Illumina 16S metabarcoding of DNA-3BS-LB10%
LB 10% NaCl was prepared in the conventional method but with modifications (10 g of tryptone, 100 g of NaCl, 5 g of yeast extract, 15 g of agar and 1 liter of distilled water), sterilized by autoclaving and subsequently plated in petri dishes. To obtain the culturable microbiota from the BS and RZ samples, a suspension of 1 g of soil in 1 mL of 1X Phosphate-Buffered Saline (PBS 1X) was used as inoculum. The mixture was homogenized by shaking in the Revolver Rotator (LabNet) for 2 hours and then decanted by gravity for 30 minutes at room temperature. 100 uL of supernatant were inoculated into LB 10% NaCl plates and distributed in the petri dish with an L-shaped cell Spreader. Subsequently, the plates were incubated at 30C for 72 h, and after this time, the bacterial growth was resuspended in 200 uL of 1X PBS to perform DNA isolation using the DNeasy Blood & Tissue kit (Qiagen), following the manufacturer's recommendations. The V1-V3 region of the bacterial 16S rRNA gene was amplified with the primers 28F (5-GAGTTTGATCNTGGCTCAG-3) and 519R (5-GTNTTACNGCGGCKGCTG-3). The amplicons were sequenced with an Illumina sequencer using the service of Molecular Research LP. (Mr. DNA Lab). In the case of the RNA samples, reverse transcription was performed prior to amplification to generate cDNA.