ganoderic-acid and methyl-jasmonate

Ganoderma lucidum, which contains numerous biologically active compounds, is known worldwide as a medicinal basidiomycete. Because of its application for the prevention and treatment of various diseases, most of artificially cultivated G. lucidum is output to many countries as food, tea, and dietary supplements for further processing. Methyl jasmonate (MeJA) has been reported as a compound that can induce ganoderic acid (GA) biosynthesis, an important secondary metabolite of G. lucidum. Herein, MeJA was found to increase the intracellular level of nitric oxide (NO). In addition, upregulation of GA biosynthesis in the presence of MeJA was abolished when NO was depleted from the culture. This result demonstrated that MeJA-regulated GA biosynthesis might occur via NO signaling. To elucidate the underlying mechanism, we used gene-silenced strains of nitrate reductase (NR) and the inhibitor of NR to illustrate the role of NO in MeJA induction. The results indicated that the increase in GA biosynthesis induced by MeJA was activated by NR-generated NO. Furthermore, the findings indicated that the reduction of NO could induce GA levels in the control group, but NO could also activate GA biosynthesis upon MeJA treatment. Further results indicated that NR silencing reversed the increased enzymatic activity of NOX to generate ROS due to MeJA induction. Importantly, our results highlight the NR-generated NO functions in signaling crosstalk between reactive oxygen species and MeJA. These results provide a good opportunity to determine the potential pathway linking NO to the ROS signaling pathway in fungi treated with MeJA. KEY POINTS: • MeJA increased the intracellular level of nitric oxide (NO) in G. lucidum. • The increase in GA biosynthesis induced by MeJA is activated by NR-generated NO. • NO acts as a signaling molecule between reactive oxygen species (ROS) and MeJA.

Topics: Acetates; Cyclopentanes; Nitrate Reductase; Nitric Oxide; Oxylipins; Reishi; Triterpenes

Topics: Acetates; Chromatography, High Pressure Liquid; Cyclopentanes; Energy Metabolism; Gas Chromatography-Mass Spectrometry; Mass Spectrometry; Metabolome; Metabolomics; Oxylipins; Proteome; Proteomics; Reishi; Triterpenes

Heat stress (HS) is an important environmental factor that affects the growth and metabolism of edible fungi, but the molecular mechanism of the heat stress response (HSR) remains unclear. We previously reported that HS treatment increased the length between two hyphal branches and induced the accumulation of ganoderic acid biosynthesis and the gene expression of heat shock proteins (HSPs) in Ganoderma lucidum. In this study, we found that HS induced a significant increase in the cytosolic ROS concentration, and exogenously added ROS scavengers NAC, VC and NADPH oxidase (Nox) inhibitor DPI reduce the cytosolic ROS accumulation in G. lucidum. In addition, the phenomena of the increased gene expression and increased length between the two hyphal branches and the accumulation of GA biosynthesis induced by HS were mitigated. Furthermore, we investigated the effects of HS on Nox-silenced strains (NoxABi-10, NoxABi-11 and NoxRi-4, NoxRi-7) and found that the level of ROS concentration was lower than that in wild-type (WT) strains treated with HS. Additionally, Nox silenced strains reduced the HS-induced increase in HSP expression, the length between two hyphal branches and GA biosynthesis compared with the WT strain. These data indicate that HS-induced ROS participate in the regulation of HSP expression, hyphal branching and ganoderic acid biosynthesis in G. lucidum. In addition, these findings identified potential pathways linking ROS networks to HSR, physiological and metabolic processes in fungi and provide a valuable reference for studying the role of ROS in HSR, mycelium growth and secondary metabolites.

Topics: Acetates; Antioxidants; Cyclopentanes; Heat-Shock Proteins; Heat-Shock Response; Hydrogen Peroxide; Hyphae; NADPH Oxidases; Oxidation-Reduction; Oxidative Stress; Oxylipins; Reishi; Triterpenes

Ganoderma lucidum is a mushroom with traditional medicinal properties that has been widely used in China and other countries in Eastern Asia. Ganoderic acids (GA) produced by G. lucidum exhibit important pharmacological activities. Previous studies have demonstrated that methyl jasmonate (MeJA) is a potent inducer of GA biosynthesis and the expression of genes involved in the GA biosynthesis pathway in G. lucidum. To further explore the mechanism of GA biosynthesis, cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) was used to identify genes that are differentially expressed in response to MeJA. Using 64 primer combinations, over 3910 transcriptionally derived fragments (TDFs) were obtained. Reliable sequence data were obtained for 390 of 458 selected TDFs. Ninety of these TDFs were annotated with known functions through BLASTX searching the GenBank database, and 12 annotated TDFs were assigned into secondary metabolic pathways by searching the KEGGPATHWAY database. Twenty-five TDFs were selected for qRT-PCR analysis to confirm the expression patterns observed with cDNA-AFLP. The qRT-PCR results were consistent with the altered patterns of gene expression revealed by the cDNA-AFLP technique. Additionally, the transcript levels of 10 genes were measured at the mycelium, primordia, and fruiting body developmental stages of G. lucidum. The greatest expression levels were reached during primordia for all of the genes except cytochrome b2 reached its highest expression level in the mycelium stage. This study not only identifies new candidate genes involved in the regulation of GA biosynthesis but also provides further insight into MeJA-induced gene expression and secondary metabolic response in G. lucidum.

Topics: Acetates; Amplified Fragment Length Polymorphism Analysis; Base Sequence; Cyclopentanes; DNA, Complementary; Gene Expression Profiling; Gene Expression Regulation, Developmental; Gene Expression Regulation, Plant; Genes, Plant; Metabolic Networks and Pathways; Oxylipins; Reishi; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription, Genetic; Triterpenes