β-1, 3-glucan synthase (GLS) catalyzes the synthesis of cell wall core component β-1,3-glucan which is essential for fungal survival and virulence, and is the important target for antifungal agent development.To explore its gene function in the survival and infection process of Botrytis cinerea, this study primarily cloned the full-length sequence of β-1, 3-glucan synthase coding gene (BcGLS).Its total length was 7 137 bp by encoding 1 932 amino acids.Bioinformatic analysis indicated that BcGLS protein (BcGLSp) had a theoretical relative molecular weight of 220.05 kDa and a theoretical isoelectric point of 7.97.Subcellular prediction of BcGLSp was located in the cell membrane with 17 transmembrane regions, it had no signal peptide and was a non-secreted protein.BcGLSp was a hydrophilic protein with 176 phosphorylation sites.The results of conservative domain analysis revealed that BcGLSp contains one β-1,3-glucan synthase component and one β-1,3-glucan synthase FKS1 domain, belonging to glucan-synthase superfamily.Phylogenetic analysis displayed that BcGLS had close genetic relationship with those from plant pathogens such as Ciborinia camelliae and Erysiphe neolycopersici.Different abiotic stresses (NaCl、SDS、H2O2) significantly reduced Botrytis mycelial growth rate, and real-time PCR further showed that the expression of BcGLS was up-regulated under these stresses, indicating that the transcriptional expression of BcGLS was closely associated to environmental stresses.This study provides theoretical materials and practical basis for further research on the function of BcGLS gene in the survival and infection process of B.cinerea and the novel control strategies of B.cinerea.
[1] GARFINKEL A R.The history of Botrytis taxonomy, the rise of phylogenetics, and implications for species recognition[J].Phytopathology, 2021, 111(3):437-454.
[2] WILLIAMSON B, TUDZYNSKI B, TUDZYNSKI P, et al.Botrytis cinerea:The cause of gray mould disease[J].Molecular Plant Pathology, 2007, 8(5):561-580.
[3] POVEDA J, BARQUERO M, GONZÁLEZ-ANDRÉS F.Insight into the microbiological control strategies against Botrytis cinerea using systemic plant resistance activation[J].Agronomy, 2020, 10(11):1822.
[4] SHAO W Y, ZHAO Y F, MA Z H.Advances in understanding fungicide resistance in Botrytis cinerea in China[J].Phytopathology, 2021, 111(3):455-463.
[5] ADNAN M, HAMADA M S, HAHN M, et al.Fungicide resistance of Botrytis cinerea from strawberry to procymidone and zoxamide in Hubei, China[J].Phytopathology Research, 2019, 1:17.
[6] 刘西莉,苗建强,张灿.植物病原菌抗药性及其抗性治理策略[J].农药学学报, 2022, 24(5):921-936.
LIU X L, MIAO J Q, ZHANG C.Fungicide resistance and the management strategies[J].Chinese Journal of Pesticide Science, 2022, 24(5):921-936.
[7] CHEUNG N, TIAN L, LIU X R, et al.The destructive fungal pathogen Botrytis cinerea-Insights from genes studied with mutant analysis[J].Pathogens, 2020, 9(11):923.
[8] BI K, LIANG Y, MENGISTE T, et al.Killing softly:A roadmap of Botrytis cinerea pathogenicity[J].Trends in Plant Science, 2023, 28(2):211-222.
[9] GARCIA-RUBIO R, DE OLIVEIRA H C, RIVERA J, et al.The fungal cell wall:Candida, Cryptococcus, and Aspergillus species[J].Frontiers in Microbiology, 2020, 10:2993.
[10] LATGÉ J P.The cell wall:A carbohydrate armour for the fungal cell[J].Molecular Microbiology, 2007, 66(2):279-290.
[11] GEOGHEGAN I, STEINBERG G, GURR S.The role of the fungal cell wall in the infection of plants[J].Trends in Microbiology, 2017, 25 (12):957-967.
[12] GOW N A R, LATGE J P, MUNRO C A.The fungal cell wall:Structure, biosynthesis, and function[J].Microbiology Spectrum, 2017, 5(3):FUNK-0035-2016.
[13] ISHIHARA S, HIRATA A, NOGAMI S, et al.Homologous subunits of 1,3-beta-glucan synthase are important for spore wall assembly in Saccharomyces cerevisiae[J].Eukaryotic cell, 2007, 6(2):143-156.
[14] MARTÍN V, GARCÍA B, CARNERO E, et al.Bgs3p, a putative 1,3-beta-glucan synthase subunit, is required for cell wall assembly in Schizosaccharomyces pombe[J].Eukaryotic Cell, 2003, 2(1):159-169.
[15] CORTÉS J C G, CARNERO E, ISHIGURO J, et al.The novel fission yeast (1,3)-beta-D-glucan synthase catalytic subunit Bgs4p is essential during both cytokinesis and polarized growth[J].Journal of Cell Science, 2005, 118(Pt 1):157-174.
[16] CORTÉS J C G, KONOMI M, MARTINS I M, et al.The (1,3)-beta-D-glucan synthase subunit Bgs1p is responsible for the fission yeast primary septum formation[J].Molecular Microbiology, 2007, 65(1):201-217.
[17] MOUYNA I, HENRY C, DOERING T L, et al.Gene silencing with RNA interference in the human pathogenic fungus Aspergillus fumigatus[J].FEMS Microbiology Letters, 2004, 237(2):317-324.
[18] RUIZ-HERRERA J, ORTIZ-CASTELLANOS L.Cell wall glucans of fungi.A review[J].The Cell Surface, 2019, 5:100022.
[19] OLIVEIRA-GARCIA E, DEISING H B.Infection structure-specific expression of β-1,3-glucan synthase is essential for pathogenicity of Colletotrichum graminicola and evasion of β-glucan-triggered immunity in maize[J].The Plant Cell, 2013, 25(6):2356-2378.
[20] IBE C, MUNRO C A.Fungal cell wall:An underexploited target for antifungal therapies[J].PloS Pathogens, 2021, 17(4):e1009470.
[21] CORTÉS J C G, CURTO M Á, CARVALHO V S D, et al.The fungal cell wall as a target for the development of new antifungal therapies[J].Biotechnology Advances, 2019, 37(6):107352.
[22] THIYAGARAJAN K, LATINI A, CANTALE C, et al.Structural characterization of the DRF1 gene of Aegilops speltoides and comparison of its sequence with those of B and other Triticeae genomes[J].Euphytica, 2020, 216(10):152.
[23] CABIB E, ARROYO J.How carbohydrates sculpt cells:Chemical control of morphogenesis in the yeast cell wall[J].Nature Reviews Microbiology, 2013.11(9):648-655.
[24] CUI F J, WU X H, TAO T L, et al.Functions of a glucan synthase gene GFGLS in mycelium growth and polysaccharide production of Grifola frondose[J].Journal of Agricultural and Food Chemistry, 2019, 67(32):8875-8883.
[25] JIANG L H, LI X F, ZAN X Y, et al.The β-1,3-glucan synthase gene GFGLS2 plays major roles in mycelial growth and polysaccharide synthesis in Grifola frondosa[J].Applied Microbiology and Biotechnology, 2022, 106(2):563-578.
[26] FU X, ZAN X Y, SUN L, et al.Functional characterization and structural basis of the β-1,3-glucan synthase CMGLS from mushroom Cordyceps militaris[J].Journal of Agricultural and Food Chemistry, 2022, 70(28):8725-8737.
[27] HU X L, YANG P, CHAI C D, et al.Structural and mechanistic insights into fungal β-1,3-glucan synthase FKS1[J].Nature, 2023, 616(7955):190-198.
