Mott G Adam, Thakur Shalabh, Smakowska Elwira, Wang Pauline W, Belkhadir Youssef, Desveaux Darrell, Guttman David S
Department of Cell & Systems Biology, University of Toronto, 25 Willcocks St., Toronto, Ontario, Canada.
Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna Biocenter (VBC), Dr Bohr Gasse 3, Vienna, 1030, Austria.
Genome Biol. 2016 May 9;17:98. doi: 10.1186/s13059-016-0955-7.
The recognition of microbe-associated molecular patterns during infection is central to the mounting of an effective immune response. In spite of their importance, it remains difficult to identify these molecules and the host receptors required for their perception, ultimately limiting our understanding of the role of these molecules in the evolution of host-pathogen relationships.
We employ a comparative genomics screen to identify six new immune eliciting peptides from the phytopathogenic bacterium Pseudomonas syringae. We then perform a reverse genetic screen to identify Arabidopsis thaliana leucine-rich repeat receptor-like kinases required for the recognition of these elicitors. We test the six elicitors on 187 receptor-like kinase knock-down insertion lines using a high-throughput peroxidase-based immune assay and identify multiple lines that show decreased immune responses to specific peptides. From this primary screen data, we focused on the interaction between the xup25 peptide from a bacterial xanthine/uracil permease and the Arabidopsis receptor-like kinase xanthine/uracil permease sensing 1; a family XII protein closely related to two well-characterized receptor-like kinases. We show that xup25 treatment increases pathogenesis-related gene induction, callose deposition, seedling growth inhibition, and resistance to virulent bacteria, all in a xanthine/uracil permease sensing 1-dependent manner. Finally, we show that this kinase-like receptor can bind the xup25 peptide directly. These results identify xup25 as a P. syringae microbe-associated molecular pattern and xanthine/uracil permease sensing 1 as a receptor-like kinase that detects the xup25 epitope to activate immune responses.
The present study demonstrates an efficient method to identify immune elicitors and the plant receptors responsible for their perception. Further exploration of these molecules will increase our understanding of plant-pathogen interactions and the basis for host specificity.
在感染过程中识别微生物相关分子模式对于产生有效的免疫反应至关重要。尽管它们很重要,但仍然难以鉴定这些分子以及识别它们所需的宿主受体,最终限制了我们对这些分子在宿主 - 病原体关系演变中作用的理解。
我们采用比较基因组学筛选方法,从植物致病细菌丁香假单胞菌中鉴定出六种新的免疫激发肽。然后,我们进行反向遗传筛选,以鉴定识别这些激发子所需的拟南芥富含亮氨酸重复序列的类受体激酶。我们使用基于高通量过氧化物酶的免疫测定法,在187个类受体激酶敲除插入系上测试这六种激发子,并鉴定出多个对特定肽免疫反应降低的品系。从这个初步筛选数据中,我们聚焦于来自细菌黄嘌呤/尿嘧啶通透酶的xup25肽与拟南芥类受体激酶黄嘌呤/尿嘧啶通透酶感应蛋白1之间的相互作用;该蛋白是与两个已充分表征的类受体激酶密切相关的XII家族蛋白。我们表明,xup25处理以黄嘌呤/尿嘧啶通透酶感应蛋白1依赖的方式增加病程相关基因的诱导、胼胝质沉积、幼苗生长抑制以及对致病细菌的抗性。最后,我们表明这种类激酶受体可以直接结合xup25肽。这些结果确定xup25为丁香假单胞菌的微生物相关分子模式,黄嘌呤/尿嘧啶通透酶感应蛋白1为检测xup25表位以激活免疫反应的类受体激酶。
本研究展示了一种鉴定免疫激发子及其负责识别的植物受体的有效方法。对这些分子的进一步探索将增进我们对植物 - 病原体相互作用以及宿主特异性基础的理解。