我放大反馈回路。积累的JA可以激活抗氧化酶活性,增强活性氧清除能力,最终提高番茄的耐冷性(图1)。
M88体育-明升M88体育园艺林学学院博士研究生王丽慧为论文第一作者,别之龙教授和欧阳波教授为论文共同通讯作者。中国科学院遗传与发育生物学研究所李传友研究员对此研究提供了突变体种质资源。本研究得到了国家重点研发计划、湖北省自然科学基金创新群体、湖北省重点研发计划、中央高校基本科研业务费专项基金等项目的资助。
审核人:别之龙
【英文摘要】
Tomato (Solanum lycopersicum) is among the most important vegetables across the world, but cold stress usually affects its yield and quality. The wild tomato species Solanum habrochaites is commonly utilized as rootstock for enhancing resistance against abiotic stresses in cultivated tomato, especially cold resistance. However, the underlying molecular mechanism remains unclear. In this research, we confirmed that S. habrochaites rootstock can improve the cold tolerance of cultivated tomato scions, as revealed by growth, physiological, and biochemical indicators. Furthermore, transcriptome profiling indicated significant differences in the scion of homo- and heterografted seedlings, including substantial changes in jasmonic acid (JA) biosynthesis and signaling, which were validated by RT–qPCR analysis. S. habrochaites plants had a high basal level of jasmonate, and cold stress caused a greater amount of active JA-isoleucine in S. habrochaites heterografts. Moreover, exogenous JA enhanced while JA inhibitor decreased the cold tolerance of tomato grafts. The JA biosynthesis-defective mutant spr8 also showed increased sensitivity to cold stress. All of these results demonstrated the significance of JA in the cold tolerance of grafted tomato seedlings with S. habrochaites rootstock, suggesting a future direction for the characterization of the natural variation involved in S. habrochaites rootstock-mediated cold tolerance.
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