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针对水稻对稻瘟病菌抗性的靶向启动子编辑揭示了诱导SWEET14的转录激活样效应因子的不同活性。

Targeted promoter editing for rice resistance to Xanthomonas oryzae pv. oryzae reveals differential activities for SWEET14-inducing TAL effectors.

作者信息

Blanvillain-Baufumé Servane, Reschke Maik, Solé Montserrat, Auguy Florence, Doucoure Hinda, Szurek Boris, Meynard Donaldo, Portefaix Murielle, Cunnac Sébastien, Guiderdoni Emmanuel, Boch Jens, Koebnik Ralf

机构信息

UMR Interactions Plantes Microorganismes Environnement (IPME), IRD-CIRAD-Université, Montpellier, France.

Institut für Biologie, Institutsbereich Genetik, Martin-Luther-Universität Halle-Wittenberg, Halle (Saale), Germany.

出版信息

Plant Biotechnol J. 2017 Mar;15(3):306-317. doi: 10.1111/pbi.12613. Epub 2016 Dec 17.

DOI:10.1111/pbi.12613
PMID:27539813
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5316920/
Abstract

As a key virulence strategy to cause bacterial leaf blight, Xanthomonas oryzae pv. oryzae (Xoo) injects into the plant cell DNA-binding proteins called transcription activator-like effectors (TALEs) that bind to effector-binding elements (EBEs) in a sequence-specific manner, resulting in host gene induction. TALEs AvrXa7, PthXo3, TalC and Tal5, found in geographically distant Xoo strains, all target OsSWEET14, thus considered as a pivotal TALE target acting as major susceptibility factor during rice-Xoo interactions. Here, we report the generation of an allele library of the OsSWEET14 promoter through stable expression of TALE-nuclease (TALEN) constructs in rice. The susceptibility level of lines carrying mutations in AvrXa7, Tal5 or TalC EBEs was assessed. Plants edited in AvrXa7 or Tal5 EBEs were resistant to bacterial strains relying on the corresponding TALE. Surprisingly, although indels within TalC EBE prevented OsSWEET14 induction in response to BAI3 wild-type bacteria relying on TalC, loss of TalC responsiveness failed to confer resistance to this strain. The TalC EBE mutant line was, however, resistant to a strain expressing an artificial SWEET14-inducing TALE whose EBE was also edited in this line. This work offers the first set of alleles edited in TalC EBE and uncovers a distinct, broader range of activities for TalC compared to AvrXa7 or Tal5. We propose the existence of additional targets for TalC beyond SWEET14, suggesting that TALE-mediated plant susceptibility may result from induction of several, genetically redundant, host susceptibility genes by a single effector.

摘要

作为引起水稻白叶枯病的关键致病策略,水稻白叶枯病菌(Xanthomonas oryzae pv. oryzae,Xoo)向植物细胞中注射称为转录激活样效应因子(TALEs)的DNA结合蛋白,这些蛋白以序列特异性方式与效应因子结合元件(EBEs)结合,从而导致宿主基因的诱导。在地理上相距遥远的Xoo菌株中发现的TALEs AvrXa7、PthXo3、TalC和Tal5均靶向OsSWEET14,因此被认为是水稻与Xoo相互作用过程中作为主要感病因子的关键TALE靶点。在此,我们报告了通过在水稻中稳定表达TALE核酸酶(TALEN)构建体而产生的OsSWEET14启动子等位基因文库。评估了携带AvrXa7、Tal5或TalC EBE突变的株系的感病水平。在AvrXa7或Tal5 EBE中编辑的植物对依赖相应TALE的细菌菌株具有抗性。令人惊讶的是,尽管TalC EBE内的插入缺失阻止了OsSWEET14对依赖TalC的BAI3野生型细菌的诱导,但TalC反应性的丧失并未赋予对该菌株的抗性。然而,TalC EBE突变株系对表达人工SWEET14诱导TALE的菌株具有抗性,该菌株的EBE也在此株系中被编辑。这项工作提供了第一组在TalC EBE中编辑的等位基因,并揭示了与AvrXa7或Tal5相比,TalC具有独特的、更广泛的活性范围。我们提出TalC除了SWEET14之外还存在其他靶点,这表明TALE介导的植物感病性可能是由单个效应因子诱导多个遗传冗余的宿主感病基因所致。

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