Sample from the 08th year of restoration (repeat 2)
Genomic DNA was extracted from 0.5 g of homogenized soil using the DNeasy PowerSoil Kit (Qiagen) following the manufacturers protocol. The V3V4 region of the 16S rRNA gene of rhizosphere bacteria was amplified using the 16sF/16sR primers. PCR conditions included initial denaturation at 95C for 3 min, followed by 25 cycles of denaturation at 95C for 30 sec, annealing at 55C for 30 sec, and extension at 72C for 30 sec, with a final elongation at 72C for 5 min. The amplified PCR products were purified and indexed using the Illumina Nextera XT Library Prep Kit, followed by sequencing on the Illumina MiSeq platform with a 2  307 bp paired-end read configuration.