Rhizosphere metagenome of Oryza sativa:16S metabarcoding Bama site
Less than 24h after sampling, root samples (rice roots including rhizosphere) stored at 4C were processed. In order to separate the different root compartments, the tubes were vortexed vigorously one minute and then, roots were removed from the PBS solution using sterile forceps. The remaining PBS solution was considered as the rhizosphere compartment. DNA extraction from the rhizosphere were performed on the same day as the process of compartment separation. 0.25g of rhizosphere was extracted using the DNeasy PowerSoil Kit (Qiagen, Hilden, Germany), following manufacturers recommendations. DNA quality and quantity was verified using a NanoDrop ND-1000 spectrophotometer. PCR amplification, library and MiSeq Illumina sequencing were performed by Macrogen (Seoul, South Korea) using primers 341F (16S V3F, 5-CCTACGGGNGGCWGCAG-3) and 785R (16S V4R, 5-GACTACHVGGGTATCTAATCC-3) to amplify the V3 -V4 regions of the 16S rRNA gene.