Transcription factors (TFs) play a crucial role in gene expression, and studying them can lay the foundation for future research on the functional characterization of TFs involved in various biological processes. In this study, we conducted a genome-wide identification and analysis of TFs in the thermotolerant basidiomycete fungus, Coriolopsis trogii. The TF repertoire of C. trogii consisted of 350 TFs, with C2H2 and Zn2C6 being the largest TF families. When the mycelia of C. trogii were cultured on PDA and transferred from 25 to 35 °C, 14 TFs were up-regulated and 14 TFs were down-regulated. By analyzing RNA-seq data from mycelia cultured at different temperatures and under different carbon sources, we identified 22 TFs that were differentially expressed in more than three comparisons. Co-expression analysis revealed that seven differentially expressed TFs, including four Zn2C6s, one Hap4_Hap_bind, one HMG_box, and one Zinc_knuckle, showed significant correlation with 729 targeted genes. Overall, this study provides a comprehensive characterization of the TF family and systematically screens TFs involved in the high-temperature adaptation of C. trogii, laying the groundwork for further research into the specific roles of TFs in the heat tolerance mechanisms of filamentous fungi.

BackgroundIt is generally accepted that nuclear genes in eukaryotes are located independently on chromosomes and expressed in a monocistronic manner. However, accumulating evidence suggests a more complex landscape of gene structure and transcription. Ganoderma lingzhi, a model medicinal fungus, currently lacks high-quality genome annotation, hindering genetic studies.ResultsHere, we reported a golden annotation of G. lingzhi, featuring 14,147 high-confidence genes derived from extensive manual corrections. Novel characteristics of gene structure and transcription were identified accordingly. Notably, non-canonical splicing sites accounted for 1.99% of the whole genome, with the predominant types being GC-AG (1.85%), GT-AC (0.05%), and GT-GG (0.04%). 1165 pairs of genes were found to have overlapped transcribed regions, and 92.19% of which showed opposite directions of gene transcription. A total of 5,412,158 genetic variations were identified among 13 G. lingzhi strains, and the manually corrected gene sets resulted in enhanced functional annotation of these variations. More than 60% of G. lingzhi genes were alternatively spliced. In addition, we found that two or more protein-coding genes (PCGs) can be transcribed into a single RNA molecule, referred to as polycistronic genes. In total, 1272 polycistronic genes associated with 2815 PCGs were identified.ConclusionsThe widespread presence of polycistronic genes in G. lingzhi strongly complements the theory that polycistron is also present in eukaryotic genomes. The extraordinary gene structure and transcriptional activity uncovered through this golden annotation provide implications for the study of genes, genomes, and related studies in G. lingzhi and other eukaryotes.

Ganoderma lucidum has a wide carbon spectrum, while the expression profile of key genes relevant to carbon metabolism on different carbon sources has been seldom studied. Here, the transcriptomes of G. lucidum mycelia cultured on each of 19 carbon sources were conducted. In comparison with glucose, 16 to 1,006 genes were upregulated and 7 to 1,865 genes were downregulated. Significant gene expression dynamics and induced activity were observed in laccase genes when using agricultural and forestry residues (AFRs) as solo carbon sources. Furthermore, study of laccase gene family in two haploids of G. lucidum GL0102 was conducted. Totally, 15 and 16 laccase genes were identified in GL0102_53 and GL0102_8, respectively, among which 15 pairs were allelic genes. Gene structures were conserved between allelic laccase genes, while sequence variations (most were SNPs) existed. Nine laccase genes rarely expressed on all the tested carbon sources, while the other seven genes showed high expression level on AFRs, especially Gllac2 and Gllac7 , which showed 5- to 1,149-fold and 4- to 94-fold upregulation in mycelia cultured for 5 days, respectively. The expression of H53lac7 was consistently higher than that of H8lac7_1 on all the carbon sources except XM, exhibiting a case of allelic expression bias. A total of 47 SNPs and 3 insertions/deletions were observed between promoters of H53lac7 and H8lac7_1 , which lead to differences in predicted binding sites of zinc fingers. These results provide scientific data for understanding the gene expression profile and regulatory role on different carbon sources and may support further functional research of laccase.

Amomum maximum Roxb. rhizome is a fork medicine mainly used in South and Southeast Asia. In present study, the hypoglycaemic effects of the ethanolic extract of A. maximum rhizome were demonstrated both on α-glucosidase assay in vitro and streptozotocin (STZ)-induced postprandial hyperglycaemia in mice. Furthermore, six labdane diterpenes, amoxanthin A (1), ottensinin (2), coronarin D (3), coronarin D methyl ether (4), isocoronarin D (5), and zerumin (6), were isolated from its ethyl acetate sub-fraction with the guidance of α-glucosidase inhibitory activity. Among these compounds, 2 and 6 exhibited significant inhibitory effect on α-glucosidase, as well as on STZ-induced high postprandial blood glucose levels in mice. Additionally, molecular docking analysis revealed that 2 and 6 could firmly bind to the active sites of α-glucosidase. These results suggest that compounds 2 and 6 are the main anti-hyperglycaemic agents present in A. maximum, which may demonstrate potential beneficial effects in diabetes management.

硬毛粗盖孔菌是一种结实能力强的耐热真菌,为了探究其过氧化氢酶(CAT)基因家族的基本特征和功能,分别对不同交配型单核体Ct001 29和Ct001 31基因组的CAT基因家族进行了鉴定,并分析了不同CAT家族基因在不同温度条件下培养的菌丝、原基和子实体中的表达特征.硬毛粗盖孔菌基因组上有3个CAT基因(Ctcat1-Ctcat3),编码510-743个氨基酸;Ctcat1、Ctcat2和Ctcat3的等位基因之间的结构及序列相对保守,但编码区也存在少量SNP变异(6-14个).在25℃培养菌丝和35℃培养菌丝、原基及子实体中,35℃菌丝的CAT酶活最高,为278 U/mg蛋白,原基期CAT酶活最低,为4 U/mg蛋白.Ctcat2的表达丰度显著高于Ctcat1和Ctcat3,Ctcat1和Ctcat3具有相似的表达模式,在35℃C菌丝、原基和子实体中均上调表达;而Ctcat2具有相反的表达模式,在35℃菌丝、原基和子实体中均下调表达.此外,Ctcat2在原基时期、Ctcat3在25℃菌丝和35℃菌丝中具有偏好表达,Ct29cat2和Ct29cat3的表达量均高于各自的等位基因.本研究所发现的CAT等位基因表达偏好为进一步揭示大型真菌CAT的基因功能奠定了基础.

A novel sugarcane juice beverage was successfully produced by liquid fermentation of G. lucidum and homogenate process with selecting the day 10 fermentation broth as the raw materials. The beverage was evaluated for its sensory, nutritional and antioxidant activities. The results showed that the beverage by the G.lucidum fermentation was rich in polysaccharides, triterpenes, proteins, and flavonoids, which were all significantly higher than sugarcane juice, especially polysaccharides and triterpenoids. And the beverage could retain the sensory quality of sugarcane juice and could therefore be favored by the consumers. The in vitro antioxidant activity showed that the beverage had high scavenging and reducing abilities. DPPH and ABTS•+ radical scavenging rate of 1 mL fermented beverage were similar to 0.04 mg/mL VC, while the hydroxyl radical scavenging rate and reducing power of 1 mL of fermented beverage were similar to 1.68 mg/mL and 0.19 mg/mL VC, respectively. Taken together, the sugarcane juice fermented beverage was a natural flavor substances with high antioxidant activity and non-toxicity.

对花脸香蘑(Lepista sordida)的过氧化氢酶(catalase,CAT)基因进行克隆鉴定和生物信息学分析,并分别对花脸香蘑菌株LS01和LS02菌丝在不同培养时间(5、10、15、20d)和35℃高温处理3、48 h的胞内H2O2含量、CAT活性和CAT基因表达量进行测定.结果表明:4个花脸香蘑CAT基因(Lscat1～Lscat4)编码527～729个氨基酸;两个菌株的Lscat1～Lscat4分别编码相同数量的氨基酸,具有一定的遗传差异,也具有相似的基因结构;随着培养时间增加,两个菌株菌丝胞内H2O2含量呈逐渐上升趋势,而不同培养时间CAT活性无显著差异.两个菌株菌丝中的Lscat1～Lscat4在不同培养时间表达量具有差异.与对照比较,35℃处理3 h时菌株LS01和LS02菌丝胞内H2O2含量无显著变化;35℃处理48 h时菌株LS01胞内H2O2含量显著升高,菌株LS02胞内H2O2含量显著下降;35℃处理3 h时两个菌株CAT活性均升高,35℃处理48 h时两个菌株CAT活性均降低.与对照比较,35℃处理3 h时两个菌株的Lscat1、Lscat2和Lscat3均下调表达;35℃处理48 h时,Lscat2和Lscat3在菌株LS01中下调表达,Lscat1、Lscat2和Lscat3在LS02中均上调表达;35℃处理3 h时,Lscat4在菌株LS01中无显著变化,在菌株LS02中下调表达;35℃处理48 h时,Lscat4在两个菌株中均极显著上调表达.研究结果为进一步发掘花脸香蘑CAT基因功能提供参考.

Although triterpenoids are one of the main active ingredients in Ganoderma lucidum, their accumulation and antioxidant activity during the different developmental stages of G. lucidum cultivation in bagasse remains unclear. In this study, we investigated the content and antioxidant activity of total triterpenoids extracted from G. lucidum strain GL102 during the five growth stages. The obtained results showed that the highest content (12.06 mg/g) was detected in stage 3 (young fruiting body), similar to the contents of ganoderic acids B and G. However, ganoderic acids A and D exhibited maximal contents in stage 5 (spore-ejected fruiting body). The triterpenoids extracted during stage 5 were most capable of scavenging DPPH, OH, and ABTS(+) radicals, with scavenging rates of 65.88%, 86.45%, and 97.91%, respectively. Based on in vivo antioxidant assays conducted on zebrafish, the safe concentration of these triterpenoids was 0.03 mg/mL. At this concentration, the G. lucidum triterpenoids extracted during stage 5 could decrease lipopolysaccharide-induced intracellular reactive oxygen species to a level that was nearly normal (similar to the control group). The accumulation profile and antioxidant activity results reported herein provide the scientific basis needed to promote the utilization of triterpenoids derived from bagasse-cultivated G. lucidum.

对适用于花脸香蘑(Lepista sordida)荧光定量PCR内参基因进行筛选,并检测花脸香蘑高产胞外多糖诱变菌株LS01_ 10_9及其出发菌株LS01_10多糖合成途径关键基因和多糖代谢途径基因表达量.结果 表明:核糖体蛋白L4(ribosomal protein L4,rpl4)基因和3-磷酸甘油脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,gapdh)基因表达丰度高、扩增效率较好且稳定,是花脸香蘑荧光定量PCR理想的内参基因.与出发菌株LS01_10比较,高产胞外多糖诱变菌株LS01_ 10_9中5个参与多糖合成途径的关键基因GK、GMD-3、UGE、UGDG、PFK表达量显著增加,4个参与多糖代谢途径的基因GT-1、GT-2、NOX-3、NOX-4表达量显著增加.

为了筛选出最适合灵芝菌种复壮的培养基和灵芝菌种的保藏方法,通过添加不同含量的甘蔗渣提取液作为灵芝菌株复壮的培养基,观察灵芝菌丝在不同培养基下的生长速度和生长状况,并通过对灵芝菌种酶活力进行测定分析,比较几种不同保藏方法对灵芝菌种活力的影响.结果表明:马铃薯葡萄糖培养基(PDA)培养基添加甘蔗渣提取液能更好促进灵芝菌丝生长,添加100 g/L甘蔗渣提取液,赤灵芝GL102、紫灵芝GS63和GS15菌丝生长速率分别提高48％、70％和28％.与PDA保藏和石蜡油保藏培养基相比,甘蔗渣保藏的灵芝菌丝生长快、长势好,脱氢酶活力高.因此,甘蔗渣可用于灵芝菌种的复壮和保藏.

花脸香蘑多糖具有较大的药用价值,选育高产多糖花脸香蘑菌株可丰富种质资源并促进花脸香蘑多糖科技创新和产业发展.以花脸香蘑原生质体单核体菌株LS01_10作为出发菌株,利用等离子体诱变技术选育高产胞外多糖的花脸香蘑菌株.在诱变功率为200 W,诱变时间为60 s的条件下,共获得5个性状稳定遗传的高产胞外多糖诱变菌株,其胞外多糖的产量是出发菌株的1.2-2.4倍.诱变菌株的菌丝形态和颜色都发生不同程度改变,且生物量均高于出发菌株.ISSR实验结果表明,诱变菌株与出发菌株相比在遗传水平发生不同程度的改变.高产多糖诱变菌株为进一步解析多糖高产机制提供了重要的菌种资源.

Coriolopsis trogii is a typical thermotolerant basidiomycete fungus, but its thermotolerance mechanisms are currently unknown. In this study, two monokaryons of C. trogii strain Ct001 were assembled: Ct001_29 had a genome assembly size of 38.85Mb and encoded 13,113 genes, while Ct001_31 was 40.19 Mb in length and encoded 13,309 genes. Comparative intra- and interstrain genomic analysis revealed the rich genetic diversity of C. trogii, which included more than 315,194 singlenucleotide polymorphisms (SNPs), 30,387 insertion/deletions (indels), and 1,460 structural variations. Gene family analysis showed that the expanded families of C. trogii were functionally enriched in lignocellulose degradation activities. Furthermore, a total of 14 allelic pairs of heat shock protein 20 (HSP20) genes were identified in the C. trogii genome. The expression profile obtained from RNA sequencing (RNA-Seq) showed that four tandem-duplicated allelic pairs, HSP20.5 to HSP20.8, had more than 5-fold higher expression at 35 degrees C than at 25 degrees C. In particular, HSP20.5 and HSP20.8 were the most highly expressed HSP20 genes. Allelic expression bias was found for HSP20.5 and HSP20.8; the expression of Ct29HSP20.8 was at least 1.34-fold higher than that of Ct31HSP20.8, and that of Ct31HSP20.5 was at least 1.5-fold higher than that of Ct29HSP20.5. The unique structural and expression profiles of the HSP20 genes revealed by these haplotyperesolved genomes provide insight into the molecular mechanisms of high-temperature adaptation in C. trogii. IMPORTANCE Heat stress is one of the most frequently encountered environmental stresses for most mushroom-forming fungi. Currently available fungal genomes are mostly haploid because high heterozygosity hinders diploid genome assembly. Here, two haplotype genomes of C. trogii, a thermotolerant basidiomycete, were assembled separately. A conserved tandem cluster of four HSP20 genes showing allele-specific expression was found to be closely related to high-temperature adaptation in C. trogii. The obtained haploid genomes and their comparison offer a more thorough understanding of the genetic background of C. trogii. In addition, the responses of HSP20 genes at 35 degrees C, which may contribute to the growth and survival of C. trogii at high temperatures, could inform the selection and breeding of elite strains in the future.

Hydrophobins are small, secreted proteins with important physiological functions in mycelial growth and fungal development. Here, 1 nucleus-specific and 35 allelic hydrophobin genes were identified in the genome of a white rot fungus, Coriolopsis trogii. Among these, 22 were eight-cysteine class I hydrophobin genes and the other 14 were uncommon six-cysteine hydrophobin genes. The six-cysteine hydrophobins were speculated to have originated from a common ancestor. The hydrophobin genes favored a clustering distribution and two recent duplication pairs were identified. The genes had conserved gene structures with three exons and two introns. Cthyd18, Cthyd19, and Cthyd32 were constitutively highly expressed in all developmental stages. Cthyd20, Cthyd21, Cthyd22, Cthyd28, Cthyd30, Cthyd31, and Cthyd33 were highly expressed in mycelia, and Cthyd12 and Cthyd35 in the reproductive stages. Sixteen hydrophobin genes were regulated differently in the transition from mycelia to primordia; Cthyd35 showed maximal upregulation of 1922-fold, and Cthyd23 showed maximal downregulation of 552-fold. Most (32) hydrophobin genes showed significant differential expression between mycelia cultured in different media (potato dextrose agar or broth). Weighted gene co-expression network analysis and promoter analysis revealed that C2H2 zinc finger proteins may regulate hydrophobin genes. These results may support further research into the function and evolution of hydrophobins.

为了评价肉桂木栽培食用菌的营养特性,本研究测定了采用肉桂(Cinnamomum cassia Pres1)生产中废弃物肉桂木栽培的金顶侧耳(Pleurotus citrinopileatus),秀珍菇(Pleurotus geesteranus),刺芹侧耳(Pleurotus eryngii),黑皮鸡枞(Oudemansiella raphanipes),黄伞(Pholiota adiposa)和猴头菌(Hericium erinaceus)的营养成分及氨基酸组成,通过主成分分析、蛋白质营养价值评价及聚类分析对其蛋白质的营养价值进行评价与分类.结果表明,6种肉桂栽培食用菌的膳食纤维、多糖和蛋白质含量范围分别为24％～46％、4～15 mg/g和19％～43％.主成分1(PC1)和主成分2(PC2)一共解释了92.46％的差异,苏氨酸、亮氨酸、异亮氨酸、苯丙氨酸、缬氨酸共同决定了PC1,赖氨酸决定了PC2.氨基酸评分发现,肉桂木栽培的6种食用菌中,蛋氨酸+半胱氨酸(Met+Cys)含量均过剩,与其他5种食用菌相比,秀珍菇的Met+Cys含量过剩较少,与WHO/FAO模式值相近.聚类分析结果与肉桂木栽培的食用菌的Met+Cys含量有关,黄伞与金顶侧耳距离最近,秀珍菇的Met+Cys含量与其他5种差异较大,聚类距离较远,与WHO/FAO模式值距离较近.综上表明,肉桂木栽培食用菌氨基酸含量丰富,种类比较齐全,是理想的蛋白质来源,但是由于肉桂木栽培食用菌Met+Cys含量偏高,仍需要与其他日常膳食搭配以达到人体氨基酸的平衡供给.

Numerous studies in plants have shown the vital roles of MYB transcription factors in signal transduction, developmental regulation, biotic/abiotic stress responses and secondary metabolism regulation. However, less is known about the functions of MYBs in Ganoderma. In this study, five medicinal macrofungi of genus Ganoderma were subjected to a genome-wide comparative analysis of MYB genes. A total of 75 MYB genes were identified and classified into four types: 1R-MYBs (52), 2R-MYBs (19), 3R-MYBs (2) and 4R-MYBs (2). Gene structure analysis revealed varying exon numbers (3-14) and intron lengths (7-1058 bp), and noncanonical GC-AG introns were detected in G. lucidum and G. sinense. In a phylogenetic analysis, 69 out of 75 MYB genes were clustered into 15 subgroups, and both single-copy orthologous genes and duplicated genes were identified. The promoters of the MYB genes harbored multiple cis-elements, and specific genes were co-expressed with the G. lucidum MYB genes, indicating the potential roles of these MYB genes in stress response, development and metabolism. This comprehensive and systematic study of MYB family members provides a reference and solid foundation for further functional analysis of MYB genes in Ganoderma species.

随着糙皮侧耳基因组测序的完成,基因组数据的深入挖掘和利用成为研究重点.本研究基于较高质量的糙皮侧耳CCMSSC00389-1和CCMSSC03989-1基因组图谱,开展了种内比较基因组学分析.CCMSSC00389-1和CCMSSC03989-1的11条染色体序列表现出良好的共线性,且分别含有89.92％和91.68％的保守基因.对菌株特有基因的GO富集分析表明,两菌株各自分化出一些独特的调控方式或途径.CCMSSC00389-1和CCMSSC03989-1之间共存在931 542个单核苷酸多态性位点,231 654个插入/缺失和9 221个结构变异.对与遗传变异重叠基因的GO富集分析表明,碳水化合物降解、物质运输/催化和调控/蛋白活性相关的基因分别容易发生SNP、In/Del和SV变异.CCMSSC00389-1中50.326kb缺失序列的断点两侧含有双链断裂修复(DSB)信号,推测DSB介导50.326kb序列的缺失.

Trehalose is a nonreducing disaccharide, and it plays an intracellular protective role in organisms under various stress conditions. In this study, the trehalose synthesis and its protective role in Pleurotus ostreatus were investigated. As a signal in metabolic regulation, reactive oxygen species (ROS) accumulated in the mycelia of P. ostreatus under heat stress (HS). Furthermore, mycelial growth was significantly inhibited, and the malondialdehyde (MDA) level significantly increased under HS. First, exogenous addition of H2O2 inhibited mycelial growth and elevated the MDA level, while N-acetyl cysteine (NAC) and vitamin C (VC) reduced the MDA level and recovered mycelial growth under HS by scavenging ROS. These results indicated that the mycelial radial growth defect under HS might be partly caused by ROS accumulation. Second, adding NAC and VC to the media resulted in rescued trehalose accumulation, which indicated that ROS has an effect on inducing trehalose synthesis. Third, the mycelial growth was recovered by addition of trehalose to the media after HS, and the MDA level was reduced. This effect was further verified by the overexpression of genes for trehalose-6-phosphate synthase (TPS) and neutral trehalase (NTH), which led to increased and reduced trehalose content, respectively. In addition, adding validamycin A (NTH inhibitor) to the media promoted trehalose accumulation and the recovered mycelial growth after HS. In conclusion, trehalose production was partly induced by ROS accumulation in the mycelia under HS, and the accumulated trehalose could promote the recovery of growth after HS, partly by reducing the MDA level in the mycelia.

Background Phenylalanine ammonia-lyase (PAL, EC 4.3.1.24) is the first key enzyme in the phenylpropanoid pathway. The pal gene is widely studied in plants and participates in plant growth, development and defense systems. However, the biological function of pal in P. ostreatus development and abiotic stress has not been reported. Results In this study, we cloned and characterized the pal1 (2232 bp) and pal2 (2244 bp) from basidiomycete Pleurotus ostreatus CCMSSC 00389. The pal1 and pal2 genes are interrupted by 6 and 10 introns and encode proteins of 743 and 747 amino acids, respectively. Furthermore, prokaryotic expression experiments showed that PAL enzymes catalyzed the conversion of L-phenylalanine to trans-cinnamic acid. The function of pal1 and pal2 was determined by constructing overexpression (OE) and RNA interference (RNAi) strains. The results showed that the two pal genes had similar expression patterns during different developmental stages. The expression of pal genes was higher in the reproductive growth stage than in the vegetative growth stage. And the overexpression of pal1 and pal2 resulted in the formation of primordia earlier. The results of heat stress assays showed that the RNAi-pal1 strains had enhanced mycelial tolerance to high temperature, while the RNAi-pal2 strains had enhanced mycelial resistance to H2O2. Conclusions These results indicate that two pal genes may play a similar role in the development of P. ostreatus fruiting bodies, but may alleviate stress through different regulatory pathways under heat stress.

Background: Phenylalanine ammonia-lyase (PAL, EC 4.3.1.24) is the first key enzyme in the phenylpropanoid pathway. The pal gene has been widely studied in plants and participates in plant growth, development and defense systems. However, in P leurotus ostreatus , the biological functions of pal during organismal development and exposure to abiotic stress have not been reported. Results: In this study, we cloned and characterized the pal1 (2232 bp) and pal2 (2244 bp) genes from the basidiomycete P. ostreatus CCMSSC 00389. The pal1 and pal2 genes are interrupted by 6 and 10 introns, respectively, and encode proteins of 743 and 747 amino acids, respectively. Furthermore, prokaryotic expression experiments showed that PAL enzymes catalyzed the conversion of L-phenylalanine to trans-cinnamic acid. The function of pal1 and pal2 was determined by constructing overexpression (OE) and RNA interference (RNAi) strains. The results showed that the two pal genes had similar expression patterns during different developmental stages. The expression of pal genes was higher in the reproductive growth stage than in the vegetative growth stage . And the interference of pal1 and pal2 delayed the formation of primordia. The results of heat stress assays showed that the RNAi- pal1 strains had enhanced mycelial tolerance to high temperature, while the RNAi- pal2 strains had enhanced mycelial resistance to H 2 O 2 . Conclusions: These results indicate that two pal genes may play a similar role in the development of P. ostreatus fruiting bodies, but may alleviate stress through different regulatory pathways under heat stress.

Pleurotus ostreatus is a commercially grown mushroom species in China. However, studies on the mechanisms of the fruiting body development and stress response of P. ostreatus are still at a primary stage. In this study, we report the entire genome sequence of P. ostreatus CCMSSC03989. Then, we performed comprehensive genome-wide characterization and expression analysis of the MYB transcription factor family during a series of developmental stages and under the condition of heat stress. A 34.76 Mb genome was obtained through next-generation sequencing (NGS) and Bionano optical mapping approaches. The genome has a scaffold N50 of 1.1 Mb and contains 10.11% repeats, and 10,936 gene models were predicted. A total of 20 MYB genes (PoMYB) were identified across the genome, and the full-length open reading frames were isolated. The PoMYBs were classified into 1 repeat (1R), 2R, and 3R-MYB groups according to their MYB domain repeat numbers, and 3R-MYBs possessed relatively more introns than 1R and 2R-MYBs. Based on phylogenetic analysis, the PoMYBs were divided into four groups and showed close relationships with the MYB genes of plants and fungi. RNA-sequencing (RNA-Seq) and quantitative PCR (qPCR) analyses revealed that PoMYB expression showed stage-specific patterns in reproductive stages and could be induced by heat stress. The P. ostreatus draft genome will promote genome-wide analysis, and our study of PoMYBs will promote further functional analysis of MYB genes in mushrooms.