A rapid microtiter plate method to measure carbon dioxide evolved from carbon substrate amendments so as to determine the physiological profiles of soil microbial communities by using whole soil
A rapid microtiter plate method to measure carbon dioxide evolved from carbon substrate amendments so as to determine the physiological profiles of soil microbial communities by using whole soil
复制标题
DOI：
10.1128/aem.69.6.3593-3599.2003
复制
发表时间：
2003-06-01
影响因子：
4.4
通讯作者：
Potts, JM
中科院分区：
文献类型：
--
作者：
Campbell, CD;Chapman, SJ;Potts, JM
单一碳源测试（Biolog），旨在识别细菌，已成为非常流行的代谢指纹土壤微生物群落，尽管与使用碳源配置文件，主要选择快速生长的细菌相关的缺点。在本文中，我们描述了一种替代方法的使用，该方法结合了Biolog社区水平生理剖面（CLPP）方法的优点，其中使用基于微量滴定的检测板，能够测量整个土壤中二氧化碳的演变。该方法便于在短时间内（4至6小时）进行测量，并且不需要提取和培养生物体。使用深孔微量滴定板作为测试威尔斯孔，将土壤置于其中。描述了填充深孔板并将其与第二可移除检测板接合的设备。介绍了两种检测系统，一种是在碱吸收剂中的简单比色反应，另一种是用放射性碳源进行闪烁计数。通过使用不同植被类型下的土壤和用废水污泥处理的土壤，将所述方法与Biolog-CLPP系统进行比较。我们的目的是测试的假设，使用整个土壤将有特定的优势，使用提取物，可以得到更直接的反应，反映活动，而不是增长的基板。全土法更快速，能更早地检测出碳源的使用。此外，所获得的代谢指纹可以区分污泥处理。
Sole-carbon-source tests (Biolog), designed to identify bacteria, have become very popular for metabolically fingerprinting soil microbial communities, despite disadvantages associated with the use of carbon source profiles that primarily select for fast-growing bacteria. In this paper we describe the use of an alternative method that combines the advantages of the Biolog community-level physiological profile (CLPP) method, in which microtiter-based detection plates are used, with the ability to measure carbon dioxide evolution from whole soil. This method facilitates measurement over short periods of time (4 to 6 h) and does not require the extraction and culturing of organisms. Deep-well microtiter plates are used as test wells into which soil is placed. The apparatus to fill the deep-well plates and interface it with a second removable detection plate is described. Two detection systems, a simple colorimetric reaction in absorbent alkali and scintillation counting with radioactive carbon sources, are described. The methods were compared to the Biolog-CLPP system by using soils under different vegetation types and soil treated with wastewater sludge. We aimed to test the hypothesis that using whole soil would have specific advantages over using extracts in that more immediate responses to substrates could be obtained that would reflect activity rather than growth. The whole-soil method was more rapid and gave earlier detection of C source use. Also, the metabolic fingerprints obtained could discriminate between sludge treatments.