Soil DNA Extraction and Microbial Sequencing

The soil samples were collected to extract DNA according to the FastDNA^TM SPIN Kit (MP Biomedicals, USA)for Soil extraction specification, and agarose gel electrophoresis was used to detect DNA quality. DNA concentration detector One drop 2000 (Thermo, USA) was used to determine the concentration of the DNA. The bacterial 16S rRNA sequencing fragment was V3-V4, and the regional universal primer was 338 F:5’-ACTCCTACGGGAGGCAGCAG-3’, 806 R: 5’-GGACTACHVGGGTWTCTAAT-3’,^{16 (link)} the fungal ITS sequencing fragment was ITS2, and the regional universal primer was fITS7: 5’-GTGARTCATCGAATCTTTG-3’, ITS4: 5’-TCCTCCGCTTATTGATATGC-3’,^{17 (link)} PCR reaction system and program were performed formed following a previous study.^{18 (link)} PCR products were purifed with Gel Extraction Kit (Vazyme, China) and pooled in equimolar concentrations. Paired-end sequencing (PE300) of bacterial and fungal amplicons were carried out on an Illumina MiSeq platform at Hangzhou Lianchuan Biotechnology Co., Ltd (Hangzhou, China).