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对南极洲和巴西土壤中本土微生物碳氢化合物降解基因的调查。

A survey of indigenous microbial hydrocarbon degradation genes in soils from Antarctica and Brazil.

作者信息

Luz A P, Pellizari V H, Whyte L G, Greer C W

机构信息

Biomedical Sciences Institute, Department of Microbiology, University of São Paulo, Brazil.

出版信息

Can J Microbiol. 2004 May;50(5):323-33. doi: 10.1139/w04-008.

Abstract

Total community DNA from 29 noncontaminated soils and soils impacted by petroleum hydrocarbons and chloro-organics from Antarctica and Brazil were screened for the presence of nine catabolic genes, encoding alkane monooxygenase or aromatic dioxygenases, from known bacterial biodegradation pathways. Specific primers and probes targeting alkane monooxygenase genes were derived from Pseudomonas putida ATCC 29347 (Pp alkB), Rhodococcus sp. strain Q15 (Rh alkB1, Rh alkB2), and Acinetobacter sp. ADP-1 (Ac alkM). In addition, primers and probes detecting aromatic dioxygenase genes were derived from P. putida ATCC 17484 (ndoB), P. putida F1 (todC1), P. putida ATCC 33015 (xylE and cat23), and P. pseudoalcaligenes KF707 (bphA). The primers and probes were used to analyze total community DNA extracts by using PCR and hybridization analysis. All the catabolic genes, except the Ac alkM, were detected in contaminated and control soils from both geographic regions, with a higher frequency in the Antarctic soils. The alkane monooxygenase genes, Rh alkB1 and Rh alkB2, were the most frequently detected alk genes in both regions, while Pp alkB was not detected in Brazil soils. Genes encoding the aromatic dioxygenases toluene dioxygenase (todC1) and biphenyl dioxygenase (bphA) were the most frequently detected in Antarctica, and todC1 and catechol-2,3-dioxygenase (cat23) were the most frequent in Brazil soils. Hybridization analysis confirmed the PCR results, indicating that the probes used had a high degree of homology to the genes detected in the soil extracts and were effective in detecting biodegradative potential in the indigenous microbial population.

摘要

对来自南极洲和巴西的29份未受污染土壤以及受石油烃和氯代有机物污染的土壤中的总群落DNA进行筛选,以检测已知细菌生物降解途径中9种编码烷烃单加氧酶或芳香族双加氧酶的分解代谢基因的存在情况。靶向烷烃单加氧酶基因的特异性引物和探针源自恶臭假单胞菌ATCC 29347(Pp alkB)、红球菌属菌株Q15(Rh alkB1、Rh alkB2)和不动杆菌属ADP - 1(Ac alkM)。此外,检测芳香族双加氧酶基因的引物和探针源自恶臭假单胞菌ATCC 17484(ndoB)、恶臭假单胞菌F1(todC1)、恶臭假单胞菌ATCC 33015(xylE和cat23)以及假产碱假单胞菌KF707(bphA)。这些引物和探针用于通过PCR和杂交分析来分析总群落DNA提取物。除Ac alkM外,所有分解代谢基因在两个地理区域的污染土壤和对照土壤中均被检测到,在南极土壤中的出现频率更高。烷烃单加氧酶基因Rh alkB1和Rh alkB2是两个区域中最常检测到的烷基因,而Pp alkB在巴西土壤中未被检测到。编码芳香族双加氧酶甲苯双加氧酶(todC1)和联苯双加氧酶(bphA)的基因在南极洲最常被检测到,而todC1和儿茶酚 - 2,3 - 双加氧酶(cat23)在巴西土壤中最常见。杂交分析证实了PCR结果,表明所使用的探针与土壤提取物中检测到的基因具有高度同源性,并且能够有效地检测本地微生物群落中的生物降解潜力。

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