Genomics DNA from the samples were extracted via MP Fast DNA SPIN Kit for Soil Kit (MP Biomedical). Qubit 3.0 fluorometer was used to determine the concentration of DNA, and a 0.8% agar gel electrophoresis buffer was used to detect DNA integrity [10 (link)].
The V3 and V4 regions of bacterial 16S rRNA and fungal ITS rRNA were amplified based on previous research in our laboratory [11 (link)].
The PCR amplification system and conditions have been modified according to previous methods [12 (link)]. PCR reactions were performed in triplicate using 0.4 μL of FastPfu polymerase, 4 μL of 5 × FastPfu buffer, 0.8 μL (5 μM) of each primer, 2 μL of 2.5 mM dNTPs, and 10 ng of template DNA under the following conditions: 95 °C for 3 min, 27 cycles at 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 45 s, and a final extension at 72 °C for 5 min.
The V3 and V4 regions of bacterial 16S rRNA and fungal ITS rRNA were amplified based on previous research in our laboratory [11 (link)].
The PCR amplification system and conditions have been modified according to previous methods [12 (link)]. PCR reactions were performed in triplicate using 0.4 μL of FastPfu polymerase, 4 μL of 5 × FastPfu buffer, 0.8 μL (5 μM) of each primer, 2 μL of 2.5 mM dNTPs, and 10 ng of template DNA under the following conditions: 95 °C for 3 min, 27 cycles at 95 °C for 30 s, 55 °C for 30 s, and 72 °C for 45 s, and a final extension at 72 °C for 5 min.