This study employed a multi-omics approach to investigate the accumulation of bioactive terpenoids and flavonoids in Taiwanofungus gaoligongensis during liquid culture supplemented with traditional Chinese medicine materials, including Coix lacryma-jobi powder (YM), Astragalus membranaceus powder (HQ), and Pleuropterus multiflorus powder (HSW). Compared with pea flour (WDF), total terpenoid content increased significantly in the HQ and HSW groups, reaching 2.86-fold and 4.23-fold, respectively. Sesquiterpenoids were the predominant class, among which (+ /−)-α-bisabolol and nootkatone in HQ were 8.81-fold and 9.07-fold higher than those in WDF, respectively, whereas drechslerene E in HSW was elevated by 11.17-fold. Flavonoid accumulation was highest in the HQ group, reaching 16.05 times that of WDF, and was primarily composed of isoflavones. Specifically, the levels of maximaisoflavone J, glycitein 1, and formononetin in HQ were 56.59-, 460.39-, and 4570.37-fold higher than those in WDF, respectively. Moreover, the characteristic active component astragaloside IV from A. membranaceus and the flavonoids quercetin and rutin from P. multiflorus were detected in the HQ and HSW fermentation products, respectively. In contrast, potentially hepatotoxic constituents of P. multiflorus, including emodin and anthraquinones, were not detected in the HSW fermentation products, suggesting that co-fermentation with T. gaoligongensis may attenuate the toxicity of P. multiflorus. Transcriptomic analysis revealed that the relative expression levels of multiple genes involved in terpenoid biosynthesis were significantly upregulated in the HQ and HSW groups. In HSW, the expression levels of TgHMGS, TgHMGR, TgMVD, and TgOSC were 2.68-, 2.62-, 2.30-, and 1.86-fold higher than those in WDF, respectively, whereas TgAACT, TgIDI, TgSES, and TgErg25 were upregulated by 2.11-, 1.82-, 2.68-, and 2.25-fold in HQ. These genes may represent key regulatory nodes in the terpenoid biosynthetic pathway of T. gaoligongensis. Furthermore, co-expression trend analysis combined with promoter-binding site prediction suggested that TgZnF10, TgHMG11, and TgZnF19 may regulate the transcriptional activity of TgHMGR, TgPMK, and TgFPPS_2, respectively, while TgErg26, TgErg62, TgErg4, and TgErg5 may be regulated by TgBHLH7, TgBZIP4, TgMYB3, and TgHOX2, respectively. Collectively, these findings provide a theoretical basis for the comprehensive utilization and industrial development of T. gaoligongensis through bidirectional fermentation with Chinese medicinal materials.

Slope aspect and slope position have an important influence on plant growth by changing the microclimate and soil conditions such as light, temperature, moisture, and nutrients. In this study, 15-year-old Sapindus mukorossi forests with different slope aspects and positions were selected and the differences in tree height and diameter at breast height (DBH), leaf non-structural carbohydrate (NSC) characteristics, and leaf–soil nitrogen (N), phosphorus (P), and potassium (K) stoichiometric characteristics between sunny and shady slopes, and upper, middle, and down slope positions were compared and analyzed. The results show that the tree height and DBH of S. mukorossi were better in the same slope aspect and lower slope position, while in the same slope position, the tree height and diameter at DBH were better on the shady slopes. In the upper slope position, the starch content on the shady slope was significantly higher than that on the sunny slope, and the NSC content was significantly higher than that on the sunny slope. On shady and sunny slopes, S. mukorossi is mainly limited by N. The leaf and soil P content of S. mukorossi on the sunny slope was the highest and significantly higher than that on the upper slope. The coefficient of variation of each index of S. mukorossi on the shady slope and the sunny slope was medium and below. Soil N/P, soil N, soil N/K, soluble sugar/starch, leaf P, leaf K, leaf N, and soil K had strong plasticity under different slope aspects. Therefore, it indicated that the shady slope and down slope were more suitable for S. mukorossi.

Phlebopus portentosus is an edible and medicinal ectomycorrhizal mushroom with delicious and high nutritional value. However, the mechanism of secondary metabolite biosynthesis in P. portentosus is still unclear. In this study, the genomics, metabolomics, and transcriptomics were integrated to reveal the biosynthesis mechanism of secondary metabolites in P. portentosus under different cultivation conditions. The 31.4 Mb genome of P. portentosus YAF023 with 15 scaffolds was assembled by Illumina and Nanopore sequencing and annotated, and 206 cytochrome P450s, 201 carbohydrate-active enzymes, 186 transcription factors, 18 terpene synthases (TPSs), and 5 polyketide synthases (PKSs) were identified. Multi-omics analysis showed that PpPKS1 is probably involved in the biosynthesis of Ethyl orsellinate; PpPKS2 and PpPKS5 are probably involved in the synthesis of 6-Methylsalicylic acid and Cytochalasin Z5, respectively; PpTRI5 was involved in the tetracyclic sesquiterpene β-type trichodiene compounds; and PpSTCs was involved in the synthesis of β-copaene analogs or derivatives. Co-expression network analysis and binding site prediction of the promoter regions suggested that PpHOX4 and PpHSF1 regulated the gene expression of PpPKS1, and Ppzf-C2H2 32 and PpHSF5 regulated the gene expression of PpSTCs 8, and PpSTCs 3, respectively. This study will provide an important foundation for further development and utilization of secondary metabolites of P. portentosus.

Introduction:The fungal genus Inonotus is renowned for its medicinal properties, including antioxidant and anti-tumor activities, which are largely attributed to its rich repertoire of terpenoid and polyphenolic secondary metabolites. This study aimed to investigate how different Chinese herbal medicine powders used as culture media influence the secondary metabolite profile of Inonotus glomeratus. Methods:This study employed a multi-omics approach, utilizing Illumina and Nanopore sequencing to assemble a high-quality genome for I. glomeratus. The fungus was cultivated on media containing powders from Polygonum multiflorum, Coix lacryma-jobi, Pisum sativum flour, Salvia miltiorrhiza, Panax ginseng, and Astragalus membranaceus. Subsequent integrated metabolomic and transcriptomic analyses were conducted to profile secondary metabolite production and identify key biosynthetic genes. Results:Experimental results show that the assembled I. glomeratus genome was 38.68 Mb in size, consisting of 23 scaffolds with a GC content of 47.98%. The genome annotation process identified 67 transcription factors, four polyketide synthases (PKSs), one non-ribosomal peptide synthase, and 11 terpenoid synthases (TPSs). Multi-omics analysis revealed that terpenoid biosynthesis in I. glomeratus was significantly enhanced in DS and RS media. Betulin and betulinic acid exhibited the most dramatic increases in RS medium, reaching 4,658-9,275-fold and 4-503-fold higher concentrations, respectively. The transcriptome results showed that the expression of enzymes such as IgAACT, IgHMGR, IgSQS, IgSES, and IgTPS9 was significantly higher in the RS medium than in the other treatment groups. Integrated metabolomic and transcriptomic analyses suggested that IgPKS1 participates in orsellinic acid biosynthesis. while IgPKS2 is likely involved in naringenin biosynthesis. Additionally, IgTPS9 was associated with betulinic acid biosynthesis, and IgTPS10 contributed to tetracyclic sesquiterpene B-type triterpene formation. Co-expression network analysis and transcription factor binding site prediction indicated that IgMYB3 may regulates IgPKS1 expression, whereas IgHSF1 may simultaneously modulate IgTPS4 and IgTPS6. Discussion:This study provides novel insights into the regulatory mechanisms governing secondary metabolite production in I. glomeratus. These findings offer a foundation for targeted metabolic engineering and the optimized production of valuable compounds.

A multi-omics strategy was utilized in this study to investigate the effects of various cultivation methods-including the fruiting bodies cultivation on Cinnamomum kanehirae wood logs (GLG), the mycelia cultivation on C. kanehirae substrate fungal cultivation bags (NZJB), Cinnamomum camphora substrate fungal cultivation bags (XZJB) and rice medium (DM)-on Secondary Metabolites in Taiwanofungus gaoligongensis. NZJB and XZJB significantly enhanced terpenoids production in the mycelium, with triterpenoid contents in NZJB and XZJB being sevenfold and 3.9-fold higher, respectively, than those in DM. Antcins were notably increased in fungal cultivation bag cultures: antcin C reached the highest level in XZJB (9.72-fold higher than in DM), antcin I peaked in NZJB (12.83-fold higher than in DM), and antrodin C also reached its maximum in NZJB. Additionally, the antrodin C content in NZJB was 3.2-fold higher than in GLG and 4.08-fold higher than in DM. In addition, the levels of steroids, phenolic compounds, and flavonoids were also significantly increased in NZJB and XZJB. Transcriptome analysis revealed significant differences in the expression of genes involved in the biosynthesis of antcins and antrodin C across the different cultivation methods. In particular, the expression of TgHMGR was markedly higher in NZJB than in XZJB and DM, correlating with the elevated terpenoids and triterpenoids levels, suggesting that TgHMGR may act as a key rate-limiting enzyme in the terpenoid biosynthesis pathway of T. gaoligongensis. The expression levels of terpenoid biosynthesis-related genes were significantly elevated in GLG compared to mycelium, consistent with the higher abundance of terpenoid metabolites. Co-expression analysis of transcription factors (TFs) and promoter binding site predictions indicated that the expression of TgHMGR and TgFPPS 2 may be regulated by TgHSF4 and TgMYB6, respectively. Meanwhile, the expression of TgErg2, TgErg3, TgErg5, and TgErg6 1 may be regulated by TgZnF1, TgMYB9, TgHOX1, and TgHMG8. This study compared the metabolite profiles and gene expression patterns of the fruiting bodies of T. gaoligongensis with those of three types of cultivated mycelia. The results provide new insights into the transcriptional regulation of key bioactive compound biosynthesis in T. gaoligongensis and suggest potential strategies to enhance the production of active compounds in mycelia through artificial cultivation, thereby improving its medicinal value and production efficiency.

Salicylic acid (SA) serves as an intercellular signaling molecule, playing a crucial role in plant growth and development, along with the response to environmental stressors. However, molecular regulations that govern salicylic acid-induced resistance to drought in plants remain incompletely elucidated. This research utilized two-year-old C. camphora seedlings as the experimental subjects, employing a two-factor experimental design that incorporated soil moisture×salicylic acid spraying. Through a combination of physiological and transcriptomic analyses, it aimed to elucidate the mechanisms by which exogenous salicylic acid influences the growth and physiological traits of C. camphora seedlings subjected to drought stress, as well as the regulation of salicylic acid-mediated drought-related signaling pathways. Research indicates that SA can markedly improve the substance called chlorophyll fluorescence parameters (that is, Fv/Fm and PIabs) of C. camphora subjected to drought stress, augment photosystem activity during mild drought conditions, and mitigate the damage inflicted by excessive light energy in photosynthetic institutions. SA significantly alleviated oxidative stress in C. camphora seedlings under drought stress by reducing O2- and H2O2 contents and enhancing SOD, POD, and CAT activities. Transcriptome analysis revealed that SA induces DEGs associated with drought resistance. It activates transcription factors that are attached as NAC, bHLH, ERF, and MYB, and regulates genes involved in plant hormone signaling, such as AUX/IAA, PYR/PYL, A-ARRs, and B-ARRs. Additionally, it suppresses the degradation of starch, enhances the expression of genes associated with photosynthesis, and alleviates the adverse effects during conditions of drought that negatively impact the photosynthetic performance of C. camphora, thus enhancing their resilience to drought conditions. Furthermore, SA significantly affected phenylpropanoid synthesis-related genes (such as CcHCT, CcPOD, and CcCOMT). This research seeks to improve understanding of the mechanisms by which SA influences drought tolerance in plants, providing novel insights into enhancing drought resistance in C. camphora.

The Amyloporia genus is an important Chinese medicinal fungus. Here, we present a draft genome sequence of Amyloporia xantha strain YAFMF0618. The genome resource will support subsequent research into the potential secondary metabolite diversity of A. xantha.

Fungal secondary metabolites (SMs) have broad applications in biomedicine, biocontrol, and the food industry. In this study, whole-genome sequencing and annotation of Taiwanofungus gaoligongensis were conducted, followed by comparative genomic analysis with 11 other species of Polyporales to examine genomic variations and secondary metabolite biosynthesis pathways. Additionally, transcriptome data were used to analyze the differential expression of polyketide synthase (PKS), terpene synthase (TPS) genes, and transcription factors (TFs) under different culture conditions. The results show that T. gaoligongensis differs from other fungal species in genome size (34.58 Mb) and GC content (50.72%). The antibiotics and Secondary Metabolites Analysis Shell (AntiSMASH) analysis reveals significant variation in the number of SM biosynthetic gene clusters (SMBGCs) across the 12 species (12–29), with T. gaoligongensis containing 25 SMBGCs: 4 PKS, 6 non-ribosomal peptide synthetase (NRPS), and 15 TPS clusters. The TgPKS1 gene is hypothesized to be involved in the biosynthesis of orsellinic acid or its derivatives, while TgPKS2 might catalyze the synthesis of 6-methylsalicylic acid (6MSA) and its derivatives. The TgTRI5 genes are suggested to synthesize tetracyclic sesquiterpene type B trichothecene compounds, while TgPentS may be involved in the synthesis of δ-cadinol, β-copaene, and α-murolene analogs or derivatives. Comparative genomic analysis shows that the genome size of T. gaoligongensis is similar to that of T. camphoratus, with comparable SMs. Both species share four types of PKS domains and five distinct types of TPS. Additionally, T. gaoligongensis exhibits a high degree of similarity to Laetiporus sulphureus, despite belonging to a different genus within the same family. Transcriptome analysis reveals significant variation in the expression levels of PKS and TPS genes across different cultivation conditions. The TgPKS1 and TgPKS4 genes, along with nine TgTFs, are significantly upregulated under three solid culture conditions. In contrast, under three different liquid culture conditions, the TgPKS3, TgTRI5-1, and TgTRI5-2 genes, along with twelve TgTFs, exhibit higher activity. Co-expression network analysis and TgTFs binding site prediction in the promoter regions of TgPKS and TgTPS genes suggest that TgMYB9 and TgFTD4 regulate TgPKS4 expression. TgHOX1, TgHSF2, TgHSF3, and TgZnF4 likely modulate TgPKS3 transcriptional activity. TgTRI5-1 and TgTRI5-5 expression is likely regulated by TgbZIP2 and TgZnF15, respectively. This study provides new insights into the regulatory mechanisms of SMs in T. gaoligongensis and offers potential strategies for enhancing the biosynthesis of target compounds through artificial intervention.

The draft genome sequence of Taiwanofungus gaoligongensis YAF008 was reported. The genome size of T. gaoligongensis YAF008 was 34.7M bp with 50.72% GC content. The genome resource will support future research into potential secondary metabolite diversity of this fungus.

Persea americana Mill. is an important cash crop that contains effective ingredients to reduce cholesterol and protect the cardiovascular system. Presently, the gene regulation mechanism and signal pathway of stress response in P. americana are unclear. To explore the gene expression changes of P. americana under drought and low-temperature stress, the transcripts of P. americana were sequenced under these conditions. The results produced 42,815,960 bp raw reads. Analysis of the related metabolic pathways and differentially expressed genes showed that under drought stress, the gene expression of beta-amylase 3, glyceraldehyde-3-phosphate dehydrogenase and hexokinase were upregulated, while the gene expression of UDP-glycosyltransferase superfamily protein isoform, glucose-1-phosphate adenylyltransferase, and glucose-6-phosphate 1-epimerase were downregulated. Under low-temperature stress, the expression of beta-amylase and shikimate O-hydroxycinnamoyl transferase genes was downregulated. In addition, WRKY, MYB, bHLH, and NAC transcription factors were expressed under drought and low-temperature stress. Finally, the RNA-Seq data were validated using real-time fluorescence quantitative analysis to identify the key genes of P. americana regulated at the transcriptional level under drought and low-temperature stress. This study provides a theoretical basis for the selection of drought-resistant and low-temperature tolerant P. americana varieties.

Antimicrobial resistance is a major threat to human health globally. Antrodia camphorata was grown in a malt/yeast extract broth liquid medium for 15 days. Then, 4-L fermentation broth was harvested, yielding 7.13 g of the ethyl acetate extract. By tracing the antimicrobial activity, 12.22 mg of the antimicrobial compound was isolated. The structure of 5-methyl-benzo [1,3]-dioxole-4,7-diol (MBBD) was elucidated using NMR and MS data analyses. The antibacterial activity of MBBD was detected through the microbroth dilution method. MBBD exhibited broad-spectrum antibacterial activity. The minimum inhibitory concentration (MIC) range of MBBD for drug-resistant pathogenic bacteria was 64-256 μg/mL, with the lowest MIC observed for Acinetobacter baumannii (64 μg/mL), followed by Pseudomonas aeruginosa (MIC = 128 μg/mL). Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli were also sensitive, with an MIC of 256 μg/mL. The MIC range of MBBD against 10 foodborne pathogens was 12.5-100 μg/mL. Based on the results of this study, MBBD exhibits broad-spectrum antibacterial activity, particularly demonstrating excellent inhibitory effects against A. baumannii. MBBD will be good candidates for new antimicrobial drugs.

Antrodia cinnamomea, an edible and medicinal fungus with significant economic value and application prospects, is rich in terpenoids, benzenoids, lignans, polysaccharides, and benzoquinone, succinic and maleic derivatives. In this study, the transcriptome of A. cinnamomea cultured on the wood substrates of Cinnamomum glanduliferum (YZM), C. camphora (XZM), and C. kanehirae (NZM) was sequenced using the high-throughput sequencing technology Illumina HiSeq 2000, and the data were assembled by de novo strategy to obtain 78,729 Unigenes with an N50 of 4,463 bp. Compared with public databases, about 11,435, 6,947, and 5,994 Unigenes were annotated to the Non-Redundant (NR), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG), respectively. The comprehensive analysis of the mycelium terpene biosynthesis-related genes in A. cinnamomea revealed that the expression of acetyl-CoA acetyltransferase (AACT), acyl-CoA dehydrogenase (MCAD), 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA), mevalonate pyrophosphate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI) was significantly higher on NZM compared to the other two wood substrates. Similarly, the expression of geranylgeranyltransferase (GGT) was significantly higher on YZM compared to NZM and XZM, and the expression of farnesyl transferase (FTase) was significantly higher on XZM. Furthermore, the expressions of 2,3-oxidized squalene cyclase (OCS), squalene synthase (SQS), and squalene epoxidase (SE) were significantly higher on NZM. Overall, this study provides a potential approach to explore the molecular regulation mechanism of terpenoid biosynthesis in A. cinnamomea.

Abstract Antrodia cinnamomea is a rare medicinal fungus endemic to Taiwan, China. It has a high medicinal value, being rich in bioactive components such as triterpenes and polysaccharides. However, only a few studies have been conducted on A. cinnamomea fruiting bodies grown on Cinnamomum camphora wood culture. In this study, we performed transcriptome sequencing of A. cinnamomea fruiting bodies grown on C. kanehirae (NZF) and C. camphora (XZF) wood substrates. 18391 unigenes were annotated from the obtained reads, including 592 differentially expressed genes (170 up-regulated and 422 down-regulated) mainly enriched in the carbohydrate metabolism, lipid metabolism, and amino acid metabolism pathways. Comparing the transcriptome of NZF and XZF, genes encoding acetyl CoA synthetase and acetyl CoA C-acetyltransferase, involved in the terpenoid backbone biosynthesis, were highly expressed in NZF, while genes encoding glycosyltransferases and cytochrome P450s were highly expressed in XZF. There were no significant differences in the expression of the genes encoding farnesyl diphosphate farnesyltransferase, isopentenyldiphosphate isomerase, 2,3-oxidosqualene cyclase, and terpenoid synthase between NZF and XZF. The transcriptome analysis further supports that C. camphora is feasible as an alternative host of A. cinnamomea.

MYB转录因子在真菌的生长发育和胁迫响应中发挥多种作用.为了解牛樟芝MYB的功能,本研究利用本地BLAST对牛樟芝基因组进行比对分析,获得了 9个牛樟芝MYB基因序列,并对牛樟芝MYB转录因子进行功能预测分析,利用转录组测序分析MYB基因在不同段木培养条件下子实体和菌丝体中的表达情况.蛋白理化性质分析发现,9个牛樟芝MYB转录因子多为不稳定蛋白,理论等电点介于4.92～8.51;亚细胞定位预测结果显示有1个转录因子(ACMYB1)定位在细胞质中,推测其可能参与细胞质基因的转录调控;有8个定位在细胞核中,占总蛋白的88.9％.二级结构预测发现以无规则卷曲和a-螺旋为主要结构,β-转角为次要结构.将9个牛樟芝MYB转录因子与5个已知功能的MYB蛋白共同构建系统进化树,14个MYB转录因子分为Ⅰ、Ⅱ两个分支,其中ACMYB1、ACMYB5与冬虫夏草中参与菌丝生长的MYB-6聚为一支,ACMYB9与冬虫夏草中参与子实体生长的MYB-3聚为一支,ACMYB6与顶头孢霉AcMYBA聚为一支.分析牛樟芝MYB转录因子在不同段木菌丝和子实体中的相对表达量,发现ACMYB1、ACMYB5、ACMYB6均在香樟段木菌丝体阶段高表达,推测其参与了牛樟芝菌丝体的生长,ACMYB9基因在香樟段木子实体阶段表达量上调,说明ACMYB9可能参与调控牛樟芝子实体的发育.本研究结果可为进一步挖掘研究牛樟芝MYB转录因子的功能奠定基础.

Objective Effects and optimal amounts of branches and/or leaves of Cinnamomum bodinieri added in culture substrate on the growth and physiochemical characteristics of Antrodia cinnamomea were determined.Method Dish culture method was used for the experimentation. Biomass of the cultivated mushrooms was determined by drying method, total triterpenes (TT) content in mycelium by vanillin-glacial acetic acid method, and superoxide dismutase (SOD) activity by nitrogen blue tetrazolium photoreduction method. ResultWhen various concentrations of C. bodinieri young branches, i.e., 0.5-4 g·L−1, 2-8 g·L−1, 1-4 g·L−1, and 4-16 g·L−1, were added to the PDA medium, the growth, biomass, SOD activity, and TT content of A. cinnamomea were significantly higher than those of control without the addition. With 1-2 g·L−1 added leaves in the substrate, A. cinnamomea grew well with an increased SOD activity; while at 2-4 g·L−1, raised biomass and TT content. By adding both branches and leaves at 2-4 g·L−1, 1-4 g·L−1, 0.125-1 g·L−1, or 4-8 g·L−1, the mushroom growth could be improved with increases on biomass, SOD activity, and TT content. Conclusion Adding 2 g·L−1 of C. bodinieri leaves to PDA medium for the mushroom cultivation resulted in the most significant effect on the growth and biomass gain of A. cinnamomea mycelia. At the addition level of 1 g·L−1 or 2 g·L−1, the greatest enhancement on the mycelial SOD activity was observed. With an 8 g·L−1 branches addition, the TT content of the mycelia significantly increased to 23.73 mg·g−1, representing a rise of 81.77% over control.

WRKY转录因子是植物逆境研究中最广泛的家族之一,在植物响应生物胁迫和非生物胁迫中具有重要作用.本研究利用本地BLAST对牛樟基因组进行比对分析得到58个牛樟WRKY基因序列,并对牛樟WRKY转录因子进行理化性质、保守基序、系统进化及表达谱分析.保守结构域和基序预测发现,牛樟转录因子家族蛋白主要分为三大类,即Ⅰ、Ⅱ、Ⅲ类,多数蛋白较保守,其中Ⅰ、Ⅱ类中有两个蛋白(CkWRKY25,CkWRKY1)存在Q残基的变异;CkWRKY基因表达谱数据分析发现,有22个基因在受干旱、低温胁迫时上调表达,占牛樟WRKY家族的37.9％,其中18个基因受干旱胁迫呈高表达水平,占上调基因的81.8％,4个基因在4℃低温胁迫下表达量显著升高,占比18.2％.将牛樟中22个表达差异显著的基因与14个具有抗旱、抗寒WRKY基因共同建树,结果显示36个WRKYs蛋白聚为两大类,参与干旱及低温胁迫的蛋白主要集中在第一大类中,其中CkWRKY26、CkWRKY38、CkWRKY39、CkWRKY45、CkWRKY55分别和参与干旱胁迫的蛋白聚为一支,且C端锌指结构均为C2H2;CkWRKY20、CkWRKY54和参与低温胁迫中陆地棉的GhWRKY62、毛果杨的PtWRKY34、西洋参的PqWRKY7、旱地棉的GaWRKY28、陆地棉的GhWRKY73以及小麦的TaWRKY19和TaWRKY10聚为一支.本研究基于牛樟高通量转录组数据,探究牛樟WRKY转录因子家族生物学特性,为深入研究牛樟WRKY基因非生物胁迫调控提供科学依据.

NAC蛋白是最大的植物特异性转录因子家族之一,在植物响应生物胁迫和非生物胁迫中具有重要作用.本研究利用二代转录组测序获得牛樟干旱及低温胁迫的转录组数据,并利用本土BLAST从基因组数据中获得牛樟全基因组78个CkNAC转录因子序列,利用分析软件进行保守基序分析.利用转录组分析获得与干旱和低温胁迫相关的CkNAC,进行系统进化和相关基因荧光定量PCR验证.牛樟NAC基因表达谱数据分析发现,有18个基因在受干旱、低温胁迫时表达差异显著,占牛樟NAC家族的23.1％,其中10个基因受干旱胁迫呈高表达水平,占上调基因的55.6％.将14个牛樟NAC转录因子与8个具有抗旱、抗寒NAC蛋白共同构建系统发育进化树,结果显示22个NACs蛋白聚为Ⅰ类和Ⅱ类,参与干旱和低温胁迫的蛋白集中在Ⅰ类.Ⅰ类中CkNAC14、CkNAC23、CkNAC44和CkNAC70分别和参与干旱胁迫的普通小麦(Triticum aestivum)TaNAC67、梭梭(Haloxylon ammodendron)HaNAC38、拟南芥(Arabidopsis thaliana)ANAC072和大豆(Glycine max)GmNAC20聚为一支,其中CkNAC44和CkNAC70与ANAC072和GmNAC20的相似性分别为59.0％ 和58.2％,推测CkNAC14、CkNAC23、CkNAC44和CkNAC70可能参与调控牛樟在干旱逆境下的反应.利用RT?qPCR技术检测牛樟NAC基因在不同逆境胁迫中的表达变化,结果显示,CkNAC14、CkNAC23、CkNAC44和CkNAC70受干旱胁迫诱导显著上调.本研究基于牛樟高通量转录组数据,探究牛樟NAC转录因子家族生物学特性,为深入研究牛樟NAC基因的非生物胁迫调控提供科学依据.

Stout camphor tree (Cinnamomum kanehirae) is an economically and socially valuable timber species that is conserved in Taiwan Province, China. In this study, Illumina sequencing technology was used to identify genes in C. kanehirae and analyze their levels under drought and low temperature stresses. The obtained reads generated 27,885 single genes, including 8,136 that were differential. Based on the results of Kyoto Encyclopedia of Genes and Genomes enrichment, the primary metabolic pathways in response to drought and low temperature stress included sucrose and starch metabolism, photosynthesis, glycolysis and sugar metabolism synthesis, phenylpropanoid biosynthesis, plant-pathogen interaction, and flavonoid biosynthesis, as well as the identification of transcription factors (TFs) with different patterns of expression. Finally, the RNA-Seq data were validated using real-time fluorescence quantitative analysis to identify TFs with different patterns of expression. These data provide a valuable resource for further research on the molecular mechanisms of drought and low temperature stress in C. kanehirae and contribute to the exploration of drought and cold resistance genes.

The rare edible and medicinal fungus Antrodia cinnamomea has a substantial potential for development. In this study, Illumina HiSeq 2000 was used to sequence its transcriptome. The results were assembled de novo, and 66,589 unigenes with an N50 of 4413 bp were obtained. Compared with public databases, 6,061, 3,257, and 2,807 unigenes were annotated to the Non-Redundant, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes databases, respectively. The genes related to terpene biosynthesis in the mycelia of A. cinnamomea were analyzed, and acetyl CoA synthase (ACS2 and ACS4), hydroxymethylglutaryl CoA reductase (HMGR), farnesyl transferase (FTase), and squalene synthase (SQS) were found to be upregulated in XZJ (twig of C. camphora) and NZJ (twig of C. kanehirae). Moreover, ACS5 and 2,3-oxidized squalene cyclase (OCS) were highly expressed in NZJ, while heme IX farnesyl transferase (IX-FIT) and ACS3 were significantly expressed in XZJ. The differential expression of ACS1, ACS2, HMGR, IX-FIT, SQS, and OCS was confirmed by real-time quantitative reverse transcription PCR. This study provides a new concept for the additional exploration of the molecular regulatory mechanism of terpenoid biosynthesis and data for the biotechnology of terpenoid production.

[目的]明确添加猴樟Cinnamomum bodinieri基质对牛樟芝Antrodia cinnamomea生长及生理生化特性的影响,并筛选出适宜的猴樟茎、叶浓度.[方法]比较分析在牛樟芝基础培养基中添加质量浓度分别为0.125、0.25、0.5、1、2、4、8、16、32、64 g·L?1的猴樟嫩枝、嫩叶、枝叶混合物时,牛樟芝的生长特性、生物量、SOD活性和总三萜(TT)含量的差异.[结果]在PDA培养基中添加质量浓度分别为0.5～4 g·L?1、2～8 g·L?1、1～4 g·L?1、4～16 g·L?1的猴樟嫩枝基质,牛樟芝相应表现出生长特性较优、生物量、SOD活性和TT含量较高的优势,并显著高于对照水平;在PDA培养基中添加猴樟嫩叶基质质量浓度为1～2 g·L?1时,其生长特性较优,SOD活性较高,当添加量为2～4 g·L?1时,其生物量和TT含量较高;在PDA培养基中添加猴樟枝叶混合物基质质量浓度分别为2～4 g·L?1、1～4 g·L?1、0.125～1 g·L?1、4～8 g·L?1时,其相应表现为生长特性较优、生物量、SOD活性和TT含量较高的优势.[结论]研究结果表明,总体来说,在PDA培养基中添加猴樟基质均能促进牛樟芝菌丝体、生物量、SOD活性和总三萜含量的提高,其中猴樟嫩叶基质2 g·L?1对菌丝体和生物量的提高最显著、猴樟嫩枝1 g·L?1或嫩叶基质2 g·L?1对菌丝体SOD活性的促进效果最显著;猴樟嫩枝基质8 g·L?1,对菌丝体总三萜含量的促进效果最显著,达到23.73 mg·g?1,较对照组提高了81.77％.该结果是樟属植物对牛樟芝培养的补充,为牛樟芝的规模化生产和开发利用提供了理论依据.