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Investigating the Role of Symbiosis-Regulated CAZymes in the Ectomycorrhizal Interaction between Laccaria bicolor and Populus

Investigating the Role of Symbiosis-Regulated CAZymes in the...

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多彩菌物美丽中国——中国菌物学会2019年学术年会

作者： ZHANG F LAbOUREL A CHAMPION C HAON M MACHADO E ANASONTZIS G KEMPPAINEN M PARDO A DAGUERRE Y DEVEAU A VENEAULT-FOURREY C KOHLER A ROSSO MN henrissat b bERRIN JG MARTIN F UMR 1136 INRA-Université de Lorraine ’Interactions Arbres/Microorganismes’ Laboratoire d’Excellence ARBRE Centre INRA-Lorraine State Key Laboratory of Grassland Agro-Ecosystems Institute of Innovation Ecology Lanzhou University UMR 1163 INRA-biodiversité et biotechnologie Fongiques Polytech Marseille Faculté des Sciences de Luminy Laboratorio de Micología Molecular Departamento de Ciencia y TecnologíaUniversidad Nacional de Quilmes CNRS UMR 7257 & Architecture et Fonction des Macromolécules Biologiques Aix-Marseille Université beijing Advanced Innovation Center for Tree breeding by Molecular Design Beijing Forestry University

In forest soils, ectomycorrhizal fungi establish a mutualistic symbiosis with tree roots. The mutualistic fungi trade host photoassimilates against soil nitrogen and phosphorus. Differentiation of symbiotic roots induces extensive cell wall architectural modifications in the host apoplastic space. The origin of enzymes involved in these cell wall modifications has been the subject of debate for several *** ectomycorrhizal basidiomycete Laccaria bicolor has a restricted set of carbohydrate-active enzymes(CAZymes) degrading plant cell wall polysaccharides. We speculate that several of the symbiosis-induced CAZymes are involved in the remodeling of the host apoplastic space. Here, we characterize the sole GH5 endoglucanase with a cellulose-binding motif(CbM1) domain(LbGH5) identified in the genome of L. *** showed that the LbGH5 gene is induced five-fold in ectomycorrhizal roots using qPCR and RNA-Seq;RNAi mutants with a decreased expression of LbGH5 have a lower ability to form ectomycorrhizal roots;the purified recombinant LbGH5 displayed enzymatic activity as endoglucanase;In situ localization of LbGH5 in ectomycorrhizal roots demonstrated that the enzyme accumulates in hyphal cell walls forming the mantle and Hartig net. In addition, the other highly induced polygalacturonase-LbGH28 was characterized. LbGH28 gene is upregulated five-fold in ectomycorrhizal roots by qPCR. The knock-down RNAi mutants are failed to form Hartig net in ectomycorrhizal roots. However, the overexpression of LbGH28 in *** is ability to induce more lateral roots and ectomycorrhiza comparing with wild type. We purified recombinant LbGH28 which showing polygalacturonase activity in vitro. LbGH28 is also secreted in cell wall of Hartig net and mantle by indirect immunofluorescence and immunogold *** together, these data suggest that ectomycorrhizal fungi secreted apoplastic effectors to remodeling plant cell wall for mycorrhizal formation.

关键词： Laccaria bicolor pectinase carbohydrate-active enzymes cell wall-modifying enzymes apoplastic effector endoglucanase polygalacturonase

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