菌株Trametes hirsuta zlh237发酵液对污染土壤中多环芳烃的降解及群落结构分析  

Biodegradation and community structure analysis of polycyclic aromatic hydrocarbon(PAH)in contaminated soil by Trametes hirsuta zlh237 fermentation liquid

作　　者：甄静[1] 李冠杰[1] 杜志敏[1] 王继雯[1] 杨文玲[1] 巩涛[1] 赵俊杰[1] ZHEN Jing;LI Guan-jie;DU Zhi-min;WANG Ji-wen;YANGWen-ling;GONG Tao;ZHAO Jun-jie(Institute of Biology Co.,Ltd.,Henan Academy of Sciences/Key Laboratory of Microbial Engineering of Henan,Zhengzhou 450008,China)

机构地区：[1]河南省科学院生物研究所有限责任公司/河南省微生物工程重点实验室

出　　处：《南方农业学报》2020年第1期72-79,共8页Journal of Southern Agriculture

基　　金：河南省科技攻关项目(182102311032);河南省科学院科研开发项目(18ZX05003);河南省科学院基本科研业务费项目(190605020,190605019)~~

摘　　要：【目的】利用中国科学院微生物研究所真菌学国家重点实验室分离获得的菌株Trametes hirsuta zlh237,研究生物强化(接种菌株T.hirsuta zlh237发酵液)对污染土壤中3种多环芳烃(PAHs)[Phenanthrene、Pyrene和Benzo[a]pyrene(BaP)]的降解效果,为该菌株在PAHs污染土壤中的应用提供科学依据。【方法】采用高效液相色谱(HPLC)检测7个处理土壤(接种菌株T.hirsuta zlh237发酵液的Phenanthrene、Pyrene和BaP污染土壤分别标记为PheBA、PyrBA和BaPBA,接种灭菌发酵液的污染土壤分别标记为Phe、Pyr和BaP,原始土壤标记为CK)中3种PAHs含量,利用ABTS法检测土壤中漆酶活性,并运用高通量测序技术分析修复后土壤中细菌群落结构的变化。【结果】菌株T.hirsuta zlh237在3种PAHs污染土壤中均能生长,接种菌株发酵液15 d后,3种PAHs均有一定降解,其中BaP降解效果最佳,降解率达33.99%;且菌株T.hirsuta zlh237在3种污染土壤中均能分泌漆酶。高通量测序Alpha多样性指数分析结果表明,污染土壤接种菌株T.hirsuta zlh237发酵液后,Chao1指数和Shannon指数显著增加(P<0.05),不同处理土壤样品的Chao1指数和Shannon指数排序均为:PheBA>PyrBA>BaPBA>CK>Phe>Pyr>BaP。Beta多样性的主成分分析(PCA)结果表明,接种菌株T.hirsuta zlh237发酵液能改变污染土壤细菌群落结构组成;在门分类水平上,污染土壤样品中变形菌门(Proteobacteria)为优势门;在属分类水平上,接种菌株T.hirsuta zlh237发酵液的污染土壤样品中鞘氨醇单胞菌属(Sphingomonas)为优势菌属,接种灭菌发酵液的污染土壤Phe和Pyr样品中苯基杆菌属(Phenylobacterium)为优势菌属,而BaP样品中假单胞菌属(Pseudomonas)为优势菌属。【结论】菌株T.hirsuta zlh237发酵液在PAHs污染土壤中能分泌漆酶,对3种PAHs均有一定的降解效果,且能改变污染土壤细菌群落结构组成,在一定程度上对PAHs污染土壤有较好的修复效果。【Objective】Trametes hirsuta zlh237 isolated by State Key Laboratory of Mycology,Institute of microbiology,Chinese Academy of Sciences was used.Bioaugmentation by inoculating T.hirsuta zlh237 fermentation liquid was used to study its biodegradation effect on three kinds of polycyclic aromatic hydrocarbon(PAHs)[Phenanthrene,Pyrene and Benzo[a]pyrene(BaP)].That provided a scientific basis for its application in PAHs contaminated soil.【Method】The concentrations of the respective PAHs in the seven treated soil were determined by high performance liquid chromatography(HPLC)(The Phenanthrene,Pyrene and Bap contaminated soils incubated with fungal fermentation liquid were respectively labeled PheBA,PyrBA and BaPBA.The contaminated soils incubated with sterilized fungal fermentation liquid were respectively labeled Phe,Pyr and BaP.The original soil sample with sterilized water was labeled CK).Laccase activity was spectrophotometrically determined with ABTS.Bacterial community structures in the three PAHs-contaminated soils were analyzed by illumina MiSeq high-throughput sequencing.【Result】The results showed T.hirsuta zlh237 could remain active in the PAHs-environment.T.hirsuta showed the ability to degrade three PAHs in the soil,and had best degradation effect on BaP after 15 days with the degradation rate of 33.99%,and T.hirsuta zlh237 produced laccase in three PAHs contaminated soils.Moreover,the result of illumina MiSeq high-throughput sequencing alpha diversity analysis showed that the PAHs-contaminated soil inoculating T.hirsuta fermentation liquid showed significant increases(P<0.05)in the Chao1 index and Shannon index with the descending order of PheBA>PyrBA>BaPBA>CK>Phe>Pyr>BaP.Principal component analysis(PCA)of beta diversity implied that the microbiological structures of the PAHs-contaminated soil were restored by inoculating of T.hirsuta.At the phylum level,Proteobacteria was dominant phylum in the PAHs-contaminated soil.At the genus level,Sphingomonas was dominant genus in the PAHs-contaminated so

关 键 词：菌株Trametes hirsuta zlh237 发酵液 生物强化 多环芳烃 降解 高通量测序 群落结构 

分 类 号：S154.36[农业科学—土壤学]