Metatranscriptome of soil: Untreated soil (Control)-T1a
Total RNA extracted from the soil samples was subjected to ribosomal RNA (rRNA) depletion using the NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7860L) according to the manufacturer's instructions. The rRNA-depleted RNA was subsequently used for library preparation with the NEBNext UltraTM II Directional RNA Library Prep Kit for Illumina (NEB #E7765L) following the manufacturer's protocol. Briefly, the rRNA-depleted RNA was fragmented, followed by first-strand and second-strand cDNA synthesis. The resulting double- stranded cDNA underwent end repair, adapter ligation, and PCR amplification to generate the final sequencing libraries. The prepared libraries were quantified using the Qubit 4 Fluorometer (Invitrogen, Thermo Fisher Scientific, USA) and the QuantStudio Real-Time PCR System (Thermo Fisher Scientific, USA). Library fragment size distribution was assessed using the Agilent 2100 Fragment Analyzer System (Agilent Technologies, USA). Libraries with an insert size of approximately 300350 bp and an effective concentration greater than 10 nM were pooled and sequenced on the Illumina NovaSeq 6000 platform (Illumina Inc., USA) using a 150 bp paired-end sequencing strategy (2  150 bp).