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15 July 2011, Volume 30 Issue 4
    

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    Review
  • Mycosystema. 2011, 30(4): 515-518.
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    According to International Code of Botanical Nomenclature and Nomenclatural Code for Chinese Scientific Names of Fungi and Lichens adopted by the Mycological Society of China in 1986, the nomenclature of the following six important Chinese medicinal fungi is suggested: Taiwanofungus camphoratus (M. Zang & C.H. Su) Sheng H. Wu et al., Inonotus obliquus (Ach. ex Pers.) Pilát, Auricularia auricula-judae (Bull.) Quél., Perenniporia robiniophila (Murrill) Ryvarden, Pholiota microspora (Berk.) Sacc. and Ophiocordyceps sinensis (Berk.) G.H. Sung, J.M. Sung, Hywel-Jones & Spatafora.
  • Mycosystema. 2011, 30(4): 519-525.
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    Observation of arbuscular mycorrhizal (AM) colonization status and measurement of colonization percentage is an important fundamental work in mycorrhizal research. Methods for root staining and quantifying AM colonization percentage is reviewed, compared and evaluated in this paper. Ink-vinegar staining method observing colonization status, root-segment colonization weighting method, and magnified intersections method measuring colonization percentage, are considered to be appropriate approaches in AM studies with greater scientific, accurate and feasible values when compared with other methods. Different observation methods can be used for different purposes. For example, magnified intersection method could be chosen to observe and measure the arbuscule development status of AM fungi, whilst direct counting method could be employed to count the numbers of vesicles and entry points per unit root length. Thus the research results obtained for specific purposes are comparable. It is necessary to establish molecular techniques and fatty acid quantitative method to measure colonization status of one or more species of AM fungi, which will promote studies in physiological and ecological function.
  • Papers
  • Mycosystema. 2011, 30(4): 526-528.
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    A new species, Xylaria hainanensis, is described from dead fallen leaf in Hainan Province. Stromata of X. hainanensis are cylindrical, terminate in a sterile apex and arising in tomentose stipe. Perithecia of X. hainanensis are immersed and have conical ostiolar openings.
  • Mycosystema. 2011, 30(4): 529-535.
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    About 200 specimens of poroid wood-decaying fungi were collected from Huangshan Mountain, Anhui Province, and 85 species of polypores were identified from these materials. A checklist of the polypores is given. Substrates and collecting data of each species are provided. Among these species, Ceriporia aurantiocarnescens and C. sulphuricolor are new to China, and illustrated descriptions of the two species are given according to our materials.
  • Mycosystema. 2011, 30(4): 536-541.
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    Two polypores, Perenniporia gomezii and P. vicina, were found in Hainan Province of southern China, which are new to the Chinese mycobiota. Their illustrated descriptions are given based on the Chinese materials, and a key to accepted species of Chinese Perenniporia is presented.
  • Mycosystema. 2011, 30(4): 542-550.
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    More than 600 specimens of thermotolerant fungi were collected from 12 provinces in China, and 28 species were identified. Among them, Thielavia arenaria and Trichoderma saturnisporum are newly recorded species for China. rDNA-ITS sequencing was performed of the 28 species of thermotolerant fungi and additional 3 isolates purchased from China General Microbiological Culture Collection Center. The length of sequenced fragments were 554-895bp. Neighbor-joining phylogeny tree of rDNA-ITS sequences of the 31 isolates sequenced and 15 isolates downloaded from GenBank was used to reveal the phylogenetic relationships of thermotolerant fungi. The result of the phylogenetic analyses supports the species delimitation based on morphological characteristics.
  • Mycosystema. 2011, 30(4): 551-555.
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    The white mildew disease on Auricularia auricula-judae was epidemic in recent years in Northeast China. The pathogenic fungi formed a layer of white pull-mesh mycelium on the ventral side of the fruiting body of A. auricula-judae in hot summer. The fungi significantly reduce the yield as well as the quality of the fruiting bodies. In this study, two fungal strains were isolated from the infected fruiting bodies cultivated in Shangzhi of Heilongjiang Province. They were identified as Fusarium oxysporum and Fusarium chlamydosporum. Both of them are pathogenic to the fruiting body of A. auricula-judae.
  • Mycosystema. 2011, 30(4): 556-565.
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    Fungus-growing termites cultivate fungi on a substrate called fungus comb inside their nests or dispersed around neighborhood soil. When fungus comb is incubated without termites, fruiting bodies of the ascomycete genus Xylaria are prevailing. Four comb samples were collected from nests of fungus-growing termites belonging to the genus Odontotermes in Sichuan and Yunnan Province in southwestern China and 40 fungal strains divided into 13 morphotypes of Xylaria were isolated. Xylaria isolates were identified as two different species based on ITS1-5.8S-ITS2 sequences. To clarify the entire fungal community especially changes of the comb between a termite-activated nest and an abandoned one, ITS gene libraries were set up. Data analysis clearly demonstrated that fungus combs were monocultures of Termitomyces when fungus-growing termites live in the nest; but when they were abandoned for some time, they turned out to be polycultures of Trichoderma, Xylaria and other fungi.
  • Mycosystema. 2011, 30(4): 566-571.
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    Process of senescence in hyphae of cultivated Auricularia auricula-judae was observed microstructurally and ultrastructurally. In 30d after inoculation, the hyphae were uniform and well-knit; the cell wall was smooth. Under transmission electron microscope, the cells were intact, with abundant inclusions and all organelles were normal. In 60d part swelling hyphae were observed and the color of hyphae were darker. Under transmission electron microscope, the cell wall was unconsolidated, and the mitochondria and vacuole were swollen; nuclei were irregularly swollen, and nucleoli disappeared, and then granules and vesicles increased. Meanwhile, a few electron-dense granules appeared, which indicated the senescence of hyphae. In 90d membrane of part of the nuclei was broken, and a great amount of lipid granules, vesicles and electron-dense granules were observed. The cell wall was futher loose. In 120d lots of hyphae became fractured and the color was much darker. Under transmission electron microscope, collapsion of cell wall and disintegration of cytomembrane system were observed, and part of cell organelles, such as mitochondria, disappeared. In 150d most of hyphae were completely fractured and lost their shapes, and the cytomembrane and cell inclusions were almost disappeared, but part of collapsed cell walls were observed. It was indicated that the senescence process of Auricularia auricula-judae hyphae was irreversible, which developed from individual to whole cells gradually.
  • Mycosystema. 2011, 30(4): 572-579.
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    Blastospores are asexual fungal spores produced by budding and they are the main propagules in liquid culture for many fungi. They have been used as inoculum for solid culture for massproduction of fungal insecticides and also as an important receptor for current genetic transformation of entomopathogenic fungi. The morphogenesis of blastospores of 10 species of entomopathogenic fungi in liquid culture was studied, including Beauveria bassiana, Isaria farinosa, I. cicadae, I. cateniannulata, I. fumosorosea, I. tenuipes, I. cateniobliqua, Metarhizium anisopliae, M. acridum and Lecanicillium lecanii. The results showed two patterns of morphogenesis. I. cicadae mainly propagate as mycelium in liquid culture while the morphogenesis of the other 9 species in the liquid culture were similar and all involved two manners: yeast-like budding or hypha and/or blastospore constrictions. The second pattern included initial phase, exponential phase and senescence phase. In initial phase, blastospores were generated by budding or constriction of mycelium. In exponential phase, they were massively produced by budding or constriction of both mycelium and blastospores. In senescence phase, new blastospores were produced by blastospore budding.
  • Mycosystema. 2011, 30(4): 580-586.
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    In the process of artificial cultivation of myxomycetes, plasmodia usually could not form sporophores that resulted in the ontogeny rest on vegetative stage. In this study, plasmodia of Physarum were obtained by using liquid culture in combination with oat-agar culture, and they were induced to form sporophores. Under the condition of hunger, the optimal culture conditions of sporophore and sclerotium formation of Physarum were obtained from different media by adjusting illumination (3,000, 6,000, 9,000 and 12,000lx) and temperature (20, 22, 24 and 26℃). The optimal conditions for obtaining sporophores of P. globuliferum in oat-agar medium and liquid medium were under 24℃, 6,000lx and 20℃, 6,000lx, respectively. The optimal conditions for obtaining sporophores of P. compressum in oat-agar medium and liquid medium were under 26℃, 6,000lx, and 20℃, 6,000lx, respectively. Sporophores and sclerotia of P. melleum were all obtained in oat-agar medium, and the optimal conditions were 26℃, 6,000lx and 22℃, 3,000lx. In liquid medium, P. melleum only formed sclerotia, and the optimal conditions of were 22℃, 6,000lx.
  • Mycosystema. 2011, 30(4): 587-597.
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    17 samples of powdery mildew were collected and separated from diseased melon, cucumber, pumpkin and watermelon in greenhouses or open field in various ecological regions of Heilongjiang Province in 2010. They were investigated for physiological races by using standard differential hosts of powdery mildew. Based on their resistant or susceptible types on 13 differential hosts, three races on Cucurbitaceae were preliminarily confirmed, including races 1, race N1 and a new one of Podosphaera xanthii in Heilongjiang Province. Race 1 was dominant in Heilongjiang. Thirteen isolates of powdery mildew were evaluated by using random amplified polymorphic DNA (RAPD) with 10 primers selected from 119 random primers. A total of 157 bands were amplified, including 138 polymorphic bands with 97.89% polymorphic band frequency. The RAPD amplifying results show that high genetic diversity existed among the isolates of P. xanthii on Cucurbitaceae in Heilongjiang. After analysis by software NTSYS-PC, the similarity coefficient of the 13 isolates varied from 0.52 to 0.57. The UPGM dendrogram divided the 13 isolates into 4 groups at the similarity coefficient of 0.60. The isolates of race 1 were partly clustered into the same group, and the new race was clustered into the same group with partial race 1 isolates; two isolates of race N1 were not clustered into the same group with partial race 1 isolates. The isolates originated from the same geographic regions or hosts were also not clustered into the same group. It is indicated that no obvious association was found between DNA polymorphism and pathogen races in biological tests on cucurbits. Meanwhile, no association was found between the results of RAPD analysis and any other parameter analysis such as geographic regions, greenhouse types or host origins.
  • Mycosystema. 2011, 30(4): 598-603.
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    In order to explore early detection the occurrence of Verticillium dahliae of cotton in naturally infested soil, a SYBR Green I based Real time RT-PCR assay was developed. The positive plasmid included a 342bp PCR product was used as the reference and the specification curve was created on that basis. The specificity, sensitivity and reproducibility of the curve were evaluated and compared with conventional PCR techniques. The result showed that the Real time RT-PCR assay was rapid, highly sensitive and specific. A linear relationship was observed between the amount of input plasmid DNA and cycle threshold (Ct) values over a range of 3.8×103copies/μL to 3.8×108copies/μL, and correlation coefficient was 0.996. PCR amplification efficiency was 101.5%, which was 102 more sensitive than that of the conventional PCR.
  • Mycosystema. 2011, 30(4): 604-611.
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    Soil samples were collected in Liangshui Nature Reserve, China and enriched in medium with lignin-sodium salfacid as sole carbon source. Guaiacol-PDA plates were used to separate laccase-producing fungi. A high-yield strain NF-05 was screened after testing with 2,2′-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) and 4-hy-droxy-3,5-dimethoxybenzaldehyde azine (SGZ). NF-05 was identified as Myrothecium verrucaria based on morphological characteristics and sequence analysis of rDNA-ITS. Laccase production of NF-05 showed synchronism in concordance with hypha growth in fermentation medium. Enzyme activity reached the peak of 8,375.87U/L at the fifth day. Laccase from NF-05 was purified for assay of decolorizing azo dyes with mediators, which resulted in a decolorization percentage of 90% for methyl orange mediated by tetramethylpiperidinooxy (TE), 90.1% for orangeⅠand 100% for orange G6 mediated by TE.
  • Mycosystema. 2011, 30(4): 612-617.
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    The order Peltigerales is an important group of N-fixing lichens that are essential for ecosystem N cycling in many terrestrial ecosystems. But differences in N-fixing activity among species are not well understood at present. Twelve peltigeralean species collected in different localities and subjected to various desiccation durations, namely Collema subflaccidum, C. subnigrescens, C. tenax, Leptogium hildenbrandii, L. menziesii, L. pedicellatum, L. saturninum, L. trichophorum, Peltigera canina, P. didactyla, P. praetextata and P. rufescens, were cultured in the same condition. N-fixing activity was measured using acetylene reduction assay after 1d, 10d and 15d cultivation. No significant differences in N-fixing activity for all lichens were found between 10d and 15d cultivation, suggesting N-fixing activity for lichens was fully recovered after 10d cultivation. A significant difference in N-fixing activity among species was found in terms of mean N-fixing activity averaged over 10d and 15d cultivations (P<0.001), with the highest value for Leptogium trichophorum [(4.532±0.368)μmol C2H4/gdw·h] which is about 2 times higher than that for Peltigera canina [(2.349±0.223)μmol C2H4/gdw·h]. N-fixing activity of the other 10 lichens was roughly similar (2.635-3.379μmol C2H4/gdw·h). These results show lichens have high N fixation plasticity, which can be an evidence for the adaption of lichens to fluctuating environment conditions.
  • Mycosystema. 2011, 30(4): 618-623.
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    Five anions (F?, Cl?, NO2?, PO43?, NO3?) in Tricholoma giganteum fruiting body were determined by using ultrasonic assisted extraction (UAE)-ion chromatography (IC). After ultrasonic assisted extraction for 30min in NaOH solvent (3.0mmol/L), sample introduction as 100μL, Na2CO3+NaHCO3 (3.5mmol/L+1.0mmol/L) were chosen as washing solution with flowing rate at 1.2mL/min. The linearity ranges of 5 anions respectively were 2-25mg/L, 2-20mg/L, 2-100mg/L, 10-55mg/L, 10-100mg/L. The relative standard deviations (RSD) of the peak areas were 0.58%, 1.27%, 0.73%, 0.92%, 2.33%, respectively. The detection limits of the 5 anions were 0.0976-1.0984mg/L. The spike recoveries of 4 anions (F?, Cl?, PO43?, NO3?) were 95%-110%, whereas, the content of NO2? in T. giganteum was not detected out.
  • Mycosystema. 2011, 30(4): 624-629.
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    Analysis of heavy metal contents in Agaricus bisporus fruiting body showed the background values of lead and cadmium were 0.636mg/kg and 0.127mg/kg respectively. Lead content of the samples is in the range of 0.087-2.40mg/kg, and cadmium content 0.038-0.301mg/kg. Based on two difference cultivation methods, levels of lead in fruiting bodies have significant differences, but the content of cadmium is not significantly different. Correlation analysis showed that lead of fruiting body was mainly originated from the covering soil and had no relation with the compost. However, cadmium content in fruiting body had relation with the cultivation methods and was influenced by both compost and covering soil.
  • Mycosystema. 2011, 30(4): 630-635.
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    Three kinds of extracts were made from the fruiting body of Phellinus yamanoi by gradient extraction method using petroleum ether, methanol and water as extracting solution, and 4,6,8(14),22(23)-tetraen-3-one-ergostane was isolated from petroleum ether extract. The anti-tumor activities of the three extracts and the monomer compound against mice Hepatoma H22 were studied in vivo. Inhibition rates of tumor, immune organ indexes and the contents of immune factors were evaluated as indicators. The results showed that the inhibition rates of petroleum ether extract for high dosage group (100mg/kg) and those of monomer compound for middle dosage group (7.5mg/kg) were 62.21% and 57.67%, respectively. Spleen indexes and thymus indexes of petroleum ether extract and monomer compound were higher than those of control group and cyclophosphamide-treated group (CTX) (P<0.01), meanwhile inter leukin-2 (IL-2) level of them was obviously higher than that of the control group and cyclophosphamide-treated group (P<0.01). Tumor necrosis factor-α (TNF-α) level of petroleum ether extract and monomer compound was significantly lower than that of the control group (P<0.01). Hence, the petroleum ether extract and the monomer compound of fruiting body of P. yamanoi inhabit Hepatoma H22 cell of mice, and also can improve mice’s immune function.
  • Mycosystema. 2011, 30(4): 636-643.
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    A marine fungal strain F62 primarily determined as Aspergillus versicolor was isolated from sponge Haliclona simulans. The sponge-associated fungus was cultivated on the rice solid medium at room temperature (about 25℃) for 45 days. The culture was extracted with ethylacetate and the total extract was purified by means of silica gel, Sephadex LH-20 column chromatography, high performance liquid chromatography (HPLC), and so on. Six compounds were obtained and the structures were determined as alantrypinone (1), lovastatin (2), methylester of lactone ring-opened monacolin K (3), terrein (4), territrems B (5) and ergosterol (6), respectively, based on nuclear magnetic resonance (NMR), mass spectrum (MS) and other spectroscopic determination. Compound 1 and compounds 2-5 were first isolated from the genus Aspergillus and Aspergillus versicolor, respectively. 13C NMR spectral data of compound 3 were provided for the first time. Compound 4 was first reported to have anti-inflammatory activity in vitro.
  • short communications
  • Mycosystema. 2011, 30(4): 644-648.
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    Three species of wood-decaying fungi in the genus Hymenochaete, H. bispora, H. lenta and H. senatoumbrina are newly recorded in China. Hymenochaete bispora collected from Jilin Province is characterized by its basidia mostly having 2 sterigmata; Hymenochaete lenta collected from Inner Mongolia Autonomous Region is distinguished by its imbricate basidiocarps and allantoid basidiospores; Hymenochaete senatoumbrina collected from Xizang Autonomous Region is unique for its relatively large basidiospores in section Gymenochaete. The illustrated descriptions of these three species are provided based on the Chinese materials.
  • Mycosystema. 2011, 30(4): 649-652.
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    Three species of Pholiota collected from Inner Mongolia and Heilongjiang Province, P. alabamensis, P. flavescens and P. pseudosiparia, are reported for the first time in China. The morphological descriptions and illustrations are provided in this paper. The examined specimens are deposited in the Herbarium of Mycology of Jilin Agricultural University (HMJAU).
  • Mycosystema. 2011, 30(4): 653-657.
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    Four species of Mycena, M. adonis, M. luteopallens, M. viridimarginata and M. stylobates, new to China are reported. Morphological descriptions and illustrations of the species were given. The studied materilas are deposited at the Herbarium of Mycology of Ludong University (HMLD) and Herbarium of Mycology of Jilin Agricultural University (HMJAU).
  • Mycosystema. 2011, 30(4): 658-662.
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    A new leaf blight pathogen of Vernicia fordii was recorded in Guangxi, China. The diseased leaves show dark brown spots which then spread over the whole leaves. The premature defoliation may significantly reduce the production of tung seeds. Sixteen pure fungal cultures belonging to five taxa were obtained from diseased leaf tissues. Strain VT-04 was identified as the pathogen based on Koch’s postulates and pathogenicity test. Fruiting structures had not been observed on clinic specimens, but induction of sporulation was successfully achieved when the pathogen was cultured on water agar medium containing sterile pine needles under UV light. An integrated analysis of cultural characteristics and phylogeny of two molecular markers including the ribosomal DNA internal transcribed spacer (ITS) and RNA polymerase II second largest subunit (RPB2) revealed that the pathogen was conspecific with Neofusicoccum parvum. This is the first report that N. parvum can cause leaf disease of V. fordii.
  • Mycosystema. 2011, 30(4): 663-668.
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    15 strains belonging to 15 species of Pleurotus were used. Primer CO332F and CO332R were designed by the cytochrome c oxidase subunit 1 gene (CO1) sequence of Pleurotus downloaded from GenBank. Results of the first-step PCR amplification showed that all the strains can get one band. The 15 strains were divided into four groups by the band size. Species-specific primers were designed accordingly. Results of the second-step PCR amplification showed that each species can get one band by the species-specific primers. In conclusion, the 15 species of Pleurotus can be identified by two-step PCR in accordance with the band size and the presence or absence of the band.