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Involvement of Adapter Protein Complex 4 in Hypersensitive Cell Death Induced by Avirulent Bacteria

机译:适配器蛋白复合物4参与无毒细菌诱导的超敏细胞死亡。

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摘要

Plant immunity to avirulent bacterial pathogens is associated with subcellular membrane dynamics including fusion between the vacuolar and plasma membranes, resulting in hypersensitive cell death. Here, we report that ADAPTOR PROTEIN COMPLEX-4 (AP-4) subunits are involved in plant immunity associated with hypersensitive cell death. We isolated a mutant with a defect in resistance to an avirulent strain of Pseudomonas syringae pv. tomato (Pto) DC3000 avrRpm1 from a vacuolar protein sorting mutant library of Arabidopsis (Arabidopsis thaliana). The mutant was identical to gfs4-1, which has a mutation in the gene encoding the AP-4 subunit AP4B. Thus, we focused on AP4B and another subunit, AP4E. All of the mutants (ap4b-3, ap4b-4, ap4e-1, and ap4e-2) were defective in hypersensitive cell death and resistance to Pto DC3000 with the type III effector AvrRpm1 or AvrRpt2, both of which are recognized on the plasma membrane, while they showed slightly enhanced susceptibility to the type-III-secretion-deficient P. syringae strain hrcC. On the other hand, both ap4b-3 and ap4b-4 showed no defect in resistance to Pto DC3000 with the type III effector AvrRps4, which is recognized in the cytosol and does not induce hypersensitive cell death. Upon infection with Pto DC3000 avrRpt2, the ap4b-3 and ap4b-4 leaf cells did not show fusion between vacuolar and plasma membranes, whereas the wild-type leaf cells did. These results suggest that AP-4 contributes to cell death-associated immunity, possibly via membrane fusion, after type III effector-recognition on the plasma membrane.
机译:植物对无毒细菌病原体的免疫力与亚细胞膜动力学有关,包括液泡和质膜之间的融合,从而导致超敏细胞死亡。在这里,我们报告说,适配器蛋白复合物4(AP-4)亚基参与与超敏细胞死亡相关的植物免疫。我们分离出了一种突变体,该突变体对丁香假单胞菌无毒力菌株pv具有抗性。拟南芥(Arabidopsis thaliana)液泡蛋白分选突变体文库中的番茄(Pto)DC3000 avrRpm1。该突变体与gfs4-1相同,后者在编码AP-4亚基AP4B的基因中具有突变。因此,我们专注于AP4B和另一个子单元AP4E。所有突变体(ap4b-3,ap4b-4,ap4e-1和ap4e-2)在超敏性细胞死亡和对Pto DC3000的抗性方面均存在缺陷,并具有在血浆上均可识别的III型效应子AvrRpm1或AvrRpt2。膜,尽管它们显示出对III型分泌缺陷型丁香假单胞菌hrcC的敏感性略有增强。另一方面,ap4b-3和ap4b-4对III型效应子AvrRps4对Pto DC3000的抗药性均未显示缺陷,该缺陷在细胞质中被识别,并且不会引起超敏细胞死亡。用 Pto DC3000 avrRpt2 感染后, ap4b-3 ap4b-4 叶细胞之间没有融合液泡膜和质膜,而野生型叶细胞有。这些结果表明,AP-4在质膜上识别III型效应子后,可能通过膜融合促进了细胞死亡相关的免疫。

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