This study was designed to assess the influence of three soil DNA extraction procedures, namely the International Organization for Standardization (ISO-11063, GnS-GII and modified ISO procedure (ISOm), on the taxonomic diversity and composition of soil bacterial and fungal communities. The efficacy of each soil DNA extraction method was assessed on five soils, differing in their physico-chemical characteristics and land use. A meta-barcoded pyrosequencing approach targeting 16S and 18S rRNA genes was applied to characterize soil microbial communities. We first observed that the GnS-GII introduced some heterogeneity in bacterial composition between replicates. Then, although no major difference was observed between extraction procedures for soil bacterial diversity, we saw that the number of fungal genera could be underestimated by the ISO-11063. In particular, this procedure underestimated the detection in several soils of the genera Cryptococcus, Pseudallescheria, Hypocrea and Plectosphaerella, which are of ecological interest. Based on these results, we recommend using the ISOm method for studies focusing on both the bacterial and fungal communities. Indeed, the ISOm procedure provides a better evaluation of bacterial and fungal communities and is limited to the modification of the mechanical lysis step of the existing ISO-11063 standard.

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Wastewater can be recycled in agricultural soil as fertilizer to increase crop yields. However, adding wastewater induces sometimes ecotoxicological issues such as pollution by toxic compounds, which may lead to the loss of arable land. Bioenergy crops such as Miscanthus × giganteus have been tested to rehabilitate polluted soils, but the impact of Miscanthus on soil microbes is unknown. Here, we evaluated the effects of Miscanthus cropping on bacterial and fungal taxonomic composition in a wastewater-contaminated soil using synchronic and diachronic evaluation strategies. A 3-year field experiment close to Paris was set up on an agricultural site irrigated by raw wastewater for more than one century, thus resulting in strong metal and organic contamination. Soil microbial taxonomic composition was characterized by direct analysis of soil DNA using metagenomic tools such as 454 pyrosequencing of ribosomal genes. Our results demonstrate that Miscanthus cropping stimulates specific populations of bacteria such as Rhizobiales, increased by 1.4 in relative abundance, Nistrospira (x1.5), Azospira (x2), and Gemmatimonas (x2), and fungi: Glomeromycota (x3) and Mortierella (x1.5) for fungi. Noteworthy, these microbial genera are known to be strongly involved in plant symbiosis, organic matter mineralization, and nutrient cycling. Overall our findings show that Miscanthus cropping enhances regeneration of soil microbiological functions and services in polluted soil by stimulating populations beneficial for soil fertility and crop production