3株多环芳烃高效降解菌株的分离鉴定及降解特性

doi:10.13287/j.1001-9332.202112.028

应用生态学报 ›› 2021, Vol. 32 ›› Issue (12): 4439-4446.doi: 10.13287/j.1001-9332.202112.028

3株多环芳烃高效降解菌株的分离鉴定及降解特性

闫双堆¹, 刘利军², 曹燕篆¹, 闫秋艳³, 董馨宇¹, 洪坚平^1*

¹山西农业大学资源环境学院, 山西晋中 030801;
²山西伊万泰克科技有限公司, 太原 030000;
³山西农业大学小麦研究所, 山西临汾 041000

收稿日期:2021-04-20 修回日期:2021-08-23 出版日期:2021-12-15 发布日期:2022-06-15
通讯作者: *E-mail: hongjp sx@163.com
作者简介:闫双堆, 女, 1976年生, 博士研究生。主要从事土壤污染与防治、固废资源化利用研究。E-mail: ysdllj@126.com
基金资助:
山西省科技攻关项目(20110311021)、山西省重点研发项目(201803D221003-2)和“降解多环芳烃优势菌筛选与菌群的组建项目(2010HX19)”资助

Isolation, identification, and degradation characteristics of three effective PAHs degradation strains

YAN Shuang-dui¹, LIU Li-jun², CAO Yan-zhuan¹, YAN Qiu-yan³, DONG Xin-yu¹, HONG Jian-ping^1*

¹College of Resources and Environment, Shanxi Agricultural University, Jinzhong 030801, Shanxi, China;
²Shanxi evantech Technology Co., Ltd, Taiyuan 030000, China;
³Institute of Wheat Research, Shanxi Agricultural University, Linfen 041000, Shanxi, China

Received:2021-04-20 Revised:2021-08-23 Online:2021-12-15 Published:2022-06-15
Contact: *E-mail: hongjp sx@163.com
Supported by:
Science and Technology Development Program of Shanxi Province (20110311021), Key Research and Development program of Shanxi Province (201803D221003-2) and “Screening and Forming of PAHs Degradation Dominant Bacteria (2010HX19)”

摘要/Abstract

摘要： 筛选分离降解多环芳烃(PAHs)的优势菌种对开展多环芳烃污染生态系统修复具有重要的现实意义。本研究以焦化厂周围受多环芳烃污染的土壤为菌源,经过富集培养驯化和平板分离,获得11株能降解多环芳烃的菌株。通过形态观察、生理生化特征及16S rRNA序列比对对菌株进行鉴定,筛选出3株PAHs高效降解菌,分别命名为DJ-3、DJ-8、DJ-10。经16S rRNA序列分析鉴定,DJ-3为假单胞菌属、DJ-8为克雷伯氏菌属、DJ-10为芽孢杆菌属。对菌株降解能力的研究表明,3株菌(DJ-3、DJ-8、DJ-10)培养7 d后对混合多环芳烃中菲(200 mg·L^-1)、芘(200 mg·L^-1)和萘(160 mg·L^-1)的降解率分别为48.9%～65.9%、38.9%～43.1%和57.6%～64.9%。3株菌对多环芳烃混合样品(1200 mg·L^-1)的降解率分别为49.1%、44.5%、53.9%,远高于其他8株筛选菌,为PAHs高效降解菌株。3种菌株两两之间和三者组合均无拮抗关系。研究结果将为构建高效的多环芳烃降解菌群、提高多环芳烃原位污染土壤的生物修复效果奠定基础。

关键词: 焦化厂, 多环芳烃, 降解菌株, 分离鉴定, 降解特性

Abstract: Isolating dominant strains for the degradation of polycyclic aromatic hydrocarbons (PAHs) is of great practical significance for the restoration of ecosystem polluted by PAHs. A total of 11 strains with capacity of degrading PAHs were obtained from soil polluted by PAHs around a coking plant, by enrichment culture, acclimation, and plate isolation. Three of them with effective PAH-degrading capability were identified and screened out by morphological observation, physiobiochemical characterization, and 16S rRNA gene sequencing, and respectively, named as DJ-3, DJ-8 and DJ-10. Based on the results of 16S rRNA gene sequencing, DJ-3, DJ-8, and DJ-10 were identified as Pseudomonas sp. Klebsiella sp., and Bacillus sp. The degradation rate of phenanthrene (200 mg·L^-1), pyrene (200 mg·L^-1), and naphthol (160 mg·L^-1) by three strains (DJ-3, DJ-8 and DJ-10) after seven-day incubation were 48.9%-65.9%, 38.9%-43.1%, and 57.6%-64.9%, respectively. The degradation rates of mixed PAHs sample (1200 mg·L^-1) by three strains were 49.1%, 44.5%, and 53.9%, which were significantly higher than other eight strains, indicating that they were highly effective in PAHs degradation. There was no antagonistic relationship among the three strains. This study would lay a foundation for building efficient PAHs degrading strains and improve the in situ bioremediation of PAHs contaminated soil.

Key words: coking plant, polycyclic aromatic hydrocarbons, degrading strains, isolation and identification, degradation characteristics

闫双堆, 刘利军, 曹燕篆, 闫秋艳, 董馨宇, 洪坚平. 3株多环芳烃高效降解菌株的分离鉴定及降解特性[J]. 应用生态学报, 2021, 32(12): 4439-4446.

YAN Shuang-dui, LIU Li-jun, CAO Yan-zhuan, YAN Qiu-yan, DONG Xin-yu, HONG Jian-ping. Isolation, identification, and degradation characteristics of three effective PAHs degradation strains[J]. Chinese Journal of Applied Ecology, 2021, 32(12): 4439-4446.

参考文献

[1] 马丹, 王永刚, 陈吉祥, 等. 1株高效菲降解不动杆菌的筛选、鉴定及性能研究. 微生物学杂志, 2018, 38(6): 15-23 [Ma D, Wang Y-G, Chen J-X, et al. Isolation, identification and properties of a high-efficient phenanthrene-degrading strain of Acinetobacter sp. Journal of Microbiology, 2018, 38(6): 15-23]
[2] 徐胜, 王慧, 陈玮, 等. 土壤中多环芳烃污染对植物生理生态的影响. 应用生态学报, 2013, 24(5): 1284-1290 [Xu S, Wang H, Chen W, et al. Effects of soil PAHs pollution on plant ecophysiology. Chinese Journal of Applied Ecology, 2013, 24(5): 1284-1290]
[3] 张娟琴, 陈思尹, 唐卫红, 等. 高效多环芳烃降解菌的筛选、鉴定及降解特性分析. 上海农业学报, 2020, 36(1): 98-104 [Zhang J-Q, Chen S-Y, Tang W-H, et al. Screening, identification and degradative characteristics of PAHs degrading bacteria. Acta Agriculturae Shanghai, 2020, 36 (1): 98-104]
[4] 高秀荣, 林姗娜, 孙安琪, 等. 一株多环芳烃降解菌的筛选及其降解特性. 环境工程学报, 2020, 14(6): 1668-1678 [Gao X-R, Lin S-N, Sun A-Q, et al. Screening and degradation characteristics of a PAHs-degrading bacteria. Chinese Journal of Environmental Engineering, 2020, 14 (6): 1668-1678]
[5] 李荣峰, 赵碧川, 陈丽君, 等. 多环芳烃降解菌的分离、鉴定及驯化研究. 安徽农学通报, 2017, 23(18): 13-15, 42 [Li R-F, Zhao B-C, Chen L-J, et al. Isolation and identification and domestication of PAHs degrading bacteria. Anhui Agricultural Science Bulletin, 2017, 23(18): 13-15, 42]
[6] 胡凤钗, 苏振成, 孙健, 等. 高效芘降解菌N12的分离鉴定与降解特性. 应用生态学报, 2011, 22(6): 1566-1572 [Hu F-C, Su Z-C, Sun J, et al. Isolation and identification of a highly efficient pyrene-degrading Mycobacterium sp. strain N12. Chinese Journal of Applied Ecology, 2011, 22(6): 1566-1572]
[7] 毛健, 骆永明, 滕应, 等. 一株高分子量多环芳烃降解菌的筛选、鉴定及降解特性研究. 微生物学通报, 2008, 35(7): 1011-1015 [Mao J, Luo Y-M, Teng Y, et al. Isolation and characterization of, a high-molecular-weight (HMW) PAHs degrading bacterial strain. Microbiology China, 2008, 35 (7): 1011-1015]
[8] 任静, 沈佳敏, 张磊, 等. 生物炭固定化多环芳烃高效降解菌剂的制备及稳定性. 环境科学学报, 2020, 40(12): 4517-4523 [Ren J, Shen J-M, Zhang L, et al. Preparation and stability of bacteria for the immobilization of polycyclic aromatic hydrocarbons degradating-bacteria. Acta Scientiae Circumstantiae, 2020, 40(12): 4517-4523]
[9] 曹阳, 宋立超, 钮旭光, 等. 焦化厂地肤根内解芘细菌的筛选及促生潜力. 应用生态学报, 2019, 30(6): 2056-2062 [Cao Y, Song L-C, Niu X-G, et al. Screening of pyrene degradation bacteria strain from the Kochia scoparia roots and its potential for promoting growth. Chinese Journal of Applied Ecology,2019, 30(6):2056-2062]
[10] 刘楠楠, 陈鹏, 朱淑贞, 等. 辽河和太湖沉积物中PAHs和OCPs的分布特征及风险评估. 中国环境科学, 2011, 31(2): 293-300 [Liu N-N, Chen P, Zhu S-Z, et al. Distribution characteristics of PAHs and OCPs in sediments of Liaohe River and Taihu Lake and their risk evaluation based on quality criteria. China Environmental Science, 2011, 31(2): 293-300]
[11] 卢晓霞, 吴淑可, 侯珍, 等. 焦化厂污染土壤中多环芳烃降解菌的分离及降解特性. 应用与环境生物学报, 2011, 17(5): 722-726 [Lu X-X, Wu S-K, Hou Z, et al. Isolation and degrading properties of PAHs-degrading bacteria from the contaminated soil of a coking plant. Chinese Journal of Applied and Environmental Biology, 2011, 17(5): 722-726]
[12] 肖盟, 尹向阳, 马红蕾, 等. 多环芳烃降解菌的筛选及其降解性能的强化. 煤炭科学技术, 2018, 46(9): 75-80 [Xiao M, Yin X-Y, Ma H-L, et al. Screening and degrading capacity improvement of polycyclic aromatic hydrocarbons degrading bacterium. Coal Science and Technology, 2018, 46(9): 75-80]
[13] 王新廷, 王芸, 赵传富, 等. 多环芳烃降解菌的筛选、鉴定及其特性. 曲阜师范大学学报: 自然科学版, 2019, 45(3): 92-96 [Wang X-T, Wang Y, Zhao C-F, et al. Isolation, ientification and caracterization of plycyclic aomatic hdrocarbon dgrading bcteria. Journal of Qufu Normal University: Natural Science, 2019,45 (3): 92-96]
[14] Wilson SC, Jones KC. Bioremediation of soil contaminated with polynuclear aromatic hydrocarbons (PAHs): A review. Environmental Pollution, 1993, 81: 229-249
[15] 田晶, 徐小琳, 康彦顺, 等. 广谱性多环芳烃降解真菌Aspergillus flavus AD-X-1 的筛选及其性能研究. 生物技术通报, 2018, 34(8): 115-122 [Tian J, Xu X-L, Kang Y-S, et al. Screening and characterization of a broad spectrum fungus degrading polycyclic-aromatic hydrocarbons: Aspergillus flavus AD-X-1. Biotechnology Bulletin, 2018, 34(8): 115-122]
[16] 杨轩, 张威, 李师翁, 等. 多环芳烃降解菌的分离鉴定及其生理特性研究. 环境科学学报, 2012, 32(5): 1033-1040 [Yang X, Zhang W, Li S-W, et al. Study on isolation, identification and physiological characteristics of PAHs-degrading bacteria. Acta Scientiae Circumstantiae, 2012, 32(5): 1033-1040]
[17] 高野萌, 杨雪莲, 李凤梅, 等. 高环多环芳烃降解菌的筛选及其降解特性. 生态学杂志, 2016, 35(6):1539-1546 [Gao Y-M, Yang X-L, Li F-M, et al. Isolation of heavy PAH-degrading bacteria and their characteristics of degradation. Chinese Journal of Ecology, 2016, 35(6): 1539-1546]
[18] 范瑞娟, 刘雅琴, 张琇. 嗜盐碱高环PAHs降解菌的分离及其降解特性研究. 农业环境科学学报, 2019, 38(6): 1280-1287 [Fan R-J, Liu Y-Q, Zhang X. Isolation and degradation characterization of haloalkaliphilic high molecular-weight polycyclic aromatic hydro-carbon-degrading bacteria. Journal of Agro-Environment Science, 2019, 38(6): 1280-1287]
[19] 黄兴如, 张彩文, 张瑞杰, 等. 多环芳烃降解菌的筛选、鉴定及降解特性.微生物学通报, 2016, 43(5): 965-973 [Huang X-R, Zhang C-W, Zhang R-J, et al. Isolation, identification and degradating properties of PAHs-degrading bacteria. Microbiology China, 2016, 43(5): 965-973]
[20] 刘利军. 太原市小店污灌区农田安全利用技术研究. 博士论文. 太谷: 山西农业大学, 2013 [Liu L-J. Research on Farmland Safe Utilization in Xiaodian Sewage Irrigation Area, Taiyuan. PhD Thesis. Taigu: Shanxi Agricultural University, 2013]
[21] 肖琳, 杨柳燕, 尹大强, 等. 环境微生物实验技术. 北京: 中国环境科学出版社, 2004: 253-261 [Xiao L, Yang L-Y, Yin D-Q, et al. Protocols of Environmental Microbiology. Beijing: China Environmental Science Press, 2004: 253-261]
[22] Buchanan RE, Gibbons NE. 中国科学院微生物研究所《伯杰细菌鉴定手册》翻译组, 译. 伯杰氏细菌鉴定手册. 第八版. 北京: 科学出版社, 1984: 729-735 [Buchanan RE, Gibbons NE, eds. Trans: Translation team of Berger’s Manual of bacterial identification, Institute of Microbiology, Chinese Academy of Sciences, Bergey’s Manual of Determinative Bacteriology. 8th Ed. Beijing: Science Press, 1984: 729-735]
[23] 李振高, 骆永明, 滕应. 土壤与环境微生物研究法. 北京: 科学出版社, 2008 [Li Z-G, Luo Y-M, Teng Y. Research Methods of Soil and Environmental Microbiology. Beijing: Science Press, 2008]
[24] 乔志伟. 石灰性土壤溶磷细菌的筛选鉴定及在复垦土壤上的应用. 博士论文. 太谷: 山西农业大学, 2014 [Qiao Z-W. Screening and Identification of Phosphate Solubilizing Bacteria in Calcareous Soil and the Application in Coal Mining Reclaimed Soil. PhD Thesis. Taigu: Shanxi Agricultural University, 2014]
[25] Geiselbrecht AD, Herwig RP, Deming JW, et al. Enumeration and phylogenetic analysis of polycyclic aromatic hydrocarbon-degrading marine bacteria from Puget sound sediments. Applied and Environmental Microbiology, 1996, 62: 3344-3349
[26] Canet R, Birnstingl JG, Malcolm DG, et al. Biodegradation of polycyclic aromatic hydrocarbons (PHAs) by native microflora and combinations of white-rot fungi in a coal-tar contaminated soil. Bioresource Technolgy, 2001, 76: 113-117
[27] 孟祥帅, 陈鸿汉, 郑从奇, 等. 焦化厂不同污染源作用下土壤PAHs污染特征. 中国环境科学, 2020, 40(11): 4857-4864 [Meng X-S, Chen H-H, Zheng C-Q, et al. Pollution characteristics of PAHs in soil at an abandoned coking plant affected by different sources. China Environmental Science, 2020, 40(11): 4857-4864]
[28] 潘峰, 耿秋娟, 楚红杰, 等. 石油污染土壤中多环芳烃分析及生态风险评价. 生态与农村环境学报, 2011, 27(5): 42-47 [Pan F, Geng Q-J, Chu H-J, et al. Analysis of polycyclic aromatic hydrocarbons in petroleum contaminated soil and its ecological risk assessment. Journal of Ecology and Rural Environment, 2011, 27(5): 42-47]
[29] 冯嫣, 吕永龙, 焦文涛, 等. 北京市某废弃焦化厂不同车间土壤中多环芳烃(PHAs)的分布特征及风险评价. 生态毒理学报, 2009, 4(3): 399-407 [Feng Y, Lv Y-L, Jiao W-T, et al. Distribution and risk of polycyclic aromatic hydrocarbons in soil from different workshops of an abandoned coking plant in Beijing. Asian Journal of Ecotoxicology, 2009, 4(3): 399-407]
[30] Chang JS, Cha DK, Radosevich M, et al. Effects of biosurfactant-producing bacteria on biodegradation and transport of phenathrene in subsurface soil. Journal of Environmental Science and Health, Part A: Toxic/Hazardous Substances and Evironmental Engineering, 2015, 50: 611-616]
[31] 李凤梅, 郭书海, 张灿灿, 等. 多环芳烃降解菌的筛选及其在焦化场地污染土壤修复中的应用. 环境污染与防治, 2016, 38(4): 1-5 [Li F-M, Guo S-H, Zhang C-C, et al. Isolation of PAHs degrading bacteria and its application to mediation of pollution soil in coking site. Environment Pollution & Control, 2016, 38(4): 1-5]
[32] 杨晓磊, 陆贻通, 曹林奎. 多环芳烃荧蒽降解菌的筛选鉴定及降解特性研究. 科技通报, 2007, 23(1): 46-51 [Yang X-L, Lu Y-T, Cao L-K. Studys on isolation identification of a fluoranthene-degrading strains and its degradation characteristics. Bulletin of Science and Technology, 2007, 23(1): 46-51]
[33] 王春明, 李大平, 刘世贵. 4株多环芳烃降解菌的分离与鉴定. 应用与环境生物学报, 2007, 13(4): 546-550 [Wang C-M, Li D-P, Liu S-G. Screening and identification of four PAHs-biodegrading strains. Chinese Journal of Applied and Environmental Biology, 2007, 13(4): 546-550]
[34] 王春明, 李大平, 王春莲. 微杆菌3-28对萘、菲、蒽、芘的降解. 应用与环境生物学报, 2009, 15(3): 361-366 [Wang C-M, Li D-P, Wang C-L. Biodegradation of naphthalene, phenanathrene, anthracene and pyrene by Microbacterium sp. 3-28. Chinese Journal of Applied and Environmental Biology, 2009, 15(3): 361-366]
[35] 王佩, 蒋鹏, 张华, 等. 焦化厂土壤和地下水中PAHs 分布特征及其污染过程. 环境科学研究, 2015, 28(5): 752-759 [Wang P, Jiang P, Zhang H, et al. Distribution characteristics and pollution process of PAHs in soil and groundwater at a coking plant site. Research of Environmental Sciences, 2015, 28(5): 752-759]
[36] 倪妮, 宋洋, 王芳, 等. 多环芳烃污染土壤生物联合强化修复研究进展. 土壤学报, 2016, 53(3): 561-571 [Ni N, Song Y, Wang F, et al. A review of researches on intensified bio-remediation of polycyclic aromatic hydrocarbons contaminated soils. Acta Pedologica Sinica, 2016, 53(3): 561-571]

3株多环芳烃高效降解菌株的分离鉴定及降解特性

Isolation, identification, and degradation characteristics of three effective PAHs degradation strains

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[10]	王菲;苏振成;杨辉;李晓军;杨官品;董殿波. 土壤中多环芳烃的微生物降解及土壤细菌种群多样性 [J]. 应用生态学报, 2009, 20(12): 3020-3026.
[11]	刘泓¹;叶媛蓓¹;崔波²;郑荔敏¹;黄炎和¹;王宗华². 多环芳烃荧蒽诱导拟南芥氧化胁迫 [J]. 应用生态学报, 2008, 19(02): 413-418 .
[12]	郭伟;何孟常;杨志峰;林春野;全向春;王浩正. 大辽河水系表层水中多环芳烃的污染特征 [J]. 应用生态学报, 2007, 18(07): 1534-1538 .
[13]	任瑞霞^1,2;张颖¹;李慧¹;王荣¹;徐慧¹;陈冠雄¹. 石油污染土壤水改旱田后污染物组份及微生物群落结构变化 [J]. 应用生态学报, 2007, 18(05): 1107-1112 .
[14]	苏丹^1,2;李培军¹;鞠京丽¹;许华夏¹. 非流体介质中多环芳烃污染的微生物固定化修复技术 [J]. 应用生态学报, 2006, 17(08): 1530-1534 .
[15]	王曙光¹;冯兆忠²;王效科². 环境污染物对丛枝菌根(AM)形成及功能的影响 [J]. 应用生态学报, 2006, 17(07): 1321-1325 .