Bazhanov Dmitry P, Li Chengyun, Li Hongmei, Li Jishun, Zhang Xinjian, Chen Xiangfeng, Yang Hetong
Key Laboratory for Applied Microbiology of Shandong Province, Ecology Institute (Biotechnology Center) of Shandong Academy of Sciences, Jinan, Shandong Province, People's Republic of China.
Biology Institute of Shandong Academy of Sciences, Jinan, Shandong Province, People's Republic of China.
BMC Microbiol. 2016 Nov 8;16(1):265. doi: 10.1186/s12866-016-0868-3.
Soil populations of bacteria rapidly degrading atrazine are critical to the environmental fate of the herbicide. An enrichment bias from the routine isolation procedure prevents studying the diversity of atrazine degraders. In the present work, we analyzed the occurrence, diversity and community structure of soil atrazine-degrading bacteria based on their direct isolation.
Atrazine-degrading bacteria were isolated by direct plating on a specially developed SM agar. The atrazine degradation genes trzN and atzABC were detected by multiplex PCR. The diversity of atrazine degraders was characterized by enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) genotyping followed by 16S rRNA gene phylogenetic analysis. The occurrence of atrazine-degrading bacteria was also assessed by conventional PCR targeting trzN and atzABC in soil DNA.
A total of 116 atrazine-degrading isolates were recovered from bulk and rhizosphere soils sampled near an atrazine factory and from geographically distant maize fields. Fifteen genotypes were distinguished among 56 industrial isolates, with 13 of them representing eight phylogenetic groups of the genus Arthrobacter. The remaining two were closely related to Pseudomonas alcaliphila and Gulosibacter molinativorax and constituted major components of the atrazine-degrading community in the most heavily contaminated industrial plantless soil. All isolates from the adjacent sites inhabited by cogon grass or common reed were various Arthrobacter spp. with a strong prevalence of A. aurescens group. Only three genotypes were distinguished among 60 agricultural strains. Genetically similar Arthrobacter ureafaciens bacteria which occurred as minor inhabitants of cogon grass roots in the industrial soil were ubiquitous and predominant atrazine degraders in the maize rhizosphere. The other two genotypes represented two distant Nocardioides spp. that were specific to their geographic origins.
Direct plating on SM agar enabled rapid isolation of atrazine-degrading bacteria and analysis of their natural diversity in soil. The results obtained provided evidence that contaminated soils harbored communities of genetically distinct bacteria capable of individually degrading and utilizing atrazine. The community structures of culturable atrazine degraders were habitat-specific. Bacteria belonging to the genus Arthrobacter were the predominant degraders of atrazine in the plant rhizosphere.
能快速降解莠去津的土壤细菌群体对该除草剂的环境归宿至关重要。常规分离程序产生的富集偏差妨碍了对莠去津降解菌多样性的研究。在本研究中,我们基于直接分离分析了土壤中莠去津降解菌的存在情况、多样性和群落结构。
通过直接接种在专门研制的SM琼脂上分离莠去津降解菌。采用多重PCR检测莠去津降解基因trzN和atzABC。通过肠杆菌重复基因间共有序列PCR(ERIC-PCR)基因分型,随后进行16S rRNA基因系统发育分析,对莠去津降解菌的多样性进行表征。还通过针对土壤DNA中trzN和atzABC的常规PCR评估莠去津降解菌的存在情况。
从一家莠去津工厂附近采集的大量土壤和根际土壤以及地理上相距遥远的玉米田中,共分离出116株莠去津降解菌。在56株工业菌株中区分出15种基因型,其中13种代表节杆菌属的8个系统发育组。其余两种与嗜碱假单胞菌和食木古洛菌密切相关,是污染最严重的无植物工业土壤中莠去津降解群落的主要组成部分。来自相邻芒草或芦苇生长地的所有分离株均为各种节杆菌属物种,其中金色节杆菌组占优势。在60株农业菌株中仅区分出3种基因型。在工业土壤中作为芒草根际少量定殖菌出现的基因相似的解脲节杆菌,在玉米根际是普遍存在且占主导地位的莠去津降解菌。另外两种基因型代表两种与其地理来源相关的遥远的诺卡氏菌属物种。
直接接种在SM琼脂上能够快速分离莠去津降解菌,并分析其在土壤中的自然多样性。获得的结果提供了证据,表明受污染土壤中存在能够单独降解和利用莠去津的遗传上不同的细菌群落。可培养的莠去津降解菌的群落结构具有生境特异性。节杆菌属细菌是植物根际莠去津的主要降解菌。