三株酚酸降解菌的筛选与鉴定及其生物活性  

Three phenolic acid-degrading bacterial strains:screening,identification,and biological activities

作　　者：廖永琴 王楠 申云鑫 唐加菜 施竹凤 矣小鹏 冯路遥 李铭刚[3] 何永宏[1] 杨佩文[2] LIAO Yongqin;WANG Nan;SHEN Yunxin;TANG Jiacai;SHI Zhufeng;YI Xiaopeng;FENG Luyao;LI Minggang;HE Yonghong;YANG Peiwen(College of Plant Protection,Yunnan Agricultural University,Kunming 650000,Yunnan,China;Agricultural Environment and Resources Institute,Yunnan Academy of Agricultural Sciences,Kunming 650205,Yunnan,China;Resource Plant Research Institute,Yunnan University,Kunming 650091,Yunnan,China)

机构地区：[1]云南农业大学植物保护学院,云南昆明650000 [2]云南省农业科学院农业环境资源研究所,云南昆明650205 [3]云南大学资源植物研究院,云南昆明650091

出　　处：《微生物学通报》2024年第6期2193-2214,共22页Microbiology China

基　　金：云南省科技计划重大科技专项(202202AE090010,202202AE090015);云南省专家基层科研工作站——杨佩文专家工作站(云人社通[2022]17号)。

摘　　要：【背景】常见作物连续种植后会导致根系分泌酚酸类自毒物质,该类物质是引起土传病害和连作障碍频发的主要因素。【目的】探讨利用有益微生物降解作物根系分泌物苯甲酸,缓解作物土传病害发生,并储备有益菌种资源。【方法】通过以苯甲酸为唯一碳源的无机盐培养基筛选无量山菌株资源,结合高效液相色谱仪测定酚酸降解菌株对苯甲酸的降解率;利用平板对峙试验,并基于缓解苯甲酸抑制番茄种子萌发试验及番茄幼苗促生长和对番茄枯萎病防效试验验证菌株的生物活性;结合形态学、生理生化和16S rRNA基因序列对菌株进行分类学鉴定。【结果】从无量山筛选得到4株放线菌和3株细菌具有酚酸降解能力,酚酸降解菌株在摇瓶培养72 h时对苯甲酸降解率分别为100%、99.83%、99.89%、99.87%、64.91%、56.92%和49.33%;其中2株放线菌YNK-FS0018、YNK-FS0019和一株细菌YNK-FB0022能较好地利用苯甲酸。番茄种子萌发试验结果表明菌株YNK-FS0018、YNK-FS0019和YNK-FB0022均可以缓解苯甲酸对种子萌发的抑制,促进种子萌发,萌发率分别为73%、73%和97%,并促进种子根生长,平均根长为13.31、13.04和14.56 cm;平板对峙试验结果表明菌株YNK-FS0018、YNK-FS0019和YNK-FB0022均对病原菌有广谱抗性;抗生素基因检测结果为菌株YNK-FS0018、YNK-FS0019具有PKSII、NRPS基因,菌株YNK-FB0022具有fenD、bioA、yndJ、ysnE、ituC、sboA和srfAB基因;盆栽试验结果显示菌株YNK-FS0018对番茄株高、地上鲜重、地上干重、根重、根长和茎粗有促生作用,各项分别增加53.40%、67.50%、57.50%、73.50%、77.28%和9.40%;菌株YNK-FS0019处理的株高、地上鲜重、地上干重、根重、根长和茎粗分别增加23%、40.90%、14.00%、57.00%、26.30%和16.80%;菌株YNK-FB0022处理的株高、地上鲜重、地上干重、根重、根长和茎粗分别增加24.50%、44.70%、26.40%、75.10%、98.00%和9.00%;3株菌对番茄枯[Background]Continuous planting of common crops can lead to the secretion of phenolic acid autotoxic substances in roots,which are the main factors causing soil-borne diseases and continuous cropping obstacles.[Objective]To mine the microorganisms capable of degrading benzoic acid secreted by crop roots,reduce the occurrence of soil-borne diseases,and reserve beneficial microbial resources.[Methods]We used benzoic acid as the sole carbon source in the inorganic salt medium to screen the strain resources of Wuliang Mountain and determined the degradation rate of benzoic acid by high-performance liquid chromatography.The plate confrontation test,tomato seed germination experiment,tomato seedling growth experiment,and pathogen(Fusarium wilt)inoculation experiment with tomato seedlings were carried out to examine the biological activities of the strains.The strains were identified based on morphological,physiological,and biochemical characteristics and 16S rRNA gene sequences.[Results]Four strains of Actinomycetes and three strains of bacteria with phenolic acid-degrading ability were screened from Wuliang Mountain.After culture in shake flasks for 72 h,the strains showed the benzoic acid-degrading rate of 100%,99.83%,99.89%,99.87%,64.91%,56.92%,and 49.33%,respectively.Two Actinomycetes strains(YNK-FS0018 and YNK-FS0019)and one bacterial strain(YNK-FB0022)could make good use of benzoic acid.The treatments with the three strains alleviated the inhibitory effect of benzoic acid on tomato seed germination,with the seed germination rates of 73%,73%,and 97%,the root length of 13.31,13.04,and 14.56 cm,respectively.Furthermore,the three strains had broad-spectrum resistance to pathogens.YNK-FS0018 and YNK-FS0019 carried PKSII and NRPS,and YNK-FB0022 carried fenD,bioA,yndJ,ysnE,ituC,sboA,and srfAB.The results of pot experiments showed that strain YNK-FS0018 increased the plant height,above ground fresh weight,above ground dry weight,root weight,root length,and stem diameter by 53.40%,67.50%,57.50%,73.50%,77.28%,and 9.40%,res

关 键 词：苯甲酸 酚酸降解菌 促生长 拮抗 番茄枯萎病 

分 类 号：S432.4[农业科学—植物病理学]