Illumina amplicon sequencing of 16S rRNA genes (bacteria): rhizosphere
Total DNAwas extracted from bulk soil (0.25 g) and the rhizosphere and root fractions (0.25 g when possible and the entire material available when quantity was less than 0.25 g). DNA extraction was performed using PowerSoil-htp96Well Soil DNA Isolation Kit according to manufacturers instructions (MOBIO Laboratories, UK). Bacterial community biodiversity was assessed using Illumina amplicon sequencing of 16S rRNA genes (bacteria) to phylogenetically identify responsive taxa. Amplicon libraries were constructed according to the dual indexing strategy of Kozich et al. (2013), with each primer consisting of the appropriate Illumina adapter, an 8-nt index sequence, a 10-nt pad sequence, a 2-nt linker and the gene specific primer. For 16S, the V3-V4 hypervariable regions of the 16S rRNA gene was targeted using primers based upon the universal primer sequence 341F and 806R. (see Merino-Martn et al. 2020. Rhizosphere bacteria are more strongly related to plant root traits than fungi in temperate montane forests: insights from closed and open forest patches along an elevational gradient. Plant and Soil 450, 183200. https://doi.org/10.1007/s11104-020-04479-3. and Merino-Martn et al. 2020. Interacting effects of land use type, microbes and plant traits on soil aggregate stability. Soil Biology and Biochemistry 154 (2021) 108072).
SUB9976613
=_xlfn.CONCAT("FIXSOIL_16S_",A12)