Department of Soil Science, Kasetsart University, Bangkok, Thailand.
J Microbiol Biotechnol. 2010 Jan;20(1):169-78.
The microbial community structure in Thailand soils contaminated with low and high levels of arsenic was determined by denaturing gradient gel electrophoresis (DGGE). Band pattern analysis indicated that the bacterial community was not significantly different in the two soils. Phylogenetic analysis obtained by excising and sequencing six bands indicated that the soils were dominated by Arthobacter koreensis and proteobacteria. Two hundred and sixty-two bacterial isolates were obtained from arsenic contaminated soils. The majority of the As resistant isolates were gram-negative bacteria. MIC studies indicated that all of the tested bacteria had greater resistance to arsenate than arsenite. Some strains were capable of growing in medium containing up to 1,500 mg/l arsenite and arsenate. Correlations analysis of resistance patterns of arsenite resistance indicated that the isolated bacteria could be categorized into 13 groups, with a maximum similarity value of 100%. All strains were also evaluated for resistance to eight antibiotics. The antibiotic resistance patterns divided the strains into 100 unique groups, indicating that the strains were very diverse. Isolates from each antibiotic resistance group were characterized in more detail by using the repetitive extragenic palindromic-PCR (rep-PCR) DNA fingerprinting technique with ERIC primers. PCR products were analyzed by agarose gel electrophoresis. The genetic relatedness of 100 bacterial fingerprints, determined by using Pearson product moment similarity coefficient, showed that the isolates could be divided into four clusters, with similarity values ranging from 5-99%. While many isolates were genetically diverse, others were clonal in nature Additionally, the arsenic-resistant isolates were examined for the presence of arsenic resistance (ars) genes by using PCR, and 30% of the isolates were found to carry an arsenate reductase encoded by the arsC gene.
采用变性梯度凝胶电泳(DGGE)技术研究了泰国低砷和高砷污染土壤中的微生物群落结构。条带图谱分析表明,两种土壤中的细菌群落无显著差异。通过切胶测序获得的系统发育分析表明,土壤中主要的细菌类群为节杆菌(Arthobacter koreensis)和变形菌(Proteobacteria)。从砷污染土壤中分离得到 262 株细菌。大部分砷抗性菌株为革兰氏阴性菌。MIC 研究表明,所有测试的细菌对砷酸盐的抗性均大于亚砷酸盐。有些菌株能够在含有高达 1500mg/L 亚砷酸盐和砷酸盐的培养基中生长。砷酸盐抗性的抗性模式相关性分析表明,分离出的细菌可分为 13 组,最大相似性值为 100%。所有菌株还进行了对 8 种抗生素的抗性评估。抗生素抗性模式将菌株分为 100 个独特的组,表明菌株非常多样化。用重复回文外扩增 PCR(rep-PCR)ERIC 引物 DNA 指纹图谱技术对每个抗生素抗性组的菌株进行了更详细的特征描述。PCR 产物用琼脂糖凝胶电泳分析。通过使用 Pearson 乘积矩相似系数,对 100 个细菌指纹图谱的遗传相关性进行分析,结果表明,这些分离物可分为 4 个聚类,相似度值在 5-99%之间。虽然许多分离物在遗传上具有多样性,但其他分离物在本质上是克隆的。此外,通过 PCR 检测砷抗性分离物是否存在砷抗性(ars)基因,发现 30%的分离物携带由 arsC 基因编码的砷酸盐还原酶。