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采用实时定量 PCR 监测苯脲水解酶 A 和 B 编码基因评估敌草隆降解群落对敌草隆的响应。

Response of a diuron-degrading community to diuron exposure assessed by real-time quantitative PCR monitoring of phenylurea hydrolase A and B encoding genes.

机构信息

Irstea, UR MALY, 3 Bis Quai Chauveau-CP 220, 69336 Lyon, France.

出版信息

Appl Microbiol Biotechnol. 2013 Feb;97(4):1661-8. doi: 10.1007/s00253-012-4318-3. Epub 2012 Aug 2.

Abstract

A real-time quantitative PCR method was developed to detect and quantify phenlylurea hydrolase genes' (puhA and puhB) sequences from environmental DNA samples to assess diuron-degrading genetic potential in some soil and sediment microbial communities. In the soil communities, mineralization rates (determined with [ring-¹⁴C]-labeled diuron) were linked to diuron-degrading genetic potentials estimated from puhB number copies, which increased following repeated diuron treatments. In the sediment communities, mineralization potential did not depend solely on the quantity of puhB copies, underlining the need to assess gene expression. In the sediment samples, both puhB copy numbers and mineralization capacities were highly conditioned by whether or not diuron-treated soil was added. This points to transfers of degradative potential from soils to sediments. No puhA gene was detected in soil and sediment DNA extracts. Moreover, some sediments exhibited high diuron mineralization potential even though puhB genes were not detected, suggesting the existence of alternative diuron degradation pathways.

摘要

建立了一种实时定量 PCR 方法,用于从环境 DNA 样本中检测和定量苯脲水解酶基因(puhA 和 puhB)序列,以评估某些土壤和沉积物微生物群落中对苯嗪草酮的降解遗传潜力。在土壤群落中,矿化率(用[环-¹⁴C]-标记的苯嗪草酮测定)与从 puhB 拷贝数估计的苯嗪草酮降解遗传潜力相关,在重复苯嗪草酮处理后,其拷贝数增加。在沉积物群落中,矿化潜力不仅仅取决于 puhB 拷贝数的数量,这强调了需要评估基因表达。在沉积物样本中,puhB 拷贝数和矿化能力都受到是否添加苯嗪草酮处理土壤的强烈影响。这表明降解潜力从土壤转移到沉积物。在土壤和沉积物 DNA 提取物中均未检测到 puhA 基因。此外,一些沉积物表现出很高的苯嗪草酮矿化潜力,尽管未检测到 puhB 基因,这表明存在替代的苯嗪草酮降解途径。

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