Ganoderma lucidum is a mushroom widely used for its edible and medicinal properties. Primary bioactive constituents of G. lucidum are ganoderic triterpenoids (GTs), which exhibit important pharmacological activity. Abscisic acid (ABA), a plant hormone, is associated with plant growth, development, and stress responses. ABA can also affect the growth, metabolism, and physiological activities of different fungi and participates in the regulation of the tetracyclic triterpenes of some plants. Our findings indicated that ABA treatment promoted GT accumulation by regulating the gene expression levels (squalene synthase (sqs), 3-hydroxy-3-methylglutaryl-CoA reductase (hmgr), and lanosterol synthase (ls)), and also activated cytosolic Ca²⁺ channels. Furthermore, under ABA mediation, exogenous Ca²⁺ donors and inhibitors directly affected the cytosolic Ca²⁺ concentration and related gene expression in Ca²⁺ signaling. Our study also revealed that ABA-mediated cytosolic Ca²⁺ played a crucial regulatory role in GT biosynthesis, accompanied by antioxidant defense modulation with increasing superoxide dismutase (SOD) activity and ascorbate peroxidase (APX) activity, and the resistance ability of O₂^•- and glutathione (GSH) contents.
Reishi/metabolism* ; Triterpenes/metabolism* ; Abscisic Acid/metabolism* ; Antioxidants/metabolism*

To study the effect and mechanism of the light quality acting on Ganoderma lucidum, and provide a theoretical basis for G. lucidum mycelium cultivation, we focused on growth and endogenous IAA metabolism of G. lucidum mycelium under different light-emitting diode (LED) condition. The growth index, endogenous levels of IAA and Enzymes related to IAA metabolism and Polysaccharides content were investigated in different growth periods. Results showed that blue light irradiation was the best from the viewpoint of steady growth and polysaccharides accumulation, red light irradiation improved endogenous IAA level and promoted growth of mycelium in early stage of cultivation, green light irradiation decreased growth rate and fresh weight of mycelium, but increased drying rate. Enzymes related to IAA metabolism also significantly influenced by light quality. The activity of indole acetic acid oxidase (IAAO), peroxidase (POD) and tryptophan synthetase with blue light irradiation were showed high level in early time, but decreased later, and the IAA content was consistently at lower level than that in other treatments, while mycelium irradiated with yellow light showed the highest activity of both IAAO and tryptophan synthetase, and medium level of IAA content. In conclusion, the light quality affects growth and regulation of the level of endogenous IAA of G. lucidum mycelium.
Fungal Polysaccharides ; analysis ; Indoleacetic Acids ; metabolism ; Light ; Peroxidases ; metabolism ; Reishi ; growth & development ; metabolism

OBJECTIVETo study the effect of light quality on growth, polysaccharides content of Ganoderma lucidum.

METHODG. lucidum was planted under different light qualities. The growth of G. lucidum was observed and polysaccharides content was determined in different growth periods.

RESULTThere were significant differences in the form of the thickness and the numbers of ringed of the fruit before the ejection. The maximum content of G. lucidum polysaccharides under different light qualities were appeared in different periods. The content of polysaccharides was higher than that of CK under blue light in the period of budding, parachute phase, after spore ejection. The content of polysaccharides was increased under green light in the period of growth stage. The yield of fruits and spores were different in different light qualities.

CONCLUSIONThe light quality could increase the content of polysaccharides in G. lucidum.

Light ; Plant Extracts ; analysis ; metabolism ; Polysaccharides ; analysis ; metabolism ; Reishi ; growth & development ; metabolism ; radiation effects

Lanosterol synthase( LS) is a key enzyme involving in the mevalonate pathway( MVA pathway) to produce lanosterol,which is a precursor of ganoderma triterpenoid. And the transcriptional regulation of LS gene directly affects the content of triterpenes in Ganoderma lucidum. In order to study the transcriptional regulation mechanism of LS gene,yeast one-hybrid technique was used to screen the transcription regulators which interact withthe promoter of LS. The bait vector was constructed by LS promoter,then the vector was transformed yeast cells to construct bait yeast strain. One-hybrid c DNA library was constructed via SMART technology. Then the c DNA and p GADT7-Rec vector were co-transformed into the bait yeast strain to screen the upstream regulatory factors of the promoter region of LS by homologous recombination. Total of 23 positive clones were screened. After sequencing,blast was performed against the whole-genome sequence of G. lucidum. As a result,8 regulatory factors were screened out including the transcription initiation TFIIB,the alpha/beta hydrolase super family,ALDH-SF superfamily,60 S ribosomal protein L21,ATP synthase β-subunit,microtubule associated protein Cript,prote asome subunit β-1,and transaldolase. Until now,the regulation effect of these 8 regulatory factors in G.lucidum has not been reported. This study provides candidate proteins for in-depth study on the expression regulation of LS.
Gene Library ; Intramolecular Transferases/metabolism* ; Reishi/genetics* ; Saccharomyces cerevisiae ; Transcription Factors/metabolism*

The synthetic characteristic of extracellular polysaccharide (EPS) of Ganoderma lucidum performed in batch fermentation was studied. The result showed that the production EPS was partially growth-associated. The cell dry weight (CDW) and EPS reached 15.56 g x L(-1), 3.02 g x L(-1) respectively. The yield of EPS to cell dry weight ( Y(p/x)) was 0.19. Based on the test result of batch fermentation, a kinetic model was proposed by using the Logistic equation for cell growth, the Luedeking Piret equation for EPS production and the Luedeking-piret-like equation for consumption of glucose as substrate. The calculated results of models were compared satisfactorily with experimental data under various glucose concentrations, the average relative error was no more than 5%. The kinetic model had practically guiding producing PES in fermentation of Ganoderma lucidum.
Culture Techniques ; methods ; Extracellular Space ; metabolism ; Fermentation ; Kinetics ; Models, Biological ; Polysaccharides ; biosynthesis ; Reishi ; chemistry ; growth & development

We studied the effects of several medicinal insects on biosynthesis of polysaccharides from Ganoderma lucidum in submerged culture. The results showed that the medicinal insect, Catharsius molossus at 5 g/L significantly promoted the biosynthesis of intracellular polysaccharides (IPS) and extracellular polysaccharides (EPS) of G. lucidum, and compared with control, IPS and EPS yields markedly enhanced from (1.93 +/- 0.09) g/L to (2.41 +/- 0.12) g/L and (520.3 +/- 20.2) mg/L to (608.9 +/- 20.2) mg/L, respectively (P < 0.05). Both IPS and EPS consisted of five kinds of components, and IPS-1 and EPS-1 were the major components of IPS and EPS, respectively. Further separation studies showed that IPS-1 was made up of three single compounds, while EPS-1 was made up of two single compounds. There were no new components in both IPS and EPS obtained from G lucidum in submerged culture by the addition of the insect, C. molossus, suggesting the biosynthetic pathways of the major components of IPS and EPS had not been changed.
Animals ; Cockroaches ; chemistry ; Culture Media ; chemistry ; Fermentation ; Materia Medica ; pharmacology ; Polysaccharides ; biosynthesis ; chemistry ; Reishi ; growth & development ; metabolism

Polysaccharide is among the main active components of Ganoderma lucidum for tumor prevention and treatment. Howe-ver, it remains unclear whether it has synergy with tumor immunotherapy. This study evaluated the effect of G. lucidum polysaccharides(GLP) on the infiltration of T lymphocytes into tumor and the underlying mechanism, in order to provide a reference for its application in tumor immunotherapy. GLP were prepared by water extraction and alcohol precipitation combined with Sevag method and then given(intraperitoneal injection) to the mice bearing B16-F10 cells at 25, 50 and 100 mg kg~(-1), respectively, to evaluate the effect on tumor growth. The infiltration of CD3~+ and CD8~+ T cells and the expression of intercellular cell adhesion molecule-1(ICAM-1) in tumor were detected by immunohistochemistry. EA.hy926 cells were treated with 50, 100 and 200 μg·mL~(-1) GLP, and the expression of ICAM-1 was determined by Western blot. The adhesion of EA.hy926 cells treated with GLP was measured with fluorescence-labeled Jurkat cells. To analyze the mechanism based on NF-κB pathway, this study determined the protein levels of nuclear factor kappa-B(NF-κB) p65, alpha inhibitor of NF-κB(IκBα), p-NF-κB p65 and p-IκBα by Western blot. The results showed that GLP can significantly inhibit the tumor growth in mice bearing B16-F10 cells, promote the infiltration of CD3~+ and CD8~+ T cells in tumor, and increase the expression of ICAM-1 in tumor. Meanwhile, GLP could also enhance the expression of ICAM-1 in EA.hy926 cells, thus strengthen the adhesion to Jurkat cells, induce phosphorylation and protein degradation of IκBα, and raise the expression and phosphorylation level of NF-κB p65. These results suggested that GLP could promote the expression of ICAM-1 through NF-κB pathway and further enhance the infiltration of T lymphocytes into tumor, thereby inhibiting tumor growth. This study lays a foundation for the further application of GLP in tumor immunotherapy.
Animals ; Endothelial Cells/metabolism* ; Intercellular Adhesion Molecule-1/genetics* ; Mice ; NF-kappa B/metabolism* ; Neoplasms ; Polysaccharides ; Reishi ; Signal Transduction ; T-Lymphocytes ; Tumor Necrosis Factor-alpha

OBJECTIVETo investigate the changes of xanthine oxidase (XOD), myeloperoxidase (MPO) and mitochondrial succinate dehydrogenase (SDH) in the testis and the protective effect of ganoderma lucidum spores on the testicular tissue of rats with non-insu- lin-dependent diabetes mellitus (NIDDM).

METHODSFifty male Wistar rats were divided randomly into a model, a ganoderma and a normal control group, the first two groups injected with 2% STZ (25 mg/kg) through the peritoneum, and the last one with half-and-half sodium citrate/citrate buffer solution. Two weeks after normal diet, glucose tolerance tests were performed and the rats with abnormal glucose tolerance in the model and ganoderma groups received high-fat and high-carbohydrate food, the latter given ganoderma lycium spores (250 mg/kg x d) in addition, both for 10 weeks and all rats fed alone. Glucose tolerance tests were repeated 1 day before the end of the experiment and the testes of the rats were harvested for the determination of XOD, MPO and SDH.

RESULTSSDH was significantly lower (P < 0.05) while XOD and MPO significantly higher in the model group than in the ganoderma and control groups (P < 0.05). The model rats exhibited abnormal convoluted seminiferous tubules, indistinct parietal layers, decreased or abolished gonepoiesis, luminal peripheral fibrous tissue (interstitial substance) accrementition, basal lamina thickening, and vessel wall fibrous tissue accrementition and sclerosis.

CONCLUSIONGanoderma lucidum spores can protect the testis of diabetic rats by reducing free radical-induced damage to the testicular tissue and enhancing the activity of SDH.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Peroxidase ; metabolism ; Rats ; Rats, Wistar ; Reishi ; Spores, Fungal ; Succinate Dehydrogenase ; metabolism ; Testis ; metabolism ; Xanthine Oxidase ; metabolism

OBJECTIVETo observe the effect of Jisheng injection (JSI) in protecting heart.

METHODSIsolated heart of rat was preserved in modified Euro-Collins solution (mEC) containing JSI for 20 hrs, and that preserved in simple mEC was taken as control. Then Langendorff isolated rat heart perfusion was conducted. Forty minutes after perfusion, the cardiac function, coronary flow, myocardial water content were determined, and lactate dehydrogenase (LDH), creatine kinase (CK) activity in perfusate, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content in myocardial tissue and pathologic change in myocardium were also observed.

RESULTSThe cardiac function and coronary flow of isolated heart preserved in JSI containing mEC was significantly better than those in the control (P < 0.01), with the LDH, CK activity and MDA content significantly lower (P < 0.01 and P < 0.05), SOD activity significantly higher (P < 0.05) and pathologic injury milder than those in control, but comparison of cardiac water content between the two groups showed insignificant difference.

CONCLUSIONJSI has good cardiac protective and anti-oxidizing effects.

Animals ; Antioxidants ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Heart ; Male ; Organ Preservation ; Organ Preservation Solutions ; pharmacology ; Rats ; Rats, Wistar ; Reishi ; chemistry ; Superoxide Dismutase ; metabolism

AIMTo study the antioxidant effect of Ganoderma polysaccharide peptide (GLPP) and its mechanism.

METHODSCopper was used as oxidant to induce low lipoprotein (LDL) oxidative modification, and alloxan was given i.v. to induce reactive oxygen species (ROS) injury in mice.

RESULTSGLPP decreased oxidation of LDL and the relative electrophoretic mobility (REM) of oxidative product of LDL. After GLPP was given i.p. for 20 days, the concentration of malondialdehyde(MDA) in serum and heart of mice was decreased. The GSHpx enzyme activity was increased, while the SOD level was decreased. The catalase(CAT) levels were not significantly changed by GLPP.

CONCLUSIONGLPP showed antioxidant effect by scavenging ROS or enhancing the enzyme activity of GSHpx in vivo and in vitro.

Animals ; Antioxidants ; isolation & purification ; pharmacology ; Glutathione Peroxidase ; metabolism ; Lipoproteins, LDL ; metabolism ; Male ; Malondialdehyde ; blood ; metabolism ; Mice ; Myocardium ; metabolism ; Oxidation-Reduction ; drug effects ; Plants, Medicinal ; chemistry ; Proteoglycans ; isolation & purification ; pharmacology ; Random Allocation ; Reishi ; chemistry ; Superoxide Dismutase ; metabolism