15 July 2009, Volume 28 Issue 4

    

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  A new species of the genus Lophodermium on Pinus armandii

  Mycosystema. 2009, 28(4): 473-475.

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  A new species of Lophodermium, Lophodermium erlangshanense sp. nov. on needles of Pinus armandii from Erlangshan in Sichuan Province, is reported. Latin and Chinese descriptions and illustrations are provided. The type specimen is deposited in the Forest Mycology Herbarium of Sichuan Agricultural University (FMSAU).
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  Notes on dematiaceous hyphomycetes from soil in Shanxi Province, China Ⅰ

  Mycosystema. 2009, 28(4): 476-482.

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  A total of 49 isolates of soil dematiaceous hyphomycetes belonging to 36 species in 27 genera were found from Shanxi Province. Among them, Alternaria pratensis is a new species, while Chaetopsina is a newly recorded genus for China; Chaetopsina fulva, Cordana oblongispora, Humicola brevis, Leptographium lundbergii and Memnoiella subsimplex are newly recorded species for China. The other 30 previously reporded species from China are also listed. Specimens (dried cultures) and living cultures of all fungi studied have been deposited in the Herbarium of Shandong Agricultural University: Plant Pathology (HSAUP).
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  Wood-inhabiting marine fungi from the coast of Shandong, China Ⅶ

  Mycosystema. 2009, 28(4): 483-489.

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  Four species in Halosphaeriales were observed on the incubated drift and intertidal woods collected from the coasts of Yellow Sea and Bohai Sea. Ascospores of these species are ellipsoidal, bicelled and hyaline. Morphology of the four species was discussed in the paper. The specimens and cultures were deposited in Mycology Herbarium at Qingdao Agricultural University (MHQAU).
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  Culture study of Marssonina coronaria from diseased apple leaves

  Mycosystema. 2009, 28(4): 490-495.

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  Marssonina coronaria is the causal agent of Marssonina leaf blotch of apple which leads severe defoliation starting at early summer. It is difficult to obtain the pure culture of this fungus because of its slow growth. Three isolation methods were compared by using 4 culture media in this study. The results showed that each isolation method was successful to obtain the pure culture of fungus. However, single spore isolation was much easier, less contamination and more successful than isolation using diseased leaf segment and conidia mass from acervuli. The isolation methods of the latter only showed successful rate of 10%. Additionally, the growth of the pathogen was obviously affected by cultural media. The morphological characters such as colony color and diameter, and production of fruit body of the fungus growing on PDA, 10% V8, apple leaves dextrose agar (LDA) and apple leaves extraction dextrose agar (LEDA) media were different. The fungus formed colonies of about 8mm in diameter and did not produce fruit bodies and aerial hyphae in 1 month incubation on PDA. However it formed the similar size colonies as on PDA and produced fruit bodies and aerial hyphae on 10% V8 and LDA media in the same incubation period. Very small colonies (2mm in diameter) and fruit bodies were found on LEDA media in the same incubation condition. These results indicated that successful isolation of M. coronaria from apple leaves depended on suitable isolation method and cultural media as well as fresh samples.
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  SSR analysis of population genetic diversity of Puccinia striifornis f. sp. tritici in different altitude regions of Longnan, Gansu

  Mycosystema. 2009, 28(4): 496-503.

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  Stripe rust (or yellow rust) of wheat (Triticum aestivum), caused by Puccinia striiformis f. sp. tritici, is one of the most important diseases of wheat worldwide. It is very important to research the population genetic diversity of P. striifornis f. sp. tritici in different altitude regions of Longnan, one of the largest and the most important over-summering areas in China. Using the TP-M13-SSR, population genetic diversity and variation of P. striiformis f. sp. tritici, containing 330 isolates 11 collections from different altitude area of Longnan, were investigated. The important conclusion is that P. striifornis f. sp. tritici populations from different altitude areas were genetically different. Genetic diversity for the pathogen population from high mountainous area is much higher than that from mountainside area or lowlands. The genetic differentiation for the populations obtained from high mountain and mountainside areas was higher than that of the population from the lowlands, suggesting the limited gene flow in mountain area, and extensive gene exchange in lowlands. Our molecular data confirm that the stripe rust pathogen is able to exchange between lowland areas and high mountain areas in the region.
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  Diversity of endophytic fungi in leaves of Ginkgo biloba

  Mycosystema. 2009, 28(4): 504-511.

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  Diversity of endophytic fungi in leaves of Ginkgo biloba was studied in order to understand diversity state and succession change of endophytic fungal communities in this endemic plant. Healthy leaf samples were collected from five main planting locations of G. biloba in Jiangsu and Shandong provinces separately in spring, summer and autumn. A total of 1971 strains of endophytic fungi was isolated. As a result of morphological identification 32 genera were obtained, of which Alternaria (amounting to 18.01% of the total), Phyllosticta (14.66%) and Rhizoctonia (14.56%) were dominant. The distribution and composition of endophytic fungi varied with plant organs colonized, localities and seasons, but they assumed regularity to a certain extent. Populations of endophytic fungi in different parts of the leaves indicated different abundance and uniformity but assumed similarity to a certain extent.
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  The community composition of the arbuscular mycorrhizal fungi in the rhizosphere of Phellodendron amurense

  Mycosystema. 2009, 28(4): 512-520.

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  The rhizospheric soil and root samples of Phellodendron amurense were collected in the Logging Station of Northeast Forestry University. Nested-PCR was conducted to specifically amplify NS31/Glol domain sequences of the small-subunit (18S) rDNA from the AM fungi in the root and rhizospheric soil samples of P. amurense, respectively. The Nested-PCR products were applied to denaturing gradient gel electrophoresis (DGGE) analysis. Then, the community composition of the AM fungi was analyzed by DGGE, sequencing, and phylogenetic analysis. The result indicated that the targeted product (230bp) was successfully amplified from trace DNA by Nested-PCR. There were differences in DGGE profiles such as band numbers, densities and dominance between the rhizospheric soil and root samples. All sequences were divided into three groups, viz. Glomus, Scutellospora and Hyponectria buxi. Glomus sp. (EF177624) and Glomus sp. (DQ085205) were the prevalent AM fungi in the root and rhizospheric soil, respectively.
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  The component of entomogenous fungi community and the niche of dominant species in pure forest of Masson’s pine

  Mycosystema. 2009, 28(4): 521-527.

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  The structure of entomogenous fungus communities of the artificial forest ecosystem of pure Masson’s pines in Magushan was investigated, and 3 families, 6 genera and 14 species were found. The main species of the community were Isaria cateinannulata, I. farinosa and Beauveria bassiana. The nutritional-niche, temporal-niche and nutritional-temporal two-dimension niche of these 3 dominant species were analyzed. The result showed that the largest nutritional niche overlap occurred between I. cateinannulata and I. farinosa, and the largest time niche overlap occurred between I. cateinannulata and B. bassiana. B. bassiana had the highest values in nutritional-niche breadth, temporal-niche breadth and two-dimensional niche width. The facts that B. bassiana has long occurring period, wide distribution, wide insect host ranges and strong survival ability in the forest support the results.
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  The ecological distribution and resources of the macrofungi in Huaping Nature Reserve of Guangxi, China

  Mycosystema. 2009, 28(4): 528-534.

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  Two hundred and one species of macrofungi were found from Huaping Nature Reserve of Guangxi, China, belonging to 105 genera in 45 families. Among them 50 species are edible, 38 medicinal, 13 poisonous, 59 mycorrhizal, and 48 wood-decaying. The macrofungi can be roughly classified into the following five ecological communities: 1) community in evergreen broadleaf forest; 2) community in deciduous broadleaf forest; 3) community in mixed deciduous evergreen broadleaf forest; 4) community in shrubbery; 5) community in bamboo forest. Three vertical distribution types are recognized, viz. lower-montane type, mid-montane type, and high-montane type.
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  Isolation, purification and structure of Cp-I toxin from Cryphonectria parasitica

  Mycosystema. 2009, 28(4): 535-540.

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  The strain of Cryphonectria parasitica was cultivated by liquid fermentation. The crude material with wilt-inducing activity to chestnut browse was obtained by using petroleum ether extracting method from the fermentation broth. Three compounds were obtained by means of silica gel column chromatography using mixed solution of V(chloroform ):V(petroleum ether ):V(alcohol) = 6:2:2 as eluent. The results showed the first compound (Cp-I) had higher wilt-inducing activity than the other two. The chemical formula of the pathogenic fraction Cp-I was identified as C16H22O4 (M=278) by mass spectrography (MS), nuclear magnetic resonance (NMR) and infrared spectrum (IR).
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  Genetic differentiation of self-crossing S1 generations of Pleurotus eryngii

  Mycosystema. 2009, 28(4): 541-547.

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  Strain 24 (Guang Xing) of Pleurotus eryngii was selected as self-crossed materials and comprehensive analysis of qualitative (color, deformity and growth characteristics) and quantitative (shape, yield, size and number of fruit bodies) traits of S1 generations was made. The results showed that 70% of inbred progenies fruited successfully. The average mycelium growth rate and yield of inbred progenies are lower than those of the parents’ strains. Average yield of 16% of generational strains are higher than that of parents and the highest average yields reach 204.0g which are higher by 45.7% than those of parents. The average yield was found to be indistinctly correlative (R = -0.028) with mycelium growth rate in sawdust medium, but strongly relative to fruit-body number (R = 0.543). Among the S1 generations, most pilei (52%) are concolorous with those of parent (gray). About 20% of strains are of non-parental shape (claviform) and deformative fruit-bodies (19%) were observed. The variation degree of pileus diameter is similar to that of stipe diameter and stipe length. No significant differences were observed among them. These characteristics can be used to select excellent strain in terms of breeding purposes.
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  Development of microsatellite markers for Lentinula edodes, using database search and ISSR-suppression PCR methods

  Mycosystema. 2009, 28(4): 548-552.

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  Two methods, database search and ISSR-suppression PCR, were used to isolate microsatellite markers from Lentinula edodes. Twenty-one primer pairs were designed by database search, and eleven of them showed polymorphism. The number of alleles produced by these eleven markers was 3.3 per locus in average. Using method of ISSR-suppression PCR, eight primer pairs were developed and five of them had polymorphism. There were 3 alleles per locus in average produced by these five markers. The results showed that two methods were effective and applicable to develop SSR from L. edodes.
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  Sequence and phylogenetic analysis of OPA2-1 region from Alternaria small-spored species

  Mycosystema. 2009, 28(4): 553-557.

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  The OPA2-1 region of small-spored Alternaria species was amplified with specific primer pairs, OPA2-1L/OPA2-1R, all strains produced one single same size band. The analysis of their nucleotide acid sequences indicates that certain differences were found in nucleotide sequences among these species. The phylogenetic tree constructed based on neighbor-joining method shows that these isolates were divided into eight clades. Alternaria gaisen, A. longipes and A. arborescens were clustered into different clades, while isolates of A. alternata were distributed into different groups. It shows that A. alternata should be treated as a species complex. It is found that clear variations were present in the OPA2-1 sequences of all tested small-spored Alternaria species, it provides a meaningful information for phylogenetics of small-spored Alternaria species.
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  Bioactivities of intracellular polysaccharide from Pleurotus citrinopileatus

  Mycosystema. 2009, 28(4): 558-563.

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  Extraction of the intracellular polysaccharide from Pleurotus citrinopileatus with hot water and antimicrobial activity and scavenging capacity to superoxide anion radical and nitroso of the polysaccharide were studied. The optimum conditions for extraction of the polysaccharide were as follows: temperature of 60℃, liquid and material in the ratio of 100 to 1, and 4 hours of extraction duration. The extraction rate of polysaccharide is up to 17.65% under such conditions. The polysaccharide can inhibit the growth of Escherichia coli and Staphyloccocus aureus, and the scavenging effects of polysaccharide on the superoxide anion radical and nitroso were significant. It could be concluded that the polysaccharide displayed obvious bioactivities.
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  Purification of polysaccharide of Phaeoporus obliquus and the bioactivity of purified fractions

  Mycosystema. 2009, 28(4): 564-570.

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  The content of polysaccharide of fruiting body and mycelia of Phaeoporus obliquus were determined and their values were 2.63% and 2.12% respectively. The methods of DEAE Sephacel column chromatography and Sephades G-200 column chromatography were employed to purify the raw polysaccharide of fruiting body and mycelia. The results showed that there were four kinds of fractions in raw polysaccharide of fruiting body, and they were marked as B1, B2, B31 and B32 and their contents were 50.20%, 32.25%, 4.48% and 6.61% respectively; fractions M1, M2 and M3 were purified from raw polysaccharide of mycelia and their contents were 48.23%, 31.85% and 10.91% respectively. Bioactivity of purified fractions were studied and the results showed that antioxidative activities were in sequence of B1＞M1＞M2＞B2＞M3＞B32＞B31. B2 and M2 could inhibit the proliferation of leukemia K-562 cells and lung cancer SMMC7721 cells in vitro; B31, B32 and M3 could reduce the blood sugar of mouse with alloxan diabetes and don’t affect the blood sugar of healthy mouse.
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  Purification and characterization of an extracellular xylanase from Thermomyces lanuginosus

  Mycosystema. 2009, 28(4): 571-576.

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  Purification and characterization of an extracellular xylanase produced by Thermomyces lanuginosus W205 was studied. The enzyme was purified to homogeneity by ammonium sulfate precipitation followed by anion exchange chromatography on Q-Sepharose FF column, with 46.6% yield and 1396.9U/mg specific activity. The optimal temperature and pH for the xylanase activity was 75℃ and pH 7.0, respectively. The xylanase was stable within pH 5.5－10. The residual activity was up to 70% after the enzyme was incubated at 70℃ for 30min. Thin-layer chromatography (TLC) analysis showed that the xylanase released mainly xylobiose and xylotriose from birchwood xylan. The hydrolysis of xylotriose appeared to proceed via transglycosylation, since the xylobiose was the predominant product. Easy purification from the fermentation broth and its good pH and thermal stability will be highly useful for its industrial application.
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  Comparative studies on lignin degradation enzymes produced by three species of white-rot fungi and combination of the strains

  Mycosystema. 2009, 28(4): 577-583.

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  Trametes cinnabarina, Pleurotus ostreatus and Phanerochaete chrysosporium are high-yielding strains producing lignin degradation enzymes. Comparative studies on lignin degradation enzymes produced by these three species of white-rot fungi and combination of the strains were conducted. The results showed that liquid-shaking culture was the best culture method, yeast extract liquid medium was the best medium. As far as laccase (lacs) production was concerned, the enzyme activities of lacs produced by the combination of Pleurotus ostreatus and Phanerochaete chrysosporium were the highest, reaching 450U/L on day 6; the enzyme activities of manganese peroxidases (mnps) produced by the combination of Trametes cinnabarina and Pleurotus ostreatus were the highest, reaching 1050U/L on day 10; the enzyme activities of lignin peroxidases (lips) produced by the combination of Trametes cinnabarina and Phanerochaete chrysosporium were the highest, reaching 2990U/L on day 8. The activities of three kinds of main lignin degradation enzymes produced by combination of the three species of white-rot fungi were much higher than those produced by single strain of the fungi. It is suggested that combined strains might be effective for increasing the yield of enzyme production.
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  Differential proteomics and enzyme changes of Hypsizigus marmoreus mycelium under cold stress

  Mycosystema. 2009, 28(4): 584-590.

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  The variation of proteome in mycelium of Hypsizigus marmoreus was analyzed under suitable low temperature by using proteomic methods and techniques for searching the different proteins related closely to cold stress and investigating the function of the proteins. Twenty-six changed protein spots were found under cold stress conditions. Changed protein spots were identified by MALDI-TOF-MS/MS, and fourteen proteins were successfully recognized. In the fourteen differential proteins, actin proteins and tubulin proteins are important components of cytoskeleton; proteasome and acetolactate synthase are involved in the synthesis and decomposition of protein; ATP synthase, transaldolase, formate dehydrogenase and RubisCO are concerned with the metabolize of energy and substance; guanine nucleotide binding protein and 14-3-3 protein participate in control of signal transcription; nascent polypeptide-associated complex may be related to the differentiation and development of cell. It is showed that there may be different kinds of proteins and enzymes involved in the control of Hypsizygus marmoreus in low temperature. The activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) raised rapidly in 9 hours of treatment and were higher than those in control, and afterward dropped gradually. Malondialdehyde (MDA) increased in 12h, and later slightly declined, indicating that the cell membrane of Hypsizigus marmoreus was injured. However, Hypsizigus marmoreus possesses strong adaptability in low temperature to a certain extend.
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  Antioxidant activity of endophytic Chaetomium globosum from Eucommia ulmoides

  Mycosystema. 2009, 28(4): 591-596.

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  Antioxidant activity of fermentation broth extracts of Chaetomium globosum strain No. 173 isolated from leaves of Eucommia ulmoides was studied. The reducing power determination, the bleaching of β-carotene suboleic acid model and the riboflavine photosensitization system were applied to evaluate the antioxidant effects of the extracts. Total phenolic content in the extracts was estimated by using the Folin-Ciocalteu colorimetric method. The results showed that the antioxidant capacity of the extracts was equivalent to Vc, and the capacity scavenging O2-? was better than rutin. The phenolic content of the extracts was 255.53±1.38mg/g (gallic acid equivalent).
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  Evaluation of the antioxidant activity of extracts from anamorphic mycelia of Cordyceps militaris in vitro

  Mycosystema. 2009, 28(4): 597-602.

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  The extracts of different solvents from anamorphic mycelia of Cordyceps militaris were analyzed for their antioxidant activities in different test systems. The results showed that all kinds of extracts have evident effects on scavenging 1,1-diphenyl-2-picrylhydrazyl radical (?DPPH) and hydroxyl radical. For the extracts of deionized water, 70% ethanol and 70% acetone at 50mg/mL, DPPH radical scavenging activity was 89.2%, 83.6% and 75.9%, respectively. When the concentration of 70% ethanol extracts was at 30mg/mL, hydroxyl radical scavenging activities was up to 100%. When the extracts of 70% acetone and deionized water were at 50mg/mL, hydroxyl radical scavenging activities was 95.6% and 89.6%, respectively. In certain concentration, the extracts contain restraining effects on pyrogallol autooxidation. Reducing power of the extracts followed in the order: 70% ethanol >deionized water >70% acetone. The concentration of the extracts is in direct proportion to the antioxidant activity, and the antioxidant components may be different in various extracts.
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  A preliminary study on the antimicrobial activities of Nectria sp. JZ6, an endophytic fungus isolated from the bulbs of Fritillaria thunbergii

  Mycosystema. 2009, 28(4): 603-609.

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  An endophytic fungus Nectria sp. was isolated from the fresh bulbs of Fritillaria thunbergii for the first time. The fungus can secrete antibacterial metabolites when cultured in CzapeK-DoX medium. In this study, the culture medium and fermentation conditions of the fungus were modified and optimized by single factor analysis and orthogonal tests. After being inoculated in the modified CzapeK-DoX medium (8% glucose, 0.5% peptone, 0.05% KCl, 0.1% K2HPO4 and 0.15% MgSO4, pH 6.5), the fungus was cultured in a rotary shaker at 150r/min and 28℃ for 6 days. The result showed that antibacterial activity of the fermentation broth of the modified medium increased by 26% as compared with that of unoptimized medium. Further analysis showed the antibacterial activity of the fermentation broth was stable at 100℃ for 30min in the range of pH1.0－5.0, and vanished beyond pH9.0. The ethyl acetate extracts from this fermentation broth could inhibit the growth of Staphylococcus aureus, Sarcina lutea, Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli in the minimum inhibitory concentration (MIC) of 0.625, 0.625, 1.25, 1.25 and 1.25mg/mL, respectively.
Papers
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  Helicogonium, a new recorded genus of mycoparasitic fungi from China

  Mycosystema. 2009, 28(4): 610-611.

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  Effect of exogenous Se on the activities of protective enzyme system and peroxidation of membrane lipoids in fruit bodies of Lentinus giganteus

  Mycosystema. 2009, 28(4): 612-615.

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short communications
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  Analysis of arbuscular fungi on remedying pesticide polluted soil

  Mycosystema. 2009, 28(4): 616-621.

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