An integrated analytical approach for assessing the biological status of the soil microbial community

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• 作者：Alice Pascaud ; Marie-Louise Soulas ; Samira Amellal ; Guy Soulas ; ^{guy.soulas@oenologie.u-bordeaux2.fr} ; ^{guy.soulas0001@orange.fr}
• 关键词：Density-gradient centrifugation ; ATP ; Dehydrogenase activity ; DHA ; Physiological profiles ; Species diversity ; Soil biological status
• 刊名：European Journal of Soil Biology
• 出版年：2012
• 期刊代码：43_11645563
• 类别：abs
• 出版时间：March-April, 2012
• 卷：49
• 期：Complete
• 页码：98-106
• 文件大小：385 K

摘要

An integrated multicriteria analytical procedure for rapid, cost-effective characterisation of the biological status of soil bacterial community was developped. Commercially-available, light emission-based bioassays were selected for measuring cell density, activity, and diversity. All but Terminal Restriction Fragment Length Polymorphism (T-RFLP) were designed for multiwell-plate formats and high-throughput screening potential. Adenosine Tri Phosphate (ATP) was measured using a bioluminescence assay. Dehydrogenase activity (DHA) was measured on growing cells. Kinetic measurements of the formation of a coloured formazan derivative was used after nutrient broth addition to estimate initial cell concentrations by reference to Escherichia coli added as internal standard. Compared to conventional ATP and DHA determinations in soils, the procedures described here do not require extraction of ATP or formazan derivative from the soil matrix. Metabolic diversity was characterised using the Biolog鈩?system. T-RFLP was chosen for assessing bacterial community structure. The bioassays were performed on microbial preparations obtained after either direct dilution of soil suspensions or prior density-gradient separation of microbial cells from the soil matrix. Dilution maintains the original structure of native dominant microbial communities. Density-gradient separation of microbial cells is highly selective, drastically modifying metabolic (CLP Profiles) and species (T-RFLP patterns) diversity, as well as activity parameters.