Clathrin-mediated Endocytosis Facilitates Internalization of Magnaporthe oryzae Effectors into Rice Cells
Clathrin-mediated Endocytosis Facilitates Internalization of Magnaporthe oryzae Effectors into Rice Cells
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DOI：
10.1101/2021.12.28.474284
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发表时间：
2021-12
期刊：
影响因子：
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通讯作者：
Ely Oliveira-Garcia;Tej Man Tamang;Jungeun Park;Melinda Dalby;Magdalena Martin-Urdiroz;Clara Rodriguez Herrero;Annie Vu;Sunghun Park;N. Talbot;B. Valent
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文献类型：
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作者：
Ely Oliveira-Garcia;Tej Man Tamang;Jungeun Park;Melinda Dalby;Magdalena Martin-Urdiroz;Clara Rodriguez Herrero;Annie Vu;Sunghun Park;N. Talbot;B. Valent
真菌和卵菌将效应子传递到活的植物细胞中，以抑制防御并控制感染所需的植物过程。关于这些病原体将效应蛋白跨质膜转运到植物细胞质中的机制知之甚少。稻瘟病菌在易位之前将细胞质效应物分泌到专门的生物营养界面复合物（BIC）中。在这里，我们表明，BIC内的细胞质效应器被包装成囊泡，偶尔在宿主细胞质中观察到。荧光标记水稻的活细胞成像显示，效应囊泡与植物质膜和网格蛋白轻链-1（网格蛋白介导的内吞作用（CME）的标志物）共定位。使用病毒诱导的基因沉默（VIGS）和化学处理抑制CME导致缺乏囊泡的肿胀BIC中的细胞质效应物。与此相反，荧光标记物共定位，VIGS和化学抑制剂的研究未能支持网格蛋白独立的内吞作用在效应囊泡形成的主要作用。两个新的效应器，和已知的效应器CME抑制后的定位研究表明，细胞质效应器易位发生在附着胞下面入侵菌丝生长之前。综上所述，本研究提供的证据表明，细胞质效应易位是由网格蛋白介导的BIC内吞作用介导的，并表明M。植物内吞作用中的细胞因子。
Fungi and oomycetes deliver effectors into living plant cells to suppress defenses and control plant processes needed for infection. Little is known about the mechanism by which these pathogens translocate effector proteins across the plasma membrane into the plant cytoplasm. The blast fungus Magnaporthe oryzae secretes cytoplasmic effectors into a specialized biotrophic interfacial complex (BIC) before translocation. Here we show that cytoplasmic effectors within BICs are packaged into vesicles that are occasionally observed in the host cytoplasm. Live cell imaging with fluorescently-labeled rice showed that effector vesicles colocalize with plant plasma membrane and with clathrin light chain-1, a marker for clathrin-mediated endocytosis (CME). Inhibition of CME using Virus-Induced Gene Silencing (VIGS) and chemical treatments results in cytoplasmic effectors in swollen BICs lacking vesicles. In contrast, fluorescent marker co-localization, VIGS and chemical inhibitor studies failed to support a major role for clathrin-independent endocytosis in effector vesicle formation. Localization studies of two novel effectors, and of known effectors after CME inhibition, indicate that cytoplasmic effector translocation occurs underneath appressoria before invasive hyphal growth. Taken together, this study provides evidence that cytoplasmic effector translocation is mediated by clathrin-mediated endocytosis in BICs and suggests a role for M. oryzae effectors in co-opting plant endocytosis.