三株无量山森林土壤芽孢杆菌鉴定及其生物活性挖掘  

Identification of Three Strains of Bacillus from the Forest Soil of Wuliang Mountain and Mining of Their Bioactivities

作　　者：王楠 廖永琴 施竹凤 申云鑫 杨童雨 冯路遥 矣小鹏 唐加菜 陈齐斌[1] 杨佩文[2] WANG Nan;LIAO Yong-qin;SHI Zhu-feng;SHEN Yun-xin;YANG Tong-yu;FENG Lu-yao;YI Xiao-peng;TANG Jia-cai;CHEN Qi-bin;YANG Pei-wen(College of Plant Protection,Yunnan Agricultural University,Kunming 650000;Agricultural Environment and Resources Institute,Yunnan Academy of Agricultural Sciences,Kunming 650205)

机构地区：[1]云南农业大学植物保护学院,昆明650000 [2]云南省农业科学院农业环境资源研究所,昆明650205

出　　处：《生物技术通报》2024年第2期277-288,共12页Biotechnology Bulletin

基　　金：云南省科技计划重大科技专项(202202AE090010,202202AE090015);国家自然科学基金项目(32060624);云南省专家基层科研工作站-杨佩文专家工作站(嵩明县,陆良县)(云人社通[2022]17号)。

摘　　要：【目的】挖掘无量山国家自然保护区潜在的高活性微生物菌株,旨在为农业绿色投入品研发提供高效的菌种资源。【方法】以番茄枯萎病菌(Fusarium oxysporum)为靶标筛选高拮抗活性菌株,观察对菌丝生长的影响,测定对孢子萌发的抑制作用;检测脂肽类化合物合成基因以及菌株体外产酶、解磷、解钾、固氮及产铁载体能力;结合形态学、生理生化特征和16S rRNA、gyrA、rpoB基因进行菌株鉴定,室内盆栽试验验证防病和促生效果。【结果】分离筛选获得153株可培养细菌,其中,菌株SH-53、N4471和N9456抑菌效果分别为92.35%、87.29%和88.47%,能抑制病原菌菌丝生长和分生孢子萌发,且对多种病原菌表现出良好拮抗活性。3株功能菌均具有溶锌、产淀粉酶和蛋白酶的能力,菌株SH-53和N4471还具有解磷、固氮、分泌纤维素酶以及产铁载体能力。3个功能菌基因组中均含有srfA、fenA、ituA、ituC、ituD、bymC等脂肽类化合物合成基因。经鉴定菌株SH-53为解淀粉芽孢杆菌(Bacillus amyloliquefaciens),N4471为卡式芽孢杆菌(B.cabrialesii),N9456为暹罗芽孢杆菌(B.siamensis)。盆栽试验结果表明,3株功能菌防效分别为84.66%、54.96%和59.74%,对番茄幼苗的株高、茎粗、根长等农艺性状均具有较好促进作用。【结论】3个菌株具有高效、广谱的抑菌活性,具有多样化的生物活性,作为高效的微生物资源具有广阔的应用前景。【Objective】In order to provide efficient microbial resource for the development of green agricultural inputs,we identified potential high-activity microbial strains from the National Wilderness Preservation System of Mount Wuliangshan.【Method】We used Fusarium oxysporum as a target to screen highly antagonistic strains,and observed the effect on hyphae growth and spore germination.We also tested the lipopeptide synthesis genes and the ability of the strain to produce enzymes,hydrolyze phosphorus,hydrolyze potassium,fix nitrogen and produce siderophore.In addition,the strains were identified according to their morphological,physiological and biochemical characteristics and 16S rRNA,gyrA and rpoB genes,and the effects of disease control and growth promotion were verified by pot experiment.【Result】A total of 153 strains of culturable bacteria were isolated and screened.Among them,the inhibition rates of SH-53,N4471 and N9456 were 92.35%,87.29%and 88.47%,respectively,it showed good antagonistic activity against many pathogens,and also inhibited the hypha growth and conidia germination of the pathogen.The three functional strains all had the abilities of dissolving zinc,producing amylase and protease,and the strains Sh-53 and N4471 also had the abilities of dissolving phosphorus,fixing nitrogen,secreting cellulase and producing siderophore.In addition,we found that srfA,fenA,ituA,ituC,ituD and bymC genes were found in the genomes of the three functional bacteria.Strain SH-53 was identified as Bacillus amyloliquefaciens,N4471 as B.cabrialesii,and N9456 as B.siamensis.The results of pot experiment showed that the control effects of the three functional bacteria were 84.66%,54.96%and 59.74%,respectively.They promoted the agronomic characters of tomato seedlings,such as plant height,stem diameter and root length.【Conclusion】As high-efficiency microbial resources,the three strains have broad-spectrum antibacterial activity,diverse biological activity,and have broad application prospects in the development of

关 键 词：解淀粉芽孢杆菌 卡式芽孢杆菌 暹罗芽孢杆菌 鉴定 生物活性 番茄枯萎病 促生 

分 类 号：S714.3[农业科学—林学]