TY - JOUR
T1 - Clathrin-mediated endocytosis facilitates the internalization of Magnaporthe oryzae effectors into rice cells
AU - Oliveira-Garcia, Ely
AU - Tamang, Tej Man
AU - Park, Jungeun
AU - Dalby, Melinda
AU - Martin-Urdiroz, Magdalena
AU - Herrero, Clara Rodriguez
AU - Vu, An Hong
AU - Park, Sunghun
AU - Talbot, Nicholas J.
AU - Valent, Barbara
N1 - Data availability statement: All study data are included in the article and/or supporting information. All strains and plasmids generated in this study are available from the authors upon request.
Funding Information: We acknowledge and thank Rick Nelson (Noble Foundation, Ardmore, OK) for providing the VIGS vectors and for hosting E.O.-G. in his laboratory to learn the VIGS assay. We thank the Talbot laboratory for assistance and hospitality during a working visit for E.O.-G at the University of Exeter. We thank Hiromasa Saitoh (Iwate Biotechnology Research Center, Kitakami, Iwate, Japan) and Ryohei Terauchi (Iwate Biotechnology Research Center, Kitakami, Iwate, and Kyoto University, Japan) for sharing transgenic rice expressing the plant plasma membrane marker Lti6B:GFP. We acknowledge Dr. David H. Burk and the Louisiana State University Shared Instrumentation Facility, for the help with confocal microscopy and imaging processing. This project was supported by Agriculture and Food Research Initiative Competitive Grant no. #2017-67013-26525 from the USDA National Institute of Food and Agriculture Plant Biotic Interactions program. Additional support came from Louisiana State University Agricultural Center Hatch project #LAB94477 and from Louisiana Board of Regents grant #LEQSF(2022-24)RD-A-01. This is Contribution no. 22-053-J from the Kansas Agricultural Experiment Station.
PY - 2023/7
Y1 - 2023/7
N2 - 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 punctate membranous effector compartments that are occasionally observed in the host cytoplasm. Live cell imaging with fluorescently labeled proteins in rice (Oryza sativa) showed that these effector puncta colocalize with the plant plasma membrane and with CLATHRIN LIGHT CHAIN 1, a component of clathrin-mediated endocytosis (CME). Inhibiting CME using virus-induced gene silencing and chemical treatments resulted in cytoplasmic effectors in swollen BICs lacking effector puncta. By contrast, fluorescent marker colocalization, gene silencing, and chemical inhibitor studies failed to support a major role for clathrin-independent endocytosis in effector translocation. Effector localization patterns indicated that cytoplasmic effector translocation occurs underneath appressoria before invasive hyphal growth. Taken together, this study provides evidence that cytoplasmic effector translocation is mediated by CME in BICs and suggests a role for M. oryzae effectors in coopting plant endocytosis.
AB - 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 punctate membranous effector compartments that are occasionally observed in the host cytoplasm. Live cell imaging with fluorescently labeled proteins in rice (Oryza sativa) showed that these effector puncta colocalize with the plant plasma membrane and with CLATHRIN LIGHT CHAIN 1, a component of clathrin-mediated endocytosis (CME). Inhibiting CME using virus-induced gene silencing and chemical treatments resulted in cytoplasmic effectors in swollen BICs lacking effector puncta. By contrast, fluorescent marker colocalization, gene silencing, and chemical inhibitor studies failed to support a major role for clathrin-independent endocytosis in effector translocation. Effector localization patterns indicated that cytoplasmic effector translocation occurs underneath appressoria before invasive hyphal growth. Taken together, this study provides evidence that cytoplasmic effector translocation is mediated by CME in BICs and suggests a role for M. oryzae effectors in coopting plant endocytosis.
UR - http://www.scopus.com/inward/record.url?scp=85161941130&partnerID=8YFLogxK
U2 - 10.1093/plcell/koad094
DO - 10.1093/plcell/koad094
M3 - Article
C2 - 36976907
AN - SCOPUS:85161941130
SN - 1040-4651
VL - 35
SP - 2527
EP - 2551
JO - Plant Cell
JF - Plant Cell
IS - 7
ER -