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15 November 2009, Volume 28 Issue 6
    

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  • Mycosystema. 2009, 28(6): 765-768.
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    Ajuga multiflora (Labiatae) is an ornamental plant widely growing in gardens in southern China. Blackleg symptoms were observed on A. multiflora in autumn of 2005 in Hangzhou, Zhejiang Province. It was caused by Phoma species. The disease was found throughout the year, and most severe during the flowering phase. Brown water-soaked spots initially appeared on the base of leaf petioles, the lesions rapidly became black-rot, then followed by wilting and drying of individual leaves. At last the disease resulted in plant death. The typical symptom of the disease was black stem rot. The six isolates of the causal agent were all identified as Phoma multirostrata based on their morphological characteristics, pathogenicity tests and rDNA ITS sequence analyses. It is the first report of P. multirostrata causing blackleg of A. multiflora in China.
  • Mycosystema. 2009, 28(6): 769-775.
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    In order to obtain the hexanol-degrading fungi, the methods of culture enrichment and UV induction were used in this study. To isolate useful fungi, different materials including apple orchard soil, apple pomace, apple cider mash and vinegar dregs were tested and two valuable fungal strains of TF and TM were obtained from apple orchard soil. The strains showed high ability in degrading hexanol at pH3.8-4.0. During growth in potato dextrose broth (PDB) containing 4.0mg/mL hexanol for 7 days, TM and TF degraded hexanol by 45.60±5.43% and 23.82±9.27%, respectively, significantly different from the control. Based on the morphological characteristics and sequence analysis of 26S rDNA D1/D2 for strain TF and nrDNA ITS for strain TM, TM was identified as Geotrichum sp. and TF as Geotrichum candidum (telemorph, Galactomyces geotrichum).
  • Mycosystema. 2009, 28(6): 776-782.
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    Brome grasses are economically and ecologically important forage grasses, and have been frequently introduced into China these years. The smut fungi Tilletia spp. are important pathogens on these grasses and have greatly endangered grass production. There are four Tilletia species, i.e. T. controversa (TCK), T. bromi, T. bolayi and T. caries (TCT), infecting these grasses. It is difficult to separate them from each other based on the characteristics of their teliospores. In this study, the morphology, autofluorescence and germination behavior of teliospores were determined and four species, T. bromi, TCK, TCT and T. fusca with similar characteristics were selected for further analysis. Based on the nucleotide difference of a beta-tubulin tub2 gene, a molecular marker that is sequence characterized amplified region (SCAR) was developed specifically for PCR detection of T. bromi by using the DNA samples from either the mycelia or the teliospores isolated from brome grasses. The method developed in this study can be quickly and reliably employed in quarantine analysis.
  • Mycosystema. 2009, 28(6): 783-789.
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    Collema tenax and Peltigera didactyla are N-fixing lichens in arid and semiarid grasslands, where water and nitrogen availability are essential factors influencing ecosystem structure and functions. The response of N-fixing activity of lichens to drought in temperate grasslands of China has received little attention in the past ecological studies. The purpose of the present study was to investigate the effects of drought duration on N fixation of C. tenax and P. didactyla by measuring potential N-fixing activity of both lichens that have been subjected to drought for 0, 1, 2, 4, 8 and 16 days. All samples were collected from a semiarid grassland in Inner Mongolia and N-fixing activity was measured using acetylene reduction assay. Most drought duration treatments yielded insignificant differences in N-fixing activity between species. N-fixing activity of both C. tenax and P. didactyla sigmoidally decreased with drought duration (R2 = 0.9996 and R2 = 0.9954, respectively). These results show an inhibitory effect of drought on N fixation for both lichens. This effect can be intensified by prolonged drought duration. No significant changes in N fixation of both lichens were found in the drought duration less than 2 days. However, a 4-day drought resulted in a significant decline in N-fixing activity of C. tenax and P. didactyla (decreased by 34% and 54%, respectively). N-fixing activity of both lichens decreased by up to 90% when being dried for 8 days, and declined by more than 99% in 16-day drought duration. The negative effects of drought on N fixation of both lichens could be attributable to the low levels of photosynthate and energy, and to the lag time after rewetting before N-fixation was initiated and before maximal N-fixing activity was reached.
  • Mycosystema. 2009, 28(6): 790-796.
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    In order to isolate the drought-resistant gene from the mycobiont of desert lichens, the desiccation-tolerance biological research and liquid culture were performed. The mycobiont isolated from the desert lichen Glypholecia scabra, was studied under the water stress. The results showed that the mycobiont isolated from G. scabra would accumulate soluble sugars and increase the concentration of total phenols and reduce glutathione (GSH) to protect itself from low water activities. It could be inferred that the genes relating to the metabolic pathways of phenols and GSH, may likely to be the candidates of drought-tolerance genes for cloning and expression analysis at aw=0.90 and aw=0.85. Additionally, in order to provide enough materials for cloning and expression analysis, optimal liquid culture conditions of the mycobiont isolated from G. scabra were studied using Plackett-Burman design and orthogonal arrays. The optimized medium (pH6.3) consisted of 15g glucose, 2g soy peptone, and 200μg Vitamin B12 in 1L potato extract. 150mL medium filled in 250mL triangular flask was shaken under revolving speed of 100r/min, at 18℃. The growth curve showed that the biomass of the mycobiont isolated from G. scabra was improved significantly under the optimized conditions.
  • Mycosystema. 2009, 28(6): 797-801.
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    A binary expression vector of thermotolerance-related gene 028-1 of Agaricus bisporus was constructed and transferred into non-thermotolerance strain 8213 of A. bisporus by Agrobacterium-mediated transformation. After hygromycin resistance selection and PCR identification, dozens of transgenic strains of A. bisporus were obtained. 10 of them were put into compost running test under different temperatures, and the results showed that the thermotolerance of most of the transgenic strains increase obviously.
  • Mycosystema. 2009, 28(6): 802-805.
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    The total protein extraction effect of Ganoderma lucidum primordium by Tris-saturated phenol law and trichloroacetic acid (TCA)/acetone precipitation methods were compared. After analyzing the two-dimensional electrophoresis (2-DE) protein atlas by the software of Image Master 2D Platinum 6.0, 565 and 273 protein spots were obtained separately by the method of Tris-saturated phenol and TCA/acetone precipitation; The 2-DE gels of TCA/acetone precipitation showed strong horizontal streaks in the atlas of alkalinity end and low molecular region, while the Tris-saturated phenol law was capable of generating more protein spots, obtaining better effect of isoelectric focusing and reducing salt. Tris-saturated phenol law can be considered as a feasible protein extraction method for Ganoderma lucidum primordium and the results of this study are of great significance for developing effective protein extraction methods in other medicinal mushrooms and will lay the foundation of further study on the discrepancy expression of Ganoderma lucidum treated by bi-directional solid fermentation.
  • Mycosystema. 2009, 28(6): 806-812.
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    The expression of hsp70 gene from Beauveria bassiana under several stress conditions was detected using Realtime-PCR, and the effects of different stress conditions on expression of hsp70 gene on mRNA level were explored. The results showed that under high temperature stress (38℃), the expression level of hsp70 gene reached the maximum (10.18 times of the control) in 30 minutes, and then decreased gradually. It reached the minimum (2.85 times of the control) in 180 minutes. Under low temperature stress (4℃), the expression level of hsp70 gene reached the minimum (0.25 times of the control) in 2 hours, and then increased gradually. It keep stable (1.4-1.5 times of the control) in 10 hours. Under UV stress (253.7nm wavelength), the expression level of hsp70 gene reached the maximum (2.33 times of the control) in 3 minutes, and then decreased rapidly. It keep stable (0.2 times of the control) in 60 minutes. It was inferred that hsp70 gene in Beauveria bassiana might play an important role in this three stress tolerances. The results also showed that hsp70 gene promoter of Beauveria bassiana could make gene high express in stress conditions. Therefore, it may have great application value in construction of engineered strain with enhanced anti-adversity ability.
  • Mycosystema. 2009, 28(6): 813-818.
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    The water-soluble crude polysaccharide (APP) was isolated from fruit bodies of Auricularia polytricha by hot water extraction, ethanol precipitation, and deproteination with Sevage method. It was found that APP could markedly inhibit growth of Sarcoma 180 in mice. APPⅠ, Ⅱ and Ⅲ was separated from APP by ion-exchange on DEAE-Cellulose-52, and APPⅡ could significantly exhibit the anti-tumor effect. APPⅡ was further purified by gel-filtration chromatography on Sephacryl S-300 columns and separated into APPⅡA and APPⅡB. It was confirmed that APPⅡA could significantly inhibit tumor growth. APPⅡA was characterized by thin-layer chromatography (TLC), agarose gel electrophoresis (AGE), high performance liquid chromatography (HPLC), and UV, IR and 1H-NMR spectroscopy. The results demonstrated that APPⅡA was homogeneous and its relative molecular weight was approximately 110,000Da. Monosaccharide analysis showed that APPⅡA was a heteropolysaccharide which was composed of xylose and mannose. The glycosidic linkages of APPⅡA was estimated to contains both α- and β- pyranoside form.
  • Mycosystema. 2009, 28(6): 819-824.
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    The effects of Phaeoporus obliquus polysaccharides on apoptosis of hepato-carcinoma HepG-2 cells were investigated. The inhibition of cell growth was determined by mono-nuclear cell direct cytotoxicity assay (MTT); morphological changes were observed by transmission electron microscopy; apoptosis was assessed by DNA ladder and flow cytometry (FCM). The mRNA expression levels of Bcl-2 and Bax genes were detected by reverse transcription-polymerase chain reaction (RT-PCR). Phaeoporus obliquus polysaccharides inhibited the growth of HepG-2 cells in vitro in dose-and time-dependent manner. Microscopic observation showed that the cells presented some morphologic features of apoptosis. Agarose gel electrophoresis analysis showed DNA ladder; hypodiploid peak appeared on flow cytometry (FCM) histogram, and the apoptotic rate of HepG-2 cells was risen with the increase of dose. The Bax mRNA expression was remarkably up-regulated in HepG-2 cells exposed to Phaeoporus obliquus polysaccharides, but no apparent changes of Bcl-2 gene were observed. Phaeoporus obliquus polysaccharides inhibit proliferation of HepG-2 cells and induce apoptosis. This may associate with upregulating of the expression of Bax gene.
  • Mycosystema. 2009, 28(6): 825-831.
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    To obtain a low-cost medium for submerged fermentation of Ganoderma lucidum in a large scale, four medium constituents were optimized by using central composite rotatable design (CCRD) and response surface methodology (RSM). The four factors were corn flour (x1), wheat bran flour (x2), soybean flour (x3) and sucrose (x4). The results show that the mathematical model for biomass production and polysaccharide production are Y1 = 15.1 – 0.31x1 – 0.34x2 + 0.36x3 – 0.44x4 – 1.26x12 – 1.98x22 – 0.85x32 – 1.15x42 – 0.59x2x3, and Y2 = 2.0 – 0.08x1 – 0.08x2 + 0.04x3 – 0.09x4 – 1.13x12 – 0.33x22 – 0.08x32 – 0.16x42 – 0.16x2x3 – 0.10x1x4, respectively. Based on the models, the optimum medium for biomass production is corn flour 19.7g/L, wheat bran flour 11.3g/L, soybean flour 6.3g/L, and sucrose 19.5g/L; while the optimum medium for polysaccharide production is corn flour 19.6g/L, wheat bran flour 11.0g/L, soybean flour 6.7g/L, and sucrose 19.1g/L. The maximum biomass and polysaccharide in a 150-L stirred-tank bioreactor reach 16.92g/L and 1.86g/L under the optimized medium, respectively. The use of the medium can reduce the costs for both biomass and polysaccharide production by G. lucidum at large scale.
  • Mycosystema. 2009, 28(6): 832-837.
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    The effects of acidic extracts from Ganoderma lucidum fermentation broth on chronic bronchitis in mice were investigated by expectorant, anti-tussive experiments and morphological observations. Ganoderma lucidum fermentation broth (GFB), different dosages of ethanol-extracts and butanol-extracts of GFB significantly prolonged the latent period of cough induced by SO2 and reduced the frequency of coughing, exhibiting obvious expectorant effect. Butanol-extracts with a dosage of 60mg/kg improved the excretion of phenol red by 50.7% as compared with controls, showing distinct antitussive effect over Honeysuckle-Scutellaria-Forsythia extract mixture oral liquid oral liquid. Ganoderic acidic extracts obviously alleviated the bronchial trauma of smoked mice, and the pathomorphological change was noticeably decreased as compared with the model group. However, the anti-inflammatory and anti-injury effects were equivalent to Honeysuckle-Scutellaria-Forsythia extract mixture oral liquid, which may provide the pharmacological basis for the development of Ganoderic acidic products.
  • Mycosystema. 2009, 28(6): 838-845.
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    The in vitro anti-tumor activity of acidic triterpenoids from Ganoderma sinense mycelia and submerged fermentation liquid by using 3-(4, 5-dimethylthiozol -2-yl)-2, 5-diphenyltetrazolium bromide (MTT) method, as well as the antimicrobial activity on several bacteria and molds by using cylinder-plate method were investigated. The results showed that both the intracellular and extracellular acidic triterpenoids at 250μg/mL displayed significant inhibitory effects on human hepatoma cell line BEL7402 and human breast carcinoma cell line MCF-7 (P < 0.05); however, they had no significant inhibitory effects on human gastric cancer cell Line SGC-7901. The growth of BEL7402 cells treated by intracellular acidic triterpenoids was significantly inhibited, and there was no exponential phase in the growth curve. After 3d culture, BEL7402 cells treated by intracellular acidic triterpenoids appeared fewer and smaller as compared with the cells in control. The acidic triterpenoids at 40mg/mL had significantly inhibitory effect on the growth of Escherichia coli and Staphylococcus aureus (P < 0.01), however, the sample had little inhibitory effect on Bacillus subtitis and Penicillium chrysogenum, and had no inhibitory effect on Aspergillus niger. Further studies revealed that the minimal inhibitory concentration (MIC) of the acidic triterpenoids to E. coli, S. aureus, B. subtitis, A. niger and P. chrysogenum was 20mg/mL, 20mg/mL, 40mg/mL, 80mg/mL and 40mg/mL, respectively. The antibacterial components of the acidic triterpenoids sample couldn’t be destroyed in 60℃ treatment for 2h, but it could be partly destroyed in 80℃ treatment for 2h, thus the antimicrobial activity was decreased.
  • Mycosystema. 2009, 28(6): 846-849.
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    This paper deals with antioxidant ability of methanol extract from 4 Antarctic lichens, i.e. Usnea aurantiacoatra, Usnea antarctica, Sphaerophorus globosus and Cladonia sp. The antioxidant ability was detected by the linoleic acid, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and reducing power systems, and compared with that of commercial antioxidant Vitamin C and butylated hydroxyanisole (BHA). The results showed that the lichens collected from Antarctica exhibited high antioxidant activities, and the antioxidant activity was concentration-dependent. Among 4 lichens, Usnea aurantiacoatra had the maximum inhibitory ratio of lipid peroxidantion and reducing power, Cladonia sp. showed the maximum scavenging activity on DPPH free radical. Besides, usnic acid from Usnea aurantiacoatra also exhibited antioxidant activity, however, its activity was lower than that of the crude extract from Usnea aurantiacoatra.
  • Mycosystema. 2009, 28(6): 850-856.
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    真菌,预处理,生物吸附,解吸附,吸附机理,电镀废水
  • Papers
  • Mycosystema. 2009, 28(6): 857-859.
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  • Mycosystema. 2009, 28(6): 860-862.
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  • Mycosystema. 2009, 28(6): 863-864.
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  • Mycosystema. 2009, 28(6): 865-869.
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  • Mycosystema. 2009, 28(6): 870-872.
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  • Mycosystema. 2009, 28(6): 873-877.
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  • short communications
  • Mycosystema. 2009, 28(6): 878-887.
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  • Mycosystema. 2009, 28(6): 888-894.
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  • Mycosystema. 2009, 28(6): 895-901.
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