MARVEL-ous extracellular vesicles carry RXLR effectors into host plant cells
Lead Research Organisation:
James Hutton Institute
Department Name: Cell & Molecular Sciences
Abstract
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Technical Summary
Oomycete plant pathogens cause severe damage to major food crops. The potato and tomato late blight pathogen Phytophthora infestans causes annual global losses estimated to exceed $10 bn. Late blight disease is driven by effector proteins, secreted by P. infestans, that act on (apoplastic effectors) or within (cytoplasmic effectors) host cells to facilitate infection. There is a considerable gap in our knowledge of how cytoplasmic RXLR effectors are secreted from P. infestans and delivered into plant cells. We have generated unpublished data showing that RXLR effectors associate with extracellular vesicles (EVs). The EV proteome revealed proteins containing MARVEL domains containing multiple transmembrane helices which co-associate with RXLR Pi04314 in EVs.
In our proposal we aim to confirm whether MARVEL proteins are markers of EVs associated with RXLRs and use cutting-edge super-resolution microscopy to provide a detailed and deep investigation of their secretion and delivery at the haustorial interface between P. infestans and its hosts. We will remove the MARVEL proteins by gene silencing and CRISPR/cas12 knockout to determine their importance to effector secretion and thus to infection. Finally, we will use the MARVEL proteins to immunopurify EVs following in vitro growth and during infection and study the EV proteome to identify factors involved in biogenesis, structure and function of EVs, and to determine the cargo of virulence proteins delivered to hosts during infection.
This work will provide a deep understanding of a key fundamental effector delivery mechanism likely to be shared across oomycete pathogens which can be targeted by agrochemical or biotechnological means to prevent crop disease.
In our proposal we aim to confirm whether MARVEL proteins are markers of EVs associated with RXLRs and use cutting-edge super-resolution microscopy to provide a detailed and deep investigation of their secretion and delivery at the haustorial interface between P. infestans and its hosts. We will remove the MARVEL proteins by gene silencing and CRISPR/cas12 knockout to determine their importance to effector secretion and thus to infection. Finally, we will use the MARVEL proteins to immunopurify EVs following in vitro growth and during infection and study the EV proteome to identify factors involved in biogenesis, structure and function of EVs, and to determine the cargo of virulence proteins delivered to hosts during infection.
This work will provide a deep understanding of a key fundamental effector delivery mechanism likely to be shared across oomycete pathogens which can be targeted by agrochemical or biotechnological means to prevent crop disease.
