一株分泌型铁载体真菌分离鉴定及生物活性研究  

Isolation and Identification of a Secretory Siderophore Fungus,and Its Biological Activity

作　　者：邹雪峰 李铭刚[2] 包玲风 陈齐斌[1] 赵江源[2] 汪林 濮永瑜 郝大程[4] 张庆[5] 杨佩文[5] ZOU Xue-feng;LI Ming-gang;BAO Ling-feng;CHEN Qi-bin;ZHAO Jiang-yuan;WANG Lin;PU Yong-yu;HAO Da-cheng;ZHANG Qing;YANG Pei-wen(College of Plant Protection,Yunnan Agricultural University,Kunming 650201;Yunnan Institute of Microbiology,Yunnan University,Kunming 650091;Baoshan Tobacco Company of Yunnan Province,Baoshan 678000;School of Environment and Chemical Engineering,Dalian Jiaotong University,Dalian 116028;Institute of Agricultural Environment and Resources,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650201 [2]云南大学云南省微生物研究所,昆明650091 [3]云南省烟草公司保山市公司,保山678000 [4]大连交通大学环境化工学院/生物技术研究所,大连116028 [5]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》2022年第3期130-138,共9页Biotechnology Bulletin

基　　金：云南省重点领域科技计划绿色食品牌专项(202002AE320005);国家自然科学基金项目(31760019);云南省烟草公司科技计划重点项目(2020530000241021)。

摘　　要：从云南省国家自然保护区哀牢山原始森林土壤环境中分离1株分泌型铁载体真菌,并探讨了其对罹烟草青枯病土壤微生物生理生化功能的影响,为菌株的开发利用提供理论依据。稀释涂布法从土壤样品中分离纯化分泌型铁载体真菌菌株,形态学结合rDNA-ITS基因序列鉴定菌株分类学地位。铬天青S(chromeazurol S,CAS)液检测法结合全波段紫外光吸收法,判定铁载体化学结构类型,检测其活性。DNS比色法和Biolog-ECO分析法,分析菌株次生代谢产物对罹烟草青枯病土壤酶活性和微生物代谢的影响。结合形态学观察结果和序列对比分析鉴定该分泌型铁载体真菌为百岁兰曲霉(Aspergillus welwitschiae);菌株分泌型铁载体的化学结构类型为羧酸盐型(carboxylates),铁载体活性为78.79%。菌株次生代谢产物可显著提高土壤蔗糖酶和脲酶活性(P<0.05),分别提高13.7%和14.46%;菌株次生代谢产物亦可显著提高碳源利用能力:平均颜色变化率(average well color development,AWCD)和多样性指数(Shannon指数和Richness指数),分别提高86.15%、50.43%和66.67%。原始森林土壤环境贮藏铁载体微生物资源,利用菌株分泌型铁载体介导改善土壤微生物群落生理生化功能是实施土壤生态系统的优化管理的重要技术手段。A secretory siderophore producing fungus was isolated from the soil environment of Ailao Mountain primeval forest in Yunnan National Nature Reserve,and its effects on the physiological and biochemical functions of soil microorganisms infected with tobacco bacterial wilt were studied,which provides a theoretical basis for the development and utilization of the strain.Dilution coating method was used to isolate and purify the fungus producing the secretory siderophore from soil sample,the taxonomic status of the strain was identified by morphology and rDNA-ITS gene sequence.Chromazurol S(CAS)solution detection combined with full band ultraviolet absorption method was to determine the chemical structure type and activity of siderophore.DNS colorimetric method and Biolog-ECO analysis method were used to analyze the effects of strain secondary metabolites on the enzyme activity and microbial metabolism of tobacco bacterial wilt soil.The secretory siderophore-producing fungus was identified as Aspergillus welwitschiae based on morphological observations and sequence alignment analysis;the chemical structure type of the secretory siderophore of the strain was carboxylates,and its siderophore activity was 78.79%.The secondary metabolites of the strain significantly increased the activities of soil invertase and urease(P<0.05)by 13.7%and 14.46%respectively.The secondary metabolites of the strain also significantly improved the carbon source utilization capacity:Average color change rate(AWCD)and diversity index(Shannon index and richness index)increased by 86.15%,50.43%and 66.67%respectively.In conclusion,siderophore microbial resources are stored in primeval forest soil environment.The improvement of physiological and biochemical functions of the soil microbial community mediated by strain-secreted siderophore is an important technical means to optimize the management of the soil ecosystem.

关 键 词：原始森林土壤 分泌型铁载体真菌 烟草青枯病 土壤酶 Biolog-ECO 

分 类 号：S714.3[农业科学—林学]