design: Total RNA was extracted from axenic cultures using the TRIzol Reagent / RNeasy Mini Kit (Qiagen). RNA integrity and quantity were assessed using an Agilent 2100/2200 Bioanalyzer, NanoDrop, and 1% agarose gel electrophoresis. For library construction, 1 g of total RNA per sample was subjected to ribosomal RNA (rRNA) depletion using a dedicated rRNA removal kit. The rRNA-depleted RNA was then fragmented and reverse-transcribed into first-strand cDNA using ProtoScript II Reverse Transcriptase with random primers in the presence of Actinomycin D. Second-strand cDNA was synthesized using a Second Strand Synthesis Enzyme Mix containing dUTP instead of dTTP to achieve strand specificity. The double-stranded cDNA was purified, end-repaired, A-tailed, and ligated to Illumina adapters. Adapter-ligated fragments of ~400 bp (insert size ~300 bp) were size-selected. The dUTP-marked second strand was digested with Uracil-Specific Excision Reagent (USER) enzyme to ensure strand orientation. Finally, the libraries were amplified with index-containing P5 and P7 primers, purified, and validated for size distribution and concentration using Qsep100 and Qubit 3.2 Fluorometer, respectively.