Marchantia polymorpha model reveals conserved infection mechanisms in the vascular wilt fungal pathogen Fusarium oxysporum

Abstract

How co-evolution has shaped the interaction between plants andtheir associated microbes remains a central question in organis-mic interactions (Bonfante & Genre, 2010; Delaux & Schor-nack, 2021). Plants have evolved a sophisticated and multilayeredimmune system to ward off potential microbial invaders (Jones& Dangl, 2006; Boller & Felix, 2009). In addition, pathogenshave developed mechanisms allowing them to enter living plants,colonise their tissues and overcome their defence responses.Pathogenicity factors can be either broadly conserved or speciesspecific and include regulators of cell signalling, gene expressionor development, as well as secreted effector molecules that modu-late the host environment (Jongeet al., 2011; Turr aet al., 2014;Weiberget al., 2014; Prestiet al., 2015; Ryder & Talbot, 2015;van der Does & Rep, 2017).A particularly destructive group of plant pathogens are thosecausing vascular wilt diseases, which infect the roots and colonisethe highly protected and nutrient poor niche of the xylem(Yadeta & Thomma, 2013). The ascomycete fungusFusariumoxysporum(Fo) represents a species complex with worldwidedistribution that provokes devastating losses in more than 150different crops (Deanet al., 2012). Fo exhibits a hemibiotrophlifestyle with an initial biotrophic phase characterised by intercel-lular growth in the root cortex, followed by invasion of the vascu-lature and transition to the necrotrophic phase resulting inmaceration and death of the colonised host (Redkaret al., 2021).In the soil, Fo is able to locate roots by sensing secreted plant per-oxidases via its sex pheromone receptors and the cell wallintegrity mitogen activated protein kinase (MAPK) pathway(Turr aet al., 2015). Once inside the root, the fungus secretes asmall regulatory peptide that mimics plant Rapid ALkalinisationFactor (RALF) to induce host alkalisation, which in turn activatesa conserved MAPK cascade that promotes plant invasive growth(Masachiset al., 2016). Additional pathogenicity determinantsinclude transcriptional regulators, fungus/plant cell wall remod-elling components or secondary metabolites, among others(Michielse & Rep, 2009).Individual Fo isolates exhibit host-specific pathogenicity,which is determined by lineage-specific (LS) chromosomes thatencode distinct repertoires of effectors known as Secreted inXylem (Six) (Maet al., 2010; van Damet al., 2016). Some Six proteins appear to primarily target plant defence responses, butcan also be recognised as avirulence factors by specific host recep-tors (Houtermanet al., 2009; Tintoret al., 2020). In addition tothe pathogenic forms, the Fo species complex (FOSC) alsoincludes endophytic isolates such as Fo47, which was isolatedfrom a natural disease suppressive soil (Alabouvette, 1986; Wanget al., 2020). Fo47 colonises plant roots without causing wiltsymptoms and functions as a biological control agent againstpathogenic Fo strains. How vascular wilt fungi such as Fo gainedthe ability to associate with plant hosts and evolved endophyticand pathogenic lifestyles remains poorly understood.The bryophyteMarchantia polymorpha(Mp) belongs to theancient lineage of liverworts and has emerged as the primenonvascular plant model for studying the evolution of molecularplant–microbe interactions (Evo-MPMI), due to its low geneticredundancy, the simplicity of its gene families and an accessiblemolecular genetic toolbox (Ishizakiet al., 2008; Lockhart, 2015;Bowmanet al., 2017; Upsonet al., 2018; Gimenez-Ibanezet al.,2019). Importantly, Mp possesses receptor-like kinases (RLKs),nucleotide binding, leucine-rich repeat receptors (NLRs) and sal-icylic acid (SA) pathway genes similar to those mediatingimmune signalling in angiosperms (Xueet al., 2012; Bowmanet al., 2017), therefore allowing the study of plant–microbe inter-actions across evolutionarily distant land plant lineages such asliverworts and eudicots, which diverged>450 million years ago(Ma) (Carellaet al., 2018). A current shortcoming of Mp is thatonly few pathogen infection models have been developed forin vitropathogenicity assays. These include the fungiXylariacubensisandColletotrichum sp1, the oomycetePhytophthorapalmivoraand the Gram-negative bacteriumPseudomonassyringae(Nelsonet al., 2018; Carellaet al., 2019; Gimenez-Ibanezet al., 2019). A survey of the Mp microbiome identified anumber of fungal endophytes, some of which can also act aspathogens (Matsuiet al., 2019; Nelson & Shaw, 2019). Whetherroot-infecting vascular wilt fungi can colonise this land plant lin-eage, which is evolutionarily distant to eudicots and lacks bothtrue roots and xylem, is currently unknown.Here we established a new pathosystem between Fo and Mp.We find that Fo isolates that are either endophytic or pathogenicon different crops (tomato, banana, cotton) are all able tocolonise and macerate the thallus of this nonvascular plant. Infec-tion of Mp by Fo requires fungal core pathogenicity factors,whereas LS effectors are dispensable suggesting that this vascularwilt fungus employs conserved mechanisms during infection ofevolutionarily distant plant lineages. We further show that thefungal transition from biotrophic intercellular growth tonecrotrophic maceration and sporulation, which on angiospermsrelies on host-specific factors promoting xylem invasion, occursdirectly on the nonvascular plant Mp.