Abstract

BackgroundThe oomycete Phytophthora infestans causes late blight on potato and tomato. Despite extensive research, the P. infestans-host interaction is still poorly understood. To find new ways to further unravel this interaction we established a new infection system using MsK8 tomato cells. These cells grow in suspension and can be maintained as a stable cell line that is representative for tomato.ResultsMsK8 cells can host several Phytophthora species pathogenic on tomato. Species not pathogenic on tomato could not infect. Microscopy revealed that 16 h after inoculation up to 36% of the cells were infected. The majority were penetrated by a germ tube emerging from a cyst (i.e. primary infection) while other cells were already showing secondary infections including haustoria. In incompatible interactions, MsK8 cells showed defense responses, namely reactive oxygen species production and cell death leading to a halt in pathogen spread at the single cell level. In compatible interactions, several P. infestans genes, including RXLR effector genes, were expressed and in both, compatible and incompatible interactions tomato genes involved in defense were differentially expressed.ConclusionsOur results show that P. infestans can prosper as a pathogen in MsK8 cells; it not only infects, but also makes haustoria and sporulates, and it receives signals that activate gene expression. Moreover, MsK8 cells have the ability to support pathogen growth but also to defend themselves against infection in a similar way as whole plants. An advantage of MsK8 cells compared to leaves is the more synchronized infection, as all cells have an equal chance of being infected. Moreover, analyses and sampling of infected tissue can be performed in a non-destructive manner from early time points of infection onwards and as such the MsK8 infection system offers a potential platform for large-scale omics studies and activity screenings of inhibitory compounds.

Highlights

  • The oomycete Phytophthora infestans causes late blight on potato and tomato

  • Tomato cells growing in suspension can host Phytophthora infestans In order to investigate if MsK8 cells are susceptible to P. infestans, a fluorescent strain was used for inoculation and the infection process was monitored by microscopy (Additional file 1: Table S1)

  • Zoospores from P. infestans 14-3-GFP were mixed with MsK8 cells in a 1:1 ratio

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Summary

Introduction

The oomycete Phytophthora infestans causes late blight on potato and tomato. Despite extensive research, the P. infestans-host interaction is still poorly understood. To find new ways to further unravel this interaction we established a new infection system using MsK8 tomato cells. These cells grow in suspension and can be maintained as a stable cell line that is representative for tomato. In order to fend off pathogens and circumvent infection, plants have developed several defense mechanisms like cell wall thickening, reactive oxygen species (ROS) production and programmed cell death. These plant immune responses are often initiated by plasma membrane-spanning surface receptors as well as cytoplasmic receptors recognizing pathogen-derived molecules. Well-known examples of MAMPs are the bacterial flagellin, fungal chitin and oomycete elicitins such as Schoina et al Plant Methods (2017) 13:88

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