Grafting and Microbial Consortia Modulate Rhizosphere Microbiome and Tomato Productivity under Mediterranean Greenhouse
DNA extraction was carried out following the manufacturers practice provided by Soil DNA Isolation Plus Kit (Norgen Biotek, Thorol, Canada). Amplicon Generation 16S and ITS genes of distinct regions (16SV4/16SV3/16SV3 and ITS1) were amplified used specific primer with the barcode. All PCR reactions were carried out with 15 L of Phusion High- Fidelity PCR Master Mix; 0.2 M of forward and reverse primers, and about 10 ng template DNA. Thermal cycling consisted of initial denaturation at 98 for 1 min, followed by 30 cycles of denaturation at 98 for 10 s, annealing at 50 for 30 s, and elongation at 72for 30 s and 72for 5 min.The PCR products were purified using magnetic bead purification. Samples were mixed in equidensity ratios based on the concentration of PCR products. After thorough mixing, the PCR products were detected and target bands were recovered Sequencing libraries were generated and indexes were added. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina platforms, according to effective library concentration and data amount required