Cunnac Sébastien, Wilson Ariane, Nuwer Jamie, Kirik Angela, Baranage Gayathri, Mudgett Mary Beth
Department of Biological Sciences, Stanford University, Stanford, California 94305, USA.
Plant Cell. 2007 Feb;19(2):688-705. doi: 10.1105/tpc.106.048710. Epub 2007 Feb 9.
AvrBsT is a type III effector from Xanthomonas campestris pv vesicatoria that is translocated into plant cells during infection. AvrBsT is predicted to encode a Cys protease that targets intracellular host proteins. To dissect AvrBsT function and recognition in Arabidopsis thaliana, 71 ecotypes were screened to identify lines that elicit an AvrBsT-dependent hypersensitive response (HR) after Xanthomonas campestris pv campestris (Xcc) infection. The HR was observed only in the Pi-0 ecotype infected with Xcc strain 8004 expressing AvrBsT. To create a robust pathosystem to study AvrBsT immunity in Arabidopsis, the foliar pathogen Pseudomonas syringae pv tomato (Pst) strain DC3000 was engineered to translocate AvrBsT into Arabidopsis by the Pseudomonas type III secretion (T3S) system. Pi-0 leaves infected with Pst DC3000 expressing a Pst T3S signal fused to AvrBsT-HA (AvrBsTHYB-HA) elicited HR and limited pathogen growth, confirming that the HR leads to defense. Resistance in Pi-0 is caused by a recessive mutation predicted to inactivate a carboxylesterase known to hydrolyze lysophospholipids and acylated proteins in eukaryotes. Transgenic Pi-0 plants expressing the wild-type Columbia allele are susceptible to Pst DC3000 AvrBsTHYB-HA infection. Furthermore, wild-type recombinant protein cleaves synthetic p-nitrophenyl ester substrates in vitro. These data indicate that the carboxylesterase inhibits AvrBsT-triggered phenotypes in Arabidopsis. Here, we present the cloning and characterization of the SUPPRESSOR OF AVRBST-ELICITED RESISTANCE1.
AvrBsT是野油菜黄单胞菌致病变种(Xanthomonas campestris pv vesicatoria)的一种III型效应蛋白,在感染过程中会被转运到植物细胞中。预测AvrBsT编码一种靶向细胞内宿主蛋白的半胱氨酸蛋白酶。为了剖析AvrBsT在拟南芥中的功能和识别机制,对71个生态型进行了筛选,以鉴定在野油菜黄单胞菌野油菜致病变种(Xanthomonas campestris pv campestris,Xcc)感染后引发AvrBsT依赖性过敏反应(HR)的株系。仅在感染了表达AvrBsT的Xcc菌株8004的Pi-0生态型中观察到了HR。为了创建一个强大的病理系统来研究拟南芥中AvrBsT介导的免疫,对叶部病原菌丁香假单胞菌番茄致病变种(Pseudomonas syringae pv tomato,Pst)菌株DC3000进行改造,使其通过假单胞菌III型分泌(T3S)系统将AvrBsT转运到拟南芥中。感染了表达与AvrBsT-HA融合的Pst T3S信号(AvrBsTHYB-HA) 的Pst DC3000的Pi-0叶片引发了HR并限制了病原菌的生长,证实HR导致了防御反应。Pi-0中的抗性是由一个隐性突变引起的,该突变预计会使一种已知在真核生物中水解溶血磷脂和酰化蛋白的羧酸酯酶失活。表达野生型哥伦比亚等位基因的转基因Pi-0植物对Pst DC3000 AvrBsTHYB-HA感染敏感。此外,野生型重组蛋白在体外可切割合成的对硝基苯酯底物。这些数据表明羧酸酯酶抑制了拟南芥中AvrBsT触发的表型。在此,我们展示了AvrBsT引发抗性的抑制因子1(SUPPRESSOR OF AVRBST-ELICITED RESISTANCE1)的克隆和特性分析。