KEGG pathway enrichment analysis of DEGs (differentially expressed genes) indicated primordium initiation was significantly related to 66 pathways, such as “Ribosome”, “metabolism of xenobiotics by cytochrome P450”, and “glutathione metabolism” (among others). KEGG and KOG analyses respectively mapped 32,765 unigenes to 202 pathways and 19,408 unigenes to 25 categories. The de novo assembly generated a total of 48,549 unigenes, of which 71.53% (34,728) unigenes could be annotated by at least one of the KEGG (Kyoto Encyclopedia of Genes and Genomes), GO (Gene Ontology), and KOG (Eukaryotic Orthologous Group) databases. To investigate this process, 4.8 × 10⁸ RNA-Seq reads were acquired from three stages: hyphal knot (SM), primordium (SP), and primordium differentiation (SPD). The genes associated with fruiting body formation of Sparasis latifolia are valuable for improving mushroom breeding. Our study provides important insights into the molecular mechanisms underlying fruiting body development and sporulation in G. Meanwhile, highly expressed unigenes related to meiosis such as DMC1, MSH4, HOP1, and Mek1 were also analyzed. In addition, highly expressed unigenes associated with the transition between different developmental stages were identified, including those encoding hydrophobins, cytochrome P450s, extracellular proteases, and several transcription factors. In contrast, DEGs identified in G2 versus G3 were mainly assigned to the category of metabolism of amino acids and their derivatives, comprising mostly upregulated unigenes. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of DEGs identified in G1 versus G2 revealed that, in addition to global and overview maps, enriched pathways were related to amino acid metabolism and carbohydrate metabolism. Differentially expressed genes (DEGs) were identified between the evaluated stages: 3135 DEGs in G1 versus G2 120 in G2 versus G3 3919 in G3 versus G in G4 versus G5. Functional annotation of transcriptomes matched 72.49% of the unigenes to known proteins available in at least one database.

In total, 26,137 unigenes with an average length of 1078 bp were de novo assembled. lucidum, transcriptome analysis based on RNA sequencing was carried out on different developmental stages: mycelium (G1) primordium (G2) young fruiting body (G3) mature fruiting body (G4) fruiting body in post-sporulation stage (G5). To better understand the molecular mechanisms underlying fruiting body development in G. Ganoderma lucidum is an edible mushroom highly regarded in the traditional Chinese medicine.

Primordia hyphal knots