特基拉芽孢杆菌WRN032对稻瘟病菌的生物防治潜力  

Biological Control Potential of Bacillus tequilensis WRN032 Against Magnaporthe oryzae

作　　者：蒋建兰[1,2] 陈玉 单潇潇 孙露 乔建军[1,2] 张桂山 Jiang Jianlan;Chen Yu;Shan Xiaoxiao;Sun Lu;Qiao Jianjun;Zhang Guishan(Key Laboratory of Systems Bioengineering(Ministry of Education),Tianjin University,Tianjin 300072,China;School of Chemical Engineering and Technology,Tianjin University,Tianjin 300072,China;Institute of Agricultural Resources and Regional Planning,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Key Laboratory of Microbial Resources Collection and Preservation,Ministry of Agriculture,Beijing 100081,China)

机构地区：[1]天津大学系统生物工程教育部重点实验室,天津300072 [2]天津大学化工学院,天津300072 [3]中国农业科学院农业资源与农业区划研究所,北京100081 [4]农业部农业微生物资源收集与保藏重点实验室,北京100081

出　　处：《天津大学学报（自然科学与工程技术版）》2023年第9期935-941,共7页Journal of Tianjin University：Science and Technology

基　　金：国家自然科学基金资助项目(31670113);国家重点研发计划资助项目(2017YFD0201401)。

摘　　要：植物病原真菌是农作物病害的主要致病菌,导致作物减产,严重阻碍农业发展.病原真菌种类繁多,作用机制复杂,防治难度大,传统化学防治手段无法满足当今发展需求,而生物防治是一种安全、绿色、高效的防控手段.以来源于土壤和植物根茎的多种细菌为研究对象,以菌种的抑菌活性为指标,采用牛津杯法,筛选出一株具有生物防治潜力的细菌WRN032,对黑腐皮壳菌(Rhizoctonia solani)、禾谷镰刀菌(Fusarium graminearum)和稻瘟病菌(Magnaporthe oryzae)的生长均具有不同程度的拮抗作用,其中对稻瘟病菌的抑制作用较强.使用石油醚、正丁醇和乙酸乙酯依次对菌株WRN032的发酵液进行萃取,发现其乙酸乙酯提取物对稻瘟病菌有强拮抗作用.结合生理生化鉴定及16S rDNA序列分析对其菌种进行鉴定,结果表明该细菌为特基拉芽孢杆菌(Bacillus tequilensis).对特基拉芽孢杆菌WRN032代谢产物中的活性物质进行稳定性试验,表明其在高温、强酸性、中性和弱碱性条件下均具有良好的稳定性,且对紫外光不敏感.采用柱层析法和半制备层析法对特基拉芽孢杆菌WRN032发酵液中的活性物质进行分离纯化,其纯化物对稻瘟病菌的相对抑制率与相同浓度的75%三环唑可湿性粉剂相近.当浓度为100.0μg/mL时,相对抑制率为51.2%,与75%三环唑可湿性粉剂(56.4%)基本相同.研究表明,特基拉芽孢杆菌WRN032具有作为农业生物防治剂的潜力.Plant pathogenic fungi are the main pathogens of crops that reduce crop yield and hinder agricultural development.Many kinds of pathogenic fungi have been identified with complex mechanisms of action leading to prevention and control difficulties.Traditional chemical control methods do not meet the needs of agriculture today,but biological control is a safe,green,and efficient means of prevention and control.We selected a variety of bacteria from soil and plant rhizomes as research objects and used the antibacterial activity of the bacteria as an index.The bacterial strain WRN032 with biological control potential was screened using the Oxford cup method.WRN032 has antagonistic effects on the growth of Rhizoctonia solani,Fusarium graminearum,and Magnaporthe oryzae,among which the inhibitory effect on M.oryzae was strong.The fermentation broth of strain WRN032 was extracted with petroleum ether,n-butanol,and ethyl acetate,in turn.The ethyl acetate extract of strain WRN032 had strong activ-ity against M.oryzae.The strain was identified using physiological and biochemical methods and 16S rDNA sequence analysis.The results showed that the bacterium was Bacillus tequilensis.A stability test of the active substance in the metabolites of B.tequilensis WRN032 revealed good stability under high temperature,strong acid,neutral,and weak alkali conditions and was not sensitive to ultraviolet light.The active substance in the B.tequilensis WRN032 fermentation broth was isolated and purified by column chromatography and semi-preparative chromatography.The relative inhibition rate of the purified products against rice blast was similar to that of 75%tricyclazole wettable pow-der at the same concentration.When the concentration was 100.0μg/mL,the relative inhibition rate was 51.2%,which was similar to the 75%for tricyclazole wettable powder(56.4%).This study shows that B.tequilensis WRN032 has potential as an agricultural biological control agent.

关 键 词：特基拉芽孢杆菌 WRN032 稻瘟病菌 分离鉴定 抑菌活性 生物防治 

分 类 号：Q939.9[生物学—微生物学]