柑橘溃疡病菌效应蛋白PthA4诱导溃疡病症形成的分子机制研究

The pthA4 gene of Xanthomonas citri subsp. citri is one critical tal gene required for canker development in host plants. Most importantly, transient expression of pthA4 induces a canker-like symptom in citrus young leaves. According to the deciphered TALE-DNA molecular code, this project aims to identify the target genes in citrus plants via transcriptome and bioinformatics analysis. The specific binding between TALE and DNA is confirmed by Yeat one-hybridization, EMSA and promoter-GusA fusion experiments. The expressions of target genes are interfered in citrus plants to discover their biological roles in canker development. The symptoms on RNAi plants are fully characterized by inoculation with X. citri subsp. citri or transient expression of pthA4. The original purpose of this work is to explore citrus susceptible genes specifically recognized by PthA4, which will help to discover the right susceptible gene for further editing and finally to produce novel resistance citrus varieties in the future.

柑橘溃疡病菌的效应蛋白PthA4是引起病害发生的一个关键tale，并且单个PthA4即可诱导柑橘形成类似溃疡的症状。本项目通过转录组学和生物信息学分析，根据TALE-DNA结合的分子密码，筛选PthA4在柑橘植物中的靶基因；运用酵母单杂交、EMSA和启动子融合等实验，确认PthA4蛋白与靶基因启动子之间的特异性识别；利用反向遗传学方法，在柑橘中沉默靶基因的表达，接种溃疡病菌或瞬时表达pthA4基因，证明靶基因在溃疡症状形成中的生物学功能。本研究旨在发现柑橘植物中与pthA4对应识别的感病基因及其表达，为通过修饰感病基因获得抗病新品种提供准确的科学线索。

柑橘溃疡病菌的效应蛋白PthA4是引起病害发生的一个关键tal，并且单个PthA4即可诱导柑橘形成类似溃疡的症状。本项目通过转录组学和生物信息学分析，根据TAL蛋白-DNA结合的分子密码，筛选PthA4在柑橘植物中的靶基因；运用酵母单杂交、EMSA和启动子融合等实验，确认PthA4蛋白与靶基因启动子之间的特异性识别；利用反向遗传学方法，在柑橘中沉默靶基因的表达，接种溃疡病菌或瞬时表达pthA4基因，证明靶基因在溃疡症状形成中的生物学功能。本研究旨在发现柑橘植物中与pthA4对应识别的感病基因及其表达，为通过修饰感病基因获得抗病新品种提供准确的科学线索。