Promotion of Arabidopsis immune responses by a rhizosphere fungus via supply of pipecolic acid to plants and selective augment of phytoalexins

The ascomycete insect pathogenic fungi such as Metarhizium species have been demonstrated with the abilities to form the rhizosphere or endophytic relationships with different plants for nutrient exchanges.In this study,after the evident infeasibility of bacterial disease development in the boxed sterile soils,we established a hydroponic system for the gnotobiotic growth of Arabidopsis thaliana with the wild-type and transgenic strain of Metarhizium robertsii.The transgenic fungus could produce a high amount of pipecolic acid(PIP),a pivotal plant-immune-stimulating metabolite.Fungal inoculation experiments showed that M.robertsii could form a non-selective rhizosphere relationship with Arabidopsis.Similar to the PIP uptake by plants after exogenous application,PIP level increased in Col-0 and could be detected in the PIP-non-producing Arabidopsis mutant(ald1)after fungal inoculations,indicating that plants can absorb the PIP produced by fungi.The transgenic fungal strain had a better efficacy than the wild type to defend plants against the bacterial pathogen and aphid attacks.Contrary to ald1,fmo1 plants could not be boosted to resist bacterial infection after treatments.After fungal inoculations,the phytoalexins camalexin and aliphatic glucosinolate were selectively increased in Arabidopsis via both PIP-dependent and-independent ways.This study unveils the potential mechanism of the fungus-mediated beneficial promotion of plant immunity against biological stresses.The data also highlight the added values of M.robertsii to plants beyond the direct suppression of insect pest populations.

Identification of a key G-protein coupled receptor in mediating appressorium formation and fungal virulence against insects

Fungal G-protein coupled receptors(GPCRs)play essential roles in sensing environmental cues including host signals.The study of GPCR in mediating fungus-insect interactions is still limited.Here we report the evolution of GPCR genes encoded in the entomopathogenic Metarhizium species and found the expansion of Pth11-like GPCRs in the generalist species with a wide host range.By deletion of ten candidate genes MrGpr1–MrGpr10 selected from the six obtained subfamilies in the generalist M.robertsii,we found that each of them played a varied level of roles in mediating appressorium formation.In particular,deletion of MrGpr8 resulted in the failure of appressorium formation on different substrates and the loss of virulence during topical infection of insects but not during injection assays when compared with the wild-type(WT)strain.Further analysis revealed that disruption of MrGpr8 substantially impaired the nucleus translocation of the mitogen-activated protein kinase(MAPK)Mero-Fus3 but not the MAPK Mero-Slt2 during appressorium formation.We also found that the defect ofΔMrGpr8 could not be rescued with the addition of cyclic AMP for appressorium formation.Relative to the WT,differential expression of the selected genes have also been detected inΔMrGpr8.The results of this study may benefit the understanding of fungus-interactions mediated by GPCRs.