摘要
【目的】对近缘毛壳Chaetomium subaffine菌株LB-1产生抑菌物质的部位及提取方法进行研究,为开发生防菌株LB-1防治植物病害奠定基础。【方法】以番茄灰霉病菌Botrytis cinerea和玉米大斑病菌Exserohilum turcicum为指示菌,采用平板密封培养法检测菌株LB-1是否产生挥发性抑菌物质;采用菌体超声破碎和液态培养的方式检测菌株LB-1产生的非挥发性抑菌物质存在的部位;采用含毒培养基法和滤纸片法检测菌株LB-1培养液的硫酸铵沉淀、盐酸沉淀和有机溶剂萃取物的抑菌效果,以确定菌株LB-1培养液中抑菌物质的提取方法。【结果】菌株LB-1与2种供试病原菌密封共培养时,对病原菌的生长没有明显影响,表明菌株LB-1不能产生挥发性抑菌物质。菌株LB-1菌体胞内提取物的抑菌活性与对照无差异,但其培养液对B. cinerea和E. turcicum的生长有较强的抑制作用,表明菌株LB-1产生的抑菌物质存在于菌体细胞外。菌株LB-1培养液的硫酸铵沉淀和盐酸沉淀对2种供试植物病原真菌的生长均无抑制效果,但有机溶剂萃取可获得菌株LB-1培养液中的抑菌物质,其中正丁醇萃取物抑菌效果最好,当其质量浓度为0.1 mg/mL时,对B. cinerea和E. turcicum的生长抑制率分别高达59.80%和58.37%。【结论】菌株LB-1通过产生胞外非挥发性物质抑制植物病原真菌的生长,培养液中的抑菌物质可通过正丁醇萃取获得。
【Objective】The existing position and extraction method of antifungal substance produced by Chaetomium subaffine strain LB-1 were studied in order to lay a foundation for the development of the strain LB-1 to control plant diseases.【Method】With Botrytis cinerea and Exserohilum turcicum as test plant pathogens, the sealed plate assay was used to detect whether the strain LB-1 could produce volatile antifungal substance. The hyphae ultrasonic breaking and liquid culture methods were used to detect the existing position of nonvolatile antifungal substance produced by the strain LB-1. The extraction method of antifungal substance was determined by detecting the inhibitory effects of ammonium sulfate precipitation, hydrochloric acid precipitation and organic solvents extracts of strain LB-1 culture broth via poison plate assay and filter paper disc assay.【Result】The strain LB-1 did not have obvious inhibitory effect on the growth of the two test plant pathogens when being co-cultured in a sealed plate with each plant pathogen, indicating that strain LB-1 could not produce volatile antifungal substance. The antifungal activity of the intracellular extract of the strain LB-1 was not different from that of the control, but its culture broth had a strong inhibitory effect on B. cinerea and E.turcicum, indicating that the antifungal substances produced by the strain LB-1 existed outside the mycelium.Neither ammonium sulfate precipitate nor hydrochloric acid precipitate of strain LB-1 culture broth showed inhibitory effect on B. cinerea and E. turcicum, but the organic solvent extract of strain LB-1 culture broth showed antifungal effect, and the inhibition rate of n-butanol extract was the highest. When the concentration was 0.1 mg/mL, the inhibition rates against B. cinerea and E. turcicum growth were 59.80% and 58.37%respectively.【Conclusion】The strain LB-1 inhibited the growth of plant pathogenic fungi by producing extracellular nonvolatile antifungal substances, and the antifungal substance in the culture broth can be extracted by n-butanol.
作者
刘彩云
高秀清
赵静
LIU Caiyun;GAO Xiuqing;ZHAO Jing(College of Seed and Facility Agricultural Engineering,Weifang Universtiy,Weifang 261061,China)
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2023年第2期280-286,共7页
Journal of South China Agricultural University
基金
山东省自然科学基金(ZR2021MC144)。
关键词
近缘毛壳
抑菌物质
培养液
正丁醇粗提物
Chaetomium subaffine
Antifungal substance
Culture broth
Crude extract of n-butanol