Introduction: The genus Laccaria has been reported from temperate and tropical areas and is an important constituent in forest ecosystems. However, the species diversity of Laccaria in Southwest China (Yunnan) has been underestimated.Methods: In this paper, descriptions based on morphological and multi-gene sequence data from internal transcribed spacer (ITS) region, large subunit ribosomal RNA gene (nrLSU), translation elongation factor 1-α (TEF1α) and the polymerase II second largest subunit (RPB2) of three new Laccaria species from Southwest China (Yunnan) are reported.Results: Two of these were characterized by orange pileus and globose to subglobose basidiospores: L. cinnabarina and L. spinulosa. While L. cinnabarina has orange red colored basidiocarps with conspicuously pellucid-striate pattern, and a fibrillose stipe with longitudinally striations, L. spinulosa has a brownish orange to brown fruiting body with light white pruinae and 2-spored basidia. Laccaria longistriata is characterized by brown to flesh-colored basidioma, prominently striate to sulcate pileus and globose to subglobose basidiospores.Discussion: The three new species were described, illustrated and compared with closely related species in morphology and phylogeny.

Long-term in vitro culture of human mesenchymal stem cells (MSCs) leads to cell lifespan shortening and growth stagnation due to cell senescence. Here, using sequencing data generated in the public domain, we have established a specific regulatory network of “transcription factor (TF)-microRNA (miRNA)-Target” to provide key molecules for evaluating the passage-dependent replicative senescence of mesenchymal stem cells for the quality control and status evaluation of mesenchymal stem cells prepared by different procedures. Short time-series expression miner (STEM) analysis was performed on the RNA-seq and miRNA-seq databases of mesenchymal stem cells from various passages to reveal the dynamic passage-related changes of miRNAs and mRNAs. Potential miRNA targets were predicted using seven miRNA target prediction databases, including TargetScan, miRTarBase, miRDB, miRWalk, RNA22, RNAinter, and TargetMiner. Then use the TransmiR v2.0 database to obtain experimental-supported transcription factor for regulating the selected miRNA. More than ten sequencing data related to mesenchymal stem cells or mesenchymal stem cells reprogramming were used to validate key miRNAs and mRNAs. And gene set variation analysis (GSVA) was performed to calculate the passage-dependent signature. The results showed that during the passage of mesenchymal stem cells, a total of 29 miRNAs were gradually downregulated and 210 mRNA were gradually upregulated. Enrichment analysis showed that the 29 miRNAs acted as multipotent regulatory factors of stem cells and participated in a variety of signaling pathways, including TGF-beta, HIPPO and oxygen related pathways. 210 mRNAs were involved in cell senescence. According to the target prediction results, the targets of these key miRNAs and mRNAs intersect to form a regulatory network of “TF-miRNA-Target” related to replicative senescence of cultured mesenchymal stem cells, across 35 transcription factor, 7 miRNAs (has-mir-454-3p, has-mir-196b-5p, has-mir-130b-5p, has-mir-1271-5p, has-let-7i-5p, has-let-7a-5p, and has-let-7b-5p) and 7 predicted targets (PRUNE2, DIO2, CPA4, PRKAA2, DMD, DDAH1, and GATA6). This network was further validated by analyzing datasets from a variety of mesenchymal stem cells subculture and lineage reprogramming studies, as well as qPCR analysis of early passages mesenchymal stem cells versus mesenchymal stem cells with senescence morphologies (SA-β-Gal⁺). The “TF-miRNA-Target” regulatory network constructed in this study reveals the functional mechanism of miRNAs in promoting the senescence of MSCs during in vitro expansion and provides indicators for monitoring the quality of functional mesenchymal stem cells during the preparation and clinical application.

Species of Entoloma are exceptionally diverse and widely distributed, with cuboid-spored members occurring mainly in subtropical to tropical regions. These cuboid-spored taxa are primarily classified into two subgenera, E. subg. Cuboeccilia and E. subg. Cubospora. During surveys conducted in Yunnan Province, a representative subtropical region of China, four new species belonging to E. subg. Cubospora were identified through multigene phylogenetic analyses (ITS, LSU, rpb2 , and tef1 ) combined with detailed morphological observations. Entoloma acutiflavum is distinguished by its yellow basidiomata, papillate pileus, heterogeneous lamellar edge, and the presence of clamp connections. Entoloma bichromum can be recognized by a centrally papillate pileus, serrate pileal and lamellar margins, and clavate to cylindrical pleurocystidia that are flexuous, furcate, or occasionally ventricose. Entoloma guttuliferum is characterized by its yellowish-white to white papillate pileus, clavate to elongated clavate terminal cells in both the pileipellis and stipitipellis, and pigmented cystidia, lamellar trama, pileipellis, and stipitipellis. Entoloma rufosquamulosum is identified by its reddish-brown to grayish-violet squamose pileus, with reddish-brown pigments present in the upper hyphal layer and terminal cells of the pileipellis. Detailed descriptions, line drawings, habitat photographs, SEM micrographs, and comparisons with morphologically and phylogenetically related taxa are provided. A key to the species of Entoloma subg. Cubospora from China is also included.