Departamento de Física Aplicada, Escuela de Ingenierías Agrarias, Universidad de Extremadura, Avda, de Adolfo Suárez S/n, 06007 Badajoz, Spain.
Expresión Gráfica, Escuela de Ingenierías Industriales, Campus Universitario, Avda de Elvas sn, 06006 Badajoz, Spain.
J Microbiol Methods. 2022 Aug;199:106529. doi: 10.1016/j.mimet.2022.106529. Epub 2022 Jun 27.
Anaerobic digestion is a growing technology to manage organic waste and produce bioenergy. To promote this technology, it is essential to know, at the molecular level, the dynamics of microbial communities, specifically the methanogenic community. In the present study, three primer pairs were selected from seven primer pairs which were designed and tested with different concentrations and conditions to detect Methanosarcina, Methanoculleus and Methanobacterium by real-time PCR based on the SYBR Green System. The functionality of the developed methods was demonstrated by the high linear relationship of the standard curves, and the specificity of each primer was empirically verified by testing DNA isolated from methane-producing and non-producing strains. These assays also exhibited good repeatability and reproducibility, which indicates the robustness of the methods. The described primers were successfully used to investigate the methanogenic communities of 10 samples from an anaerobic co-digestion. The genus Methanosarcina was the dominant methanogenic group.
厌氧消化是一种处理有机废物并生产生物能源的新兴技术。为了推广这项技术，必须在分子水平上了解微生物群落的动态，特别是产甲烷菌群落。在本研究中，从 7 对设计并在不同浓度和条件下进行测试的引物中选择了 3 对引物，通过基于 SYBR Green 系统的实时 PCR 检测 Methanosarcina、Methanoculleus 和 Methanobacterium。通过标准曲线的高度线性关系证明了所开发方法的功能，通过测试产甲烷和非产甲烷菌株分离的 DNA 经验证了每个引物的特异性。这些测定还表现出良好的重复性和再现性，这表明该方法具有稳健性。所描述的引物成功地用于研究来自厌氧共消化的 10 个样品中的产甲烷菌群。产甲烷菌属 Methanosarcina 是主要的产甲烷菌群。
