Bacillus amyloliquefaciensBA-16-8所产fengycin对扩展青霉细胞的抑制机制研究
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摘要
目的:从细胞层面探究B.amyloliquefaciensBA-16-8所产fengycin对Penicillium expansum的抑制机制。方法:采用扫描电镜、透射电镜、核酸染色、光谱吸收和荧光显微观察分析fengycin对P.expansum细胞膜通透性及线粒体等细胞器的影响;采用凝胶阻滞实验探讨fengycin对菌丝体核酸的作用效果。结果:Fengycin处理P.expansum的菌丝体细胞,会导致细胞表面粗糙且细胞形态发生改变,细胞膜的完整性发生了改变,细胞结构遭到破坏。PI染色及光谱吸收实验结果显示,Fengycin可破坏细胞膜的完整性并增加P.expansum细胞膜的通透性。凝胶阻滞实验结果表明fengycin可与P.expansum的DNA于体外结合。结论:Fengycin可改变P.expansum的细胞膜通透性并作用于核酸,抑制基因的表达和细胞器的合成。
Objective: In order to explore theanti-fungal mechanism of fengycin produced by B. amyloliquefaciensBA-16-8 on P. expansum at the cell level. Methods: Scanning electron microscopy(SEM), transmission electron microscopy(TEM), PI nucleic acid fluorescent dye(PNCFD), fluorescence microscopy(FM) and spectral absorption was performed to investigate the effect of fengycin on the cellular morphological characteristics and structure of P. expansum mycelium. Electrophoretic mobility shift assays(EMSA) was performed toexamine the effect of fengycin on the nucleic acid of P. expansum. Results: SEM and TEM results revealed that the surface and intracellular protoplasm of P. expansum mycelium became roughand the morphological structure of the fungal cells were alteredafter with fengycin treatments, which meant fengycin could damage the cell membrane integrity and cell structure. The result of EMSA indicated that fengycin could combine with nonspecific P. expansum DNA in vitro. Conclusion: Fengycin could alter the cell membrane integrity and combined with nucleic acid such as DNA, which could inhibit gene expression and organelle synthesis.
Objective: In order to explore theanti-fungal mechanism of fengycin produced by B. amyloliquefaciensBA-16-8 on P. expansum at the cell level. Methods: Scanning electron microscopy(SEM), transmission electron microscopy(TEM), PI nucleic acid fluorescent dye(PNCFD), fluorescence microscopy(FM) and spectral absorption was performed to investigate the effect of fengycin on the cellular morphological characteristics and structure of P. expansum mycelium. Electrophoretic mobility shift assays(EMSA) was performed toexamine the effect of fengycin on the nucleic acid of P. expansum. Results: SEM and TEM results revealed that the surface and intracellular protoplasm of P. expansum mycelium became roughand the morphological structure of the fungal cells were alteredafter with fengycin treatments, which meant fengycin could damage the cell membrane integrity and cell structure. The result of EMSA indicated that fengycin could combine with nonspecific P. expansum DNA in vitro. Conclusion: Fengycin could alter the cell membrane integrity and combined with nucleic acid such as DNA, which could inhibit gene expression and organelle synthesis.
引文
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[3]Crossley A.A versatile multi-specimen holder for processing and critical point drying of materials for examination in the scanning electron microscope(SEM)[J].Journal of Microscopy,1976,108(3):349-352
[4]Mohammadi B,Salouti M.Extracellular bioynthesis of silver nanoparticles by Penicillium chrysogenum and Penicillium expansum[J].Synthesis and Reactivity in Inorganic,Metal-Organic,and Nano-Metal Chemistry,2015,45(6):844-847
[5]付瑞敏,邢文会,张红,等.扩展青霉拮抗菌的选育及抑菌机制初探[J].食品工业科技,2016,(5):154-158
[6]付瑞敏,于烽,常慧萍,等.一株扩展青霉拮抗菌的分离、鉴定及抑菌活性物质研究[J].微生物学通报,2016,43(8):1-10
[7]Zhao Y,Odhiambo B O,Qiu J,et al.Potential of natamycin in enhancing the antagonistic activity of Bamyloliquefaciens BGP20 against post-harvest bacterial soft rot of green pepper[J].Biocontrol Science and Technology,2016,26(3):402-413
[8]Fracchia L,Ceresa C,Franzetti A,et al.Indu-strial applications of biosurfactants[J].Biosurfactants:Production and Utilization-Processes,Technologies,and Economics,2014,159(1):245-245
[9]Wachenforff-Neumann U,Anadersch W,Stenzel K,et al.Composition comprising a biological control agent and a fungicide selected from inhibitors of the ergosterol biosynthesis[P].US Patent Application:14/404,023.2013-05-29
[10]Wachendorff-Neumann U,Andersch W,Springer B,et al.Composition comprising a biological control agent and a fungicide selected from inhibitors of amino acid or protein biosynthesis and inhibitors of cell wall synthesis[P].US Patent Application:14/318,130.2014-06-27
[11]Beld J,Sonnenschein E C,Vickery C R,et al.The phosphopantetheinyl transferases:catalysis of a posttranslational modification crucial for life[J].Natural product reports,2014,31(1):61-108
[12]Tanaka K,Ishihara A,Nakajima H.Isolation of anteisoC17,iso-C17,iso-C16,and iso-C15 Bacillomycin D from Bamyloliquefaciens SD-32 and their antifungal activities against plant pathogens[J].Journal of agricultural and food chemistry,2014,62(7):1469-1476
[13]姚磊,徐良雄,薛璟花,等.枝顶孢属真菌的抑菌活性及其代谢产物研究[J].热带亚热带植物学报,2012,20(1):192-196
[14]Mohammadi B,Salouti M.Extracellular bioynthesis of silver nanoparticles by Penicillium chrysogenum and Pexpansum[J].Synthesis and Reactivity in Inorganic,Metal-Organic,and Nano-Metal Chemistry,2015,45(6):844-847
[15]Parvu M,Parvu AE.Antifungal plant extracts science against microbial pathogens:comunicating current research and technological advances[J].Microbiology,2011,3(1):1055-1062
[16]Wachendorff-Neumann U,Andersch W,Springer B,et al.Composition comprising a biological control agent and a fungicide selected from inhibitors of amino acid or protein biosynthesis and inhibitors of cell wall synthesis[J].Patent Application,2014,318(1):130-131
[17]Fracchia L,Ceresa C,Franzetti A,et al.Production and utilization-processes,technologies,and economics[J].Industrial Applications of Biosurfactants,2014,159(1):245-246
[18]Alvarez F,Castro M,Príncipe A,et al.The planta ssociated Bacillusamy lolique faciens strains MEP218 and ARP23 capable of producing the cyclic lipopeptidesiturin or surfactin and fengycin are effective in biocontrol of sclerotinia stem rot disease[J].Journal of Applied Microbiology,2012,112(1):159-174
[19]Crossley A.A versatile multi-specimen holder for processing and critical point drying of materials for examination in the scanning electron microscope(SEM)[J].Journal of Microscopy,1976,108(3):349-352
[20]Mohammadi B.Salouti M.Extracellular bioynthesis of silver nanoparticles by Penicillium chrysogenum and Penicillium expansum[J].Synthesis and Reactivity in Inorganic,Metal-Organic,and Nano-Metal Chemistry,2015,45(6):844-847