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15 March 2009, Volume 28 Issue 2
    

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  • Mycosystema. 2009, 28(2): 161-170.
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    Phylogenetic relationships of Dothideales and its allies were investigated based on analyses of nrDNA 18S (44 sequences of 31 species belonging to 20 genera) and 28S (41 sequences of 33 species belonging to 18 genera) partial sequences. The results indicate that the dothideomycetous genera tested are monophyletic and three lineages are recognized, i.e. Botryosphaeriales, Dothideales and Pleosporales; which supports the recognition of Botryosphaeriales as a separate order from Dothideales. The monotypic genus Macrovalsaria previously of uncertain taxonomic position turns out to be a member of Botryosphaeriales and seems to be related to Botryosphaeria. The placement of Neopeckia and of Dothidotthia is in Dothideales and Botryosphaeriales, respectively. The genera Plowrightia and Botryosphaeria as currently defined are not monophyletic. Species of Plowrightia appear to be associated with three genera, Dothidea, Neopeckia and Sydowia; those of Botryosphaeria tested are recognized as three groups. Although the seven genera of Pleosporales tested form a monophyletic group, bootstrap supports of all the phylogenetic trees obtained are very low; which suggests they may be recognized at a family or higher taxonomic rank. Sequence analyses support the previous placement of Phragmogibbera in Pleosporales based on morphological features.
  • Mycosystema. 2009, 28(2): 171-174.
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    Two loculoascomycete marine fungi were reported from the driftwood and sand wood collected from the coast of Shandong, China. Passeriniella obiones and Tremateia halophila were new Chinese records. Descriptions and illustrations of the species were given. The specimens were deposited in Mycology Herbarium at Qingdao Agricultural University (MHQAU).
  • Mycosystema. 2009, 28(2): 175-177.
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    During the study on foliicolous lichens collected from Hainan Province, a new species, Enterographa hainanensis B. Gao & J. C. Wei, is described in the present paper. A Latin diagnosis, Chinese description and discussion with 3 photographs are given.
  • Mycosystema. 2009, 28(2): 178-188.
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    Community structure and species diversity of the saxicolous lichens in forest ecosystem of mountainous area of southern Urumqi were analyzed using the principal component analysis (PCA) and cluster analysis methods. Forty-nine saxicolous lichen species belonging to 23 genera in 13 families of 4 orders were found in this ecosystem. According to the results of multivariate analysis and universal characteristics of habitat, the saxicolous lichen community in this forest ecosystem can be classified into three types: (1) Hygro-lichen community: Acarospora verruculosa + A. alphaplaca + Lecidea tessellata; (2) Xero-lichen community: Lecanora polytropa + Lecidea carpathica; (3) Alpino-lichen community: Verrucaria iaequalis + Dermatocarpon miniatum. The community 1 has highest species diversity (1.875) and community 3 has the lowest (1.147). Altitude and substrate types were important environmental factors influencing the structural patterns of saxicolous lichen community in this area.
  • Mycosystema. 2009, 28(2): 189-196.
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    A comparative study on biological characteristics was conducted to evaluate the difference of media, light duration, temperature and pH values on mycelial growth rate and morphological characteristics between two isolates S-C. f and G-C. f of Ceratocystis fimbriata causing pomegranate wilt and sweet potato black rot. The results indicate that the mycelial growth rate of the isolate S-C. f from pomegranate (Punica granatum) was much faster than the isolate G-C. f from sweet potato (Ipomoea batatas) in all experimental culture conditions. There was an obvious difference on colony morphology between the two isolates. The two isolates grew much faster on sweet potato dextrose agar (SPDA) as compared with those on other media and their growth rate was significantly higher in 24h darkness than that in 12h light and 12h dark alternation or 24h light. The isolates from pomegranate and sweep potato could survive in the temperature range of 10-36℃ and 10-33℃ respectively, and the optimum temperature was 25℃ for both of them. The pH range of mycelial growth was 4 to 12 and the most suitable pH value was 6. The mycelial lethal temperature was 50℃ in 10 minutes or 52℃ in 5 minutes for the isolate from pomegranate and 48℃ in 10 minutes or 50℃ in 5 minutes for that from sweet potato. Complete yeast extract medium could stimulate their sporulation. The mycelial growth rate was sharply decreased with the increasing glucose concentration in PDA medium.
  • Mycosystema. 2009, 28(2): 197-204.
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    The effects of metalaxyl, an acylalanine phenylamide fungicide exhibiting systemic properties, on morphology and ultrastructure of the wild-type isolate and the metalaxyl-resistant mutant of Phytophthora sojae in vitro were examined by means of electron microscopy techniques. Ultrastructral observations on the wild-type isolate revealed marked alterations at a low concentration of metalaxyl (1μg/mL), such as uneven growth of hyphae, excessive branching of hyphal tips, cessation of hyphal growth and bulb-like swelling of hyphal tips; mitochondria lost their distinct structures, number and size of vesicles increased, cell walls became thickened and hyphal cytoplasm was seriously degenerated in presence of a high concentration of metalaxyl (10μg/mL). While, hyphae of the metalaxyl-resistant mutant displayed little morphological changes after treatment with the high concentration of metalaxyl (10μg/mL). Observations indicated hyphal tips just appeared little excessive branching, hyphal cell contained a high number of lipid bodies, but fungal cell walls did not thicken. Hyphae maintained their growth and get across the filter treated with 10μg/mL metalaxyl.
  • Mycosystema. 2009, 28(2): 205-212.
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    The vegetative compatibility groups (VCGs) of Fusarium moniliforme were tested by applying nitrate nonutilizing (nit) mutant complementation technique to explore the population structure of the important pathogenic fungus. The 107 strains of F. moniliforme were isolated from cotton, corn and rice diseased tissues collected from Anhui, Shandong, Jiangsu and Hubei in China. The tested single-conidium strains were then transplanted on KClO3-containing potato sucrose agar (KPS) plates to mutagenize nit mutants and 1081 nitrate-nonutilizing (nit) mutants were recovered from the isolates by transferring chlorate-resistant sectors from KPS plates to minimal medium (MM). The four mutants types were identified as nit A, nit B, nit C and nit D based on the growing situation of the mutants on MM, nitrite medium (NM) and hypoxanthine medium (HM), three kinds of different nitrogen media. Among the four types, nit A was the most common one, with the frequency of 76%. Next were nit B and nit C, with the frequency of 12% and 10% respectively. The least one was nit D, 2%. The nit mutants then were paired on MM plates to test the vegetative compatibility of all isolates, according to their geographic location and hosts, respectively. Fifty-six different VCGs were identified totally from all three host-origin populations of the mutants. One sample was taken randomly from each VCG to determine the identity. The result showed isolates Fm1, Fm2 from cotton and isolates Fm19, Fm20 from corn belonged to the same VCG respectively, while the other strains did not. Similarly, 55 different VCGs were identified totally from all four locality-origin populations of the mutants. Only isolate Fm45 from Shandong and isolate Fm16 from Anhui belonged to the same VCG, while the other strains from different geographic origins belonged to different VCGs. The 107 tested isolates were divided into 54 VCGs, suggesting there was abundant diversity of VCGs in the population of Fusarium moniliforme isolates. The diversity analysis indicated that on the average, 1.9454 isolates from locality-origin population made up 1 VCG; the P and Shannon-Wiener diversity indices were 0.5140 and 1.0365, respectively; and 1.9107 isolates from host-origin population composed 1 VCG on an average; the P and Shannon-Wiener diversity indices were 0.5234 and 0.9048, respectively. The Shannon-Wiener diversity indices (H values) of the two populations were not significantly different from each other based on t test (t = 0.70, below t0.05 = 1.98). It was suggested that the VCG diversity of the two populations was not significantly different from each other, and the genetic similarity among the isolates from the same locality-origin was equal to that from the same host-origin.
  • Mycosystema. 2009, 28(2): 213-219.
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    Volvariella volvacea V23 and cold resistant mutant VH3 were treated at 4℃ for 0, 2, 4, 6, 8, 10h as templates, cDNA were synthesized by reverse transcription reaction with total RNA extracted from the mycelia, and plasmid contained Cor1 and GPD gene fragment was constructed as standard for real-time quantitative PCR to investigate the expression of Cor1 in strain V23 and VH3 treated under cold stress. The results showed that the expression of Cor1 at 4℃ reached the maximum at 0h, decreased gradually and reached the minimum at 6h, then increased gradually from 8h in strain V23; expression of Cor1 increased obviously and reached the maximum at 2h, then decreased at 4h, and increased gradually at 6h and 8h, but decreased again at 10h in VH3 during low temperature treatment at 4℃. The results indicated that Cor1 gene expression was correlated with ability for cold resistance in V. volvacea. The results will be helpful to further study on breeding new variety and genetic engineering in V. volvacea.
  • Mycosystema. 2009, 28(2): 220-225.
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    Cordycepin is an important secondary metabolite of Cordyceps militaris, a traditional Chinese medicinal fungus, and is mainly obtained from fruiting bodies at present. This article presents an efficient method of extracting and purifying cordycepin from rice medium, the waste of fruiting body production. The method included 3 steps: continuous countercurrent extraction, column chromatography with 732 cation exchange resin, and crystallization, and their working parameters were optimized. By use of the optimized method, cordycepin of purity over 98.0% was obtained, with the yield over 66.0%.
  • Mycosystema. 2009, 28(2): 226-235.
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    Four excellent isolates of Wolfiporia cocos with strong growth vitality, highly yielding ability, hereditary stability, good marketability and suitability for artificial cultivation in Guizhou were screened among 16 isolates from different sources. A screening model for excellent Wolfiporia cocos isolates was established.
  • Mycosystema. 2009, 28(2): 236-243.
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    Treated in water bath under 5 temperature gradient from 43℃ to 47℃ for 1min to 5min, the viability of Fusarium virguliforme, an exotic quarantine pathogen originated from USA and Argentina, was analyzed using Guava Easycyte Mini after fluorescence staining. The traditional spore germination test was set up as the control. The results showed that the pathogen was unvital treated with 47℃ for 4min. Statistical analysis using SPSS software indicated that very significant negative correlation was found between the pathogen viability and temperature. Similar result was also obtained between the viability and duration of treatment, except for strain 2-1 and 22825 showing significant negative correlation between viability and duration of treatment. Between spore germination rate and temperature as well as duration of treatment assumed very significant negative correlation, whereas between the fungus viability and the spore germination rate had very significant positive correlation. This study revealed that the traditional spore germination test could be replaced by the pathogen viability detection procedure, and the detection duration was greatly shortened and the detection efficiency was improved obviously.
  • Mycosystema. 2009, 28(2): 244-252.
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    The lignin oxidative system of 27 fungal species were analyzed qualitatively by four methods including using syringaldazine as substrate and aniline blue decolorization in plate cultivation. The result showed that 5 species had laccase (Lac), ligin peroxidase (Lip) and manganese peroxidase (Mnp), and 5 species simultaneously had Lac and Lip. Other 8 species only had one enzyme activity. Twelve species produced Lac. Lac activity of Fomes fomentarius reached a peak of 9496U/mL in inducted and static cultivation, which was obviously higher than that of other fungi. By static cultivation, the Lac activity of Hymenochaete badio-ferruginea and Irpex lacteus reached a peak of 652 and 292U/mL, respectively, which were higher as compared with other cultivation methods. It proved that static cultivation could take the place of shaking cultivation in the production and preparation of Lac of Fomes fomentarius, Hymenochaete badio-ferruginea and Irpex lacteus. In the decolorization assay, it was found that Fomes fomentarius, Hymenochaete badio-ferruginea, Irpex lacteus, Phellinus igniarius and Trichaptum fusco-violaceum had efficient and broad-spectrum discolored function to congo red, alizarin red, methylene blue and crystal violet.
  • Mycosystema. 2009, 28(2): 253-260.
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    The characteristics of culture, xylanase producing condition and characteristics of enzyme of Geotrichum candidum Ref1 were studied. The result showed that Ref1 was a psychrophile, and the best growing conditions were pH 6, 20℃ and yeast extract as nitrogen source while the best xylanase producing conditions were pH 3-7, 15℃ and yeast extract as nitrogen source. Xylanase activity of optimized Ref1 reached 118.7U/mL and soluble protein 60?g/mL; the specific activity of the crude enzyme was 1250U/mg protein. The xylanase has an optimal activity at 50℃ and pH 5.0, and the relative enzyme activity remains 80% while keeping at the temperatures of 15-40℃. Mg2+, Na+ and 8mmol/L Fe2+, Cu2+, Zn2+ were strong inhibitors, while Ca2+ , 4mmol/L Fe2+, Cu2+, Zn2+ and 8mmol/L Mn2+ were stimulators of the enzyme. The maximum velocity of reaction and the Km of the xylanase were 163.38mmol/mg/min and 0.75mg/mL, respectively.
  • Mycosystema. 2009, 28(2): 261-267.
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    The reaction conditions and primers of Armillaria gallica phylogeny analysis based on ISSR-PCR were studied. The annealing point of every primer and the influence of formamide were analyzed especially. The result provided a consult for studying on phylogenesis, genetic diversity and germplasm resource of Armillaria biological species.
  • Mycosystema. 2009, 28(2): 268-274.
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    Thermomyces lanuginosus produces an extracellular lipase that is important for industrial applications. Based on published DNA sequence, we cloned the full length genomic DNA and cDNA of the lipase using PCR and RT-PCR methods. DNA sequencing revealed that the genomic DNA of the T. lanuginosus lipase gene has an open reading frame of 876bp encoding a protein lipase precursor of 292 amino acid residues, and three introns. And the cDNA of lipase gene we cloned is identical to the registered lipase gene of Thermomyces lanuginosus in GenBank (AF054513). The DNA and cDNA sequence have been registered in GenBank with accession number EU022703 and EU370914, respectively. PCR product consisting of the cDNA of the T. lanuginosus lipase gene coding region without its signal sequences, first 17 amino acid residues, was cloned into the yeast secretive plasmid pPIC9K, which was then transformed into Pichia pastoris GS115 strain. The T. lanuginosus lipase gene was expressed and recombinant lipase was secreted into the culture medium in an active form. Production of recombinant protein reached 0.93g/L when it was induced by 1% (V/V) methanol at 30℃ in small-scale cultures. The recombinant enzyme exhibited optimum activity 7.2U/mL at pH 8.0 and 60℃. It is a thermostable enzyme since it retained complete activity in 1h at 60℃. The effective expression of the recombinant lipase in P. pastoris makes it become an attractive system to produce enzyme for industrial applications.
  • Mycosystema. 2009, 28(2): 275-282.
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    The SSR genotypes of 66 isolates of Phytophthora infestans (potato late blight pathogen) collected from potato in China and 2 reference isolates were identified and nominated by use of two loci, Pi4B and Pi4G. Among 66 isolates of P. infestans, seven SSR genotypes (D-03, D-05, D-06, G-02, H-01, F-01 and F-06) were found, of which F-06 was a novel one. Fifty-three isolates, accounting for 80.3% of the total were F-01, which was the predominant in China. Statistic analysis of allele genes generated by two loci (Pi4B and Pi4G) indicated that diversity produced by the locus Pi4B were higher than that by Pi4G. SSR data showed that genetic diversity of isolates from Hebei province was mostly the same as that of isolates from Heilongjiang province. However, genetic diversity of isolates from Yunnan province was significantly different from that of Hebei and Heilongjiang. In addition, the results showed that SSR genotype of P. infestans did not correlate to phenotype of metalaxyl-resistance.
  • Mycosystema. 2009, 28(2): 283-288.
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    The full-length cDNA of Bbhsp70 gene was cloned from Beauveria bassiana using SMART RACE RT-PCR, and its upstream sequence was amplified using genome walking. Analysis of the cloned complete cDNA with a whole length of 2405bp showed that it encompassed a open reading frame (ORF) with 1971bp encoding 656 amino acid. The mature protein had a molecular mass of 71.3kDa with a calculated pI of 4.92. The analyses indicate that the upstream sequence contained several regulatory elements, such as CCAAT-binding factor, GC-box, HSE and GATA, but the TATA-box and CAAT-box could not be found. The findings of this study are essential for further functional investigation of hsp70 gene of Beauveria bassiana and construction of engineered strain with enhanced anti-adversity ability.
  • Mycosystema. 2009, 28(2): 289-294.
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    Differential-display reverse transcription-PCR (DDRT-PCR) was used to investigate the difference of gene expression between unfruitificative mutant and wild type strain of Lentinula edodes, and 21 cDNA fragments were obtained. Twelve fragments showed high sequence similarity to known gene products, including ubiquitin, ATP synthase, zinc finger family protein, GTP binding protein, elongation factor g1 mitochondrial precursor, peroxiredoxin, thioesterase family protein, 2,4-dienoyl-CoA reductase, pyridoxal-phosphate dependent TrpB-like enzyme, glycoside hydrolase family 65, heat shock protein and hydrophobin. The genes indicate that the fruiting body development in L. edodes is a complex process involving several pathways and signal transduction such as intracellular trafficking, transcriptional regulation, cell division, protein synthesis, metabolism, and stress response.
  • Mycosystema. 2009, 28(2): 295-298.
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    This paper deals with different immunological regulation effects of Phellinus igniarius crude polysaccharides on tumor-bearing mice. The tumor bearing mice were divided into four groups at random: crude exo-polysaccharide treatment group, mycelium crude polysaccharide treatment group, fruit body crude polysaccharide treatment group and negative group. The treated mice were killed in 10 days and the activity of natural killer cells (NK) and the lymphocyte proliferation were investigated. The results indicate that as compared with negative group, all of the tested groups could enhance the activity of NK cells and the lymphocyte proliferation (P<0.01), suggesting that both the fermented and the fruit body crude polysaccharides of Phellinus igniarius had regulatory effects on the immunity of tumor bearing mice.
  • Papers
  • Mycosystema. 2009, 28(2): 299-302.
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  • short communications
  • Mycosystema. 2009, 28(2): 303-309.
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  • Mycosystema. 2009, 28(2): 310-314.
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