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通过MCH-PCR继以巢式PCR预测土壤微生物矿化莠去津的潜力。

The potential of soil microorganisms to mineralize atrazine as predicted by MCH-PCR followed by nested PCR.

作者信息

Shapir N, Goux S, Mandelbaum R T, Pussemier L

机构信息

Institute of Soil, Water and Environmental Sciences, Volcani Center, Bet Dagan, Israel.

出版信息

Can J Microbiol. 2000 May;46(5):425-32. doi: 10.1139/w00-004.

Abstract

The potential of soil microorganisms to mineralize atrazine was studied in soil samples collected from fields with various histories of atrazine application. In contrast to many previous studies, which showed no atrazine mineralization activity, all the tested soils mineralized atrazine regardless of their atrazine application history. However, the delay before mineralization and the variation in the subsequent mineralization rate were in agreement with the initial copy number of the atrazine dechlorinaze gene, and the proliferation rate of the degraders. Soils from corn fields, which had up to 100 copies of the atzA gene per gram of soil, had a lag period of 4-5 days before atrazine mineralization started, and final mineralization percentages ranged from 40% to 54%. However, soils from fields that were never amended with atrazine had much longer lag periods (more than 17 days), which decreased after enrichment of the degrader population with high concentrations of atrazine for 15 days. Generally the mineralization rate and the atzA gene copy number increased after the enrichment period. The atrazine mineralization potential was measured by PCR of genes from the atrazine mineralization pathway. Magnetic capture hybridization was the most efficient of the two tested methods for purifying target DNA of PCR inhibitors, without reducing the copy number of the required fragment. Nested PCR proved to be the most effective method for predicting the exact potential of the soil to mineralize the pollutant even without enrichment of a small population with the target genes. This method can complement microcosm studies and eliminate futile efforts when the potential to mineralize the pollutant does not exist in the soil.

摘要

对从不同阿特拉津施用历史的农田采集的土壤样本中土壤微生物矿化阿特拉津的潜力进行了研究。与许多先前显示无阿特拉津矿化活性的研究不同,所有测试土壤均能矿化阿特拉津,无论其阿特拉津施用历史如何。然而,矿化前的延迟以及随后矿化速率的变化与阿特拉津脱氯酶基因的初始拷贝数以及降解菌的增殖速率一致。来自玉米田的土壤,每克土壤中atzA基因拷贝数高达100个,在阿特拉津矿化开始前有4 - 5天的滞后期,最终矿化百分比在40%至54%之间。然而,从未施用阿特拉津的农田土壤滞后期长得多(超过17天),在用高浓度阿特拉津富集降解菌群体15天后滞后期缩短。一般来说,富集期后矿化速率和atzA基因拷贝数增加。通过对阿特拉津矿化途径基因进行PCR来测定阿特拉津矿化潜力。磁捕获杂交是两种测试方法中最有效的用于纯化PCR抑制剂靶DNA的方法,且不会减少所需片段的拷贝数。巢式PCR被证明是预测土壤矿化污染物确切潜力的最有效方法,即使不通过靶基因富集少量群体也是如此。该方法可以补充微观世界研究,并在土壤中不存在矿化污染物潜力时消除徒劳的努力。

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