Pseudomonas stutzeri nitrite reductase gene abundance in environmental samples measured by real-time PCR
Pseudomonas stutzeri nitrite reductase gene abundance in environmental samples measured by real-time PCR
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DOI：
10.1128/aem.67.2.760-768.2001
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发表时间：
2001-02-01
影响因子：
4.4
通讯作者：
Tiedje, JM
中科院分区：
文献类型：
--
作者：
Grüntzig, V;Nold, SC;Tiedje, JM
我们使用实时PCR定量的亚硝酸盐还原酶基因（nirS），一个功能基因的地球化学意义。该测定法在体外进行了测试，并适用于环境样品。选择的引物探针组对nirS序列具有特异性，该序列大致对应于斯氏假单胞菌物种。该测定在1至10（6）个基因拷贝之间呈线性（r（2）= 0.999）。在低基因浓度的变异性不允许检测基因拷贝数在小于100个拷贝的两倍差异。DNA掺入和细胞添加实验给出了预测结果，表明该测定提供了环境样品中斯氏疟原虫nirS丰度的准确测量。虽然P. stutzeri丰度在湖泊沉积物和地下水样品中很高，但我们在普吉特湾（华盛顿州）的海洋沉积物样品中检测到该物种的丰度很低或没有。和来自华盛顿海洋边缘的。这些结果表明，斯氏原杆菌可能不是一种优势海洋脱氮菌。
We used real-time PCR to quantify the denitrifying nitrite reductase gene (nirS), a functional gene of biogeochemical significance. The assay was tested in vitro and applied to environmental samples. The primer-probe set selected was specific for nirS sequences that corresponded approximately to the Pseudomonas stutzeri species. The assay was linear from 1 to 10(6) gene copies (r(2) = 0.999). Variability at low gene concentrations did not allow detection of twofold differences in gene copy number at less than 100 copies. DNA spiking and cell-addition experiments gave predicted results, suggesting that this assay provides an accurate measure of P. stutzeri nirS abundance in environmental samples. Although P. stutzeri abundance was high in lake sediment and groundwater samples, we detected low or no abundance of this species in marine sediment samples from Puget Sound (Wash.) and from the Washington ocean margin. These results suggest that P. stutzeri may not be a dominant marine denitrifier.