利用实时荧光定量PCR技术对耕地土壤中氨氧化细菌进行定量分析。
Quantification of ammonia-oxidizing bacteria in arable soil by real-time PCR.
作者信息
Hermansson A, Lindgren P E
机构信息
Department of Biology, Linköping University, S-58183 Linköping, Sweden.
出版信息
Appl Environ Microbiol. 2001 Feb;67(2):972-6. doi: 10.1128/AEM.67.2.972-976.2001.
Abstract
Real-time PCR was used to quantify populations of ammonia-oxidizing bacteria representing the beta subdivision of the class Proteobacteria in samples of arable soil, both nitrogen fertilized and unfertilized, from Mellby, Sweden. Primers and probes targeting a 16S ribosomal DNA region of the ammonia-oxidizing bacteria were designed and used. In the fertilized soil there were approximately 6.2 x 10(7) ammonia-oxidizing bacteria per g of soil, three times more than the number of bacteria in the unfertilized soil. The lytic efficiency of bead beating in these soils was investigated by using populations of free or loosely attached bacteria, bacteria tightly bound to particles, and bacteria in nonfractionated samples. The shapes of the curves generated in these tests showed that the concentration of template DNA released at various times remained constant after 10 to 100 s of bead beating.
摘要
采用实时定量聚合酶链反应（Real-time PCR）对瑞典梅利比（Mellby）地区施氮和未施氮耕地土壤样本中代表变形菌纲β亚类的氨氧化细菌数量进行定量分析。设计并使用了针对氨氧化细菌16S核糖体DNA区域的引物和探针。在施肥土壤中，每克土壤约有6.2×10⁷个氨氧化细菌，是未施肥土壤中细菌数量的三倍。通过对游离或松散附着的细菌、紧密结合在颗粒上的细菌以及未分级样本中的细菌群体进行研究，考察了珠磨法在这些土壤中的裂解效率。这些测试所生成曲线的形状表明，在珠磨10至100秒后，不同时间释放的模板DNA浓度保持恒定。
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