Plants evoke innate immunity against microbial challenges upon recognition of pathogen-associated molecular patterns (PAMPs), such as fungal cell wall chitin. Nevertheless, pathogens may circumvent the host PAMP-triggered immunity. We previously reported that the ascomycete Magnaporthe oryzae, a famine-causing rice pathogen, masks cell wall surfaces with α-1,3-glucan during invasion. Here, we show that the surface α-1,3-glucan is indispensable for the successful infection of the fungus by interfering with the plant's defense mechanisms. The α-1,3-glucan synthase gene MgAGS1 was not essential for infectious structure development but was required for infection in M. oryzae. Lack or degradation of surface α-1,3-glucan increased fungal susceptibility towards chitinase, suggesting the protective role of α-1,3-glucan against plants' antifungal enzymes during infection. Furthermore, rice plants secreting bacterial α-1,3-glucanase (AGL-rice) showed strong resistance not only to M. oryzae but also to the phylogenetically distant ascomycete Cochlioborus miyabeanus and the polyphagous basidiomycete Rhizoctonia solani; the histocytochemical analysis of the latter two revealed that α-1,3-glucan also concealed cell wall chitin in an infection-specific manner. Treatment with α-1,3-glucanase in vitro caused fragmentation of infectious hyphae in R. solani but not in M. oryzae or C. miyabeanus, indicating that α-1,3-glucan is also involved in maintaining infectious structures in some fungi. Importantly, rapid defense responses were evoked (a few hours after inoculation) in the AGL-rice inoculated with M. oryzae, C. miyabeanus and R. solani as well as in non-transgenic rice inoculated with the ags1 mutant. Taken together, our results suggest that α-1,3-glucan protected the fungal cell wall from degradative enzymes secreted by plants even from the pre-penetration stage and interfered with the release of PAMPs to delay innate immune defense responses. Because α-1,3-glucan is nondegradable in plants, it is reasonable that many fungal plant pathogens utilize α-1,3-glucan in the innate immune evasion mechanism and some in maintaining the structures. Magnaporthe oryzae, Cochlioborus miyabeanus, and Rhizoctonia solani are the top three fungal pathogens that are responsible for devastating damage to the production of rice, a staple cereal for half of the world's population. These fungal pathogens infect host plants despite the plants' innate immunity, which is activated upon recognition of a conserved cell wall component in fungi, such as chitin. Fungal pathogens seem to have evading mechanism(s) against the host innate immunity; however, the mechanisms are still unclear. In this study, we discovered a novel mechanism that is commonly used by fungal pathogens to prevent host innate immunity. In this mechanism, fungal pathogens mask the cell wall surfaces with α-1,3-glucan, a polysaccharide that plants cannot degrade. In fact, a transgenic rice secreting a bacterial α-1,3-glucanase, which is able to remove α-1,3-glucan on the fungal surfaces, obtained strong resistance to all of those fungal pathogens. We also showed that plants rapidly activated defense responses against fungi (even before the fungal penetration) when α-1,3-glucan on the fungal surfaces were damaged or removed. Our study suggests that fungal surface α-1,3-glucan interferes with host immunity in many fungal pathogens and that α-1,3-glucan is a potential target for controlling various fungal diseases in plants.

Summary: Oligosaccharides derived from cell wall of fungal pathogens induce host primary immune responses. To understand fungal strategies circumventing the host plant immune responses, cell wall polysaccharide localization was investigated using fluorescent labels during infectious structure differentiation in the rice blast fungus Magnaporthe grisea. α‐1,3‐glucan was labelled only on appressoria developing on plastic surfaces, whereas it was detected on both germ tubes and appressoria on plant surfaces. Chitin, chitosan and β‐1,3‐glucan were detected on germ tubes and appressoria regardless of the substrate. Major polysaccharides labelled at accessible surface of infectious hyphae were α‐1,3‐glucan and chitosan, but after enzymatic digestion of α‐1,3‐glucan, β‐1,3‐glucan and chitin became detectable. Immunoelectron microscopic analysis showed α‐1,3‐glucan and β‐1,3‐glucan intermixed in the cell wall of infectious hyphae; however, α‐1,3‐glucan tended to be distributed farther from the fungal cell membrane. The fungal cell wall became more tolerant to chitinase digestion upon accumulation of α‐1,3‐glucan. Accumulation of α‐1,3‐glucan was dependent on the Mps1 MAP kinase pathway, which was activated by a plant wax derivative, 1,16‐hexadecanediol. Taken together, α‐1,3‐glucan spatially and functionally masks β‐1,3‐glucan and chitin in the cell wall of infectious hyphae. Thus, a dynamic change of composition of cell wall polysaccharides occurs during plant infection in M. grisea.

The APSES protein family includes important transcriptional regulators of morphological processes in ascomycetes. We identified a deletion mutant of the APSES protein Mstu1 in Magnaporthe grisea th...

α-1,3-Glucan, a component of the fungal cell wall, is a refractory polysaccharide for most plants. Previously, we showed that various fungal plant pathogens masked their cell wall surfaces with α-1,3-glucan to evade plant immunity. This surface accumulation of α-1,3-glucan was infection specific, suggesting that plant factors might induce its production in fungi. Through immunofluorescence observations of fungal cell walls, we found that carrot (Daucus carota) extract induced the accumulation of α-1,3-glucan on germlings in Colletotrichum fioriniae, a polyphagous fungal pathogen that causes anthracnose disease in various dicot plants. Bioassay-guided fractionation of carrot leaf extract successfully identified two active substances that caused α-1,3-glucan accumulation in this fungus: lutein, a carotenoid widely distributed in plants, and stigmasterol, a plant-specific membrane component. Lutein, which had a greater effect on C. fioriniae, also induced α-1,3-glucan accumulation in other Colletotrichum species and in the phylogenetically distant rice pathogen Cochliobolus miyabeanus, but not in the rice pathogen Magnaporthe oryzae belonging to the same phylogenetic subclass as Colletotrichum. Our results suggested that fungal plant pathogens reorganize their cell wall components in response to specific plant-derived compounds, which these pathogens may encounter during infection.