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. The activity of microorganisms in soil is important for a robustfunctioning of soil and related ecosystem services. Hence, there is anecessity to identify the composition, diversity, and function of the soilmicrobiome in order to determine its natural properties, functioning, andoperating range as well as to assess ecotoxicological effects due toanthropogenic activities. Numerous microbiological methods currently existin the literature and new, more advanced methods continue to be developed;however, only a limited number of these methods are standardised.Consequently, there is a need to identify the most promisingnon-standardised methods for assessing soil quality and to transform theminto standards. In agreement with the “Ecosystem Service Approach”, newmethods should focus more on soil microbial functions, including nutrientcycling and greenhouse gas emission, pest control and plant growthpromotion, carbon cycling and sequestration, as well as soil structuredevelopment and filter function. The few existing standardised methodsavailable that focus on the function of the soil microbiome mostly includemeasurements, like basal respiration, enzyme activities, and biodegradationof organic matter, under well-defined conditions in the lab. This paper setsout to summarise and expand on recent discussions within the InternationalOrganization for Standardization (ISO), Soil Quality – BiologicalCharacterization sub-committee (ISO TC 190/SC 4), where a need was identifiedto develop scientifically sound methods which would best fulfil thepractical needs of future users for assessing soil quality, going beyond theexisting test systems. Of particular note is the current evolution ofmolecular methods in microbial ecology that use quantitative real-time PCR(qPCR) to produce a large number of new functional endpoints which are moresensitive as compared to “classical” methods. Quantitative PCR assesses theabundance of microbes that catalyse major transformation steps in nitrogenand phosphorus cycling, greenhouse gas emissions, chemical transformationsincluding pesticide degradation, and plant growth promotion pathways basedon the assessment of marker gene sequences that drive the related processes.In the assessment of soil quality methods, it was found that most methodsfocus on bacteria and related endpoints. Techniques to describe fungalcommunities as well as their functional traits are far less represented. Assuch, techniques to analyse fungal enzyme activities are proposed.Additionally, methods for the determination of microbial growth rates andefficiencies, including the use of glomalin as a biochemical marker for soilaggregation, are discussed. Furthermore, field methods indicative of carbonturnover, including the litter bag test and a modification to the tea bagtest, are presented. However, it is obvious that with increasingdevelopments in high throughput sequencing technologies and big dataanalyses, including metagenomics analysis, it will be possible to implementthese technologies into the standardisation process for assessing thefunctions of the soil microbiome. Overall, it is suggested that endpointsshould represent a potential function of soil microorganisms rather thanactual activity levels, as the latter can largely be dependent on short-termvariable soil properties such as pedoclimatic conditions, nutrientavailability, and anthropogenic soil cultivation activities.

Identification of new microbial functional standards for soil quality assessment