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15 November 2013, Volume 32 Issue 6
    

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    Review
  • Mycosystema. 2013, 32(6): 931-946.
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    Due to morphological paucity and stasis, and phenotypic plasticity, phylogenetic relationships of fungi cannot easily be grasped based on merely morphological characters, cellular and subcellular structures, and biochemical and physiological features. New techniques, such as DNA and genome sequencing, comparative genomics and bioinformatics, have revolutionized the study of fungal taxonomy, and a large number of lineages in fungal evolution can be recognized this way, which, consequently, provides strong support for establishing new taxa at different taxonomic levels. Since 2000, at least one new subkingdom, four new phyla, seven new subphyla, nineteen new classes, nine new subclasses, and over forty new orders have been created. In the last three years, over 20 new fungal genera were published by Chinese mycologists, most of which are supported with molecular evidence. It is anticipated that a large number of new species, new genera, new families and even new higher taxa are going to be discovered and erected in the next ten years. We could take advantage of the highly rich fungal resources in China and make due contributions to the taxonomy of fungi. Meanwhile, fungal taxonomy is confronted with serious challenges. The challenges are mainly as follows: 1) studies are becoming more and more comprehensive and multi-technique based, which needs not only corresponding financial supports but also, for researchers, more all-round skills, broader background knowledge, and faster speed of renewal of knowledge; 2) the speed of description and documentation of new taxa is slow and needs to be accelerated in order to serve the increasing needs of people for recognition and utilization of species; 3) it is urgent to innovate our research schemes, and to adopt new techniques, new ideas and new strategies in order to advance taxonomy of fungi, to speed up the discovery and documentation of new taxa, and last but not least to serve the social progress and science development.
  • Papers
  • Mycosystema. 2013, 32(6): 947-952.
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    “Lingzhi” is one of the most important medicinal fungi, and it has been renowned and utilized in China for more than 2,000 years. Ganoderma lucidum was originally described for Britain specimens by William Curtis as Boletus lucidus in 1871. Patouillard first reported Ganoderma lucidum from China in 1907. Thereafter this scientific name (binomial name) has been used for the Chinese medicinal “Lingzhi” for more than 100 years. However, the recent taxonomical studies indicated the Chinese “Lingzhi” is different from G. lucidum in both phylogeny and morphology. The Chinese “Lingzhi” is an independent species, and its valid scientific name is G. lingzhi rather than G. lucidum. Ganoderma lingzhi has a wide distribution in warm temperate and subtropical East Asia, and it differs from G. lucidum by its pale yellow to sulphur yellow pore surface when fresh, the presence of melanoid bands in the context and thick dissepiments (80–120μm) at maturity, while G. lucidum has a distribution mostly in Europe, but also in northeast, northern, central and highland of southwest China, and it lacks melanoid bands in the context, and has a white to cream pore surface and thin dissepiments (40–80μm). Ganoderma sichuanense was originally described from Panzhihua of Sichuan Province, China. Based on the ITS sequence of its holotype, it differs from G. lingzhi in phylogeny. In addition, it was also recently found in Guangdong Province, China.
  • Mycosystema. 2013, 32(6): 953-958.
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    A total of 75 isolates of dematiaceous hyphomycetes belonging to 32 species in 21 genera were obtained and identified from 56 soil samples collected from different ecosystems of Chongqing (municipality directly under the Central Government). Among them, Gliomastix chongqingensis is a new species; Arthrocristula and A. hyphenata are newly recorded genus and species for China, respectively. The holotype of the new fungus, other all specimens (dried cultures) and living cultures studied are deposited in the Herbarium of Shandong Agricultural University: Plant Pathology (HSAUP). The isotype (dried culture) is kept in HMAS.
  • Mycosystema. 2013, 32(6): 959-966.
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    Blueberry twig blight was discovered in Shandong Province recently. Three isolates were obtained by single-spore separation method and inoculation experiment was carried out. Morphological observation and rDNA-ITS sequence analysis indicated that the pathogen is Phomopsis vaccinii, the anamorph of Diaporthe vaccinii. To our knowledge, this is the first report of Phomopsis vaccinii leading to twig blight of blueberry in China.
  • Mycosystema. 2013, 32(6): 967-977.
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    The infection process and ultra-structure of Penicillium digitatum on citrus fruits were observed by electron microscopy. The results showed that conidia around the wound germinated and produced germ tubes to penetrate the fruit pericarp cells through the wound 12hai (hours after inoculation) at room temperature. After then this pathogen extended into the sarcocarp cells, and gradually invaded the adjacent cells 24hai. With the invasion of the pathogen, the host cell wall started to be decomposed; the cells became plasmolysed; cell inclusions and organelles were coagulated, dark, and digested. Finally, the pericarp near the wound became soft and the hyphae grew out from the wound. The white mold appeared on the soften lesion 84hai. The lesions expanded gradually with about 3cm in diameter 96hai. About 120hai, the color of the mold changed to grayish green, and the whole fruit became rotted 144hai. The result of pectin labeling showed that the pectinase was produced by this pathogen to digest the pectin in the host cell wall, leading to cell wall looseness and, consequently, the whole fruit was rotted.
  • Mycosystema. 2013, 32(6): 978-985.
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    The process of teliospore germination, basidium development and basidiospore germination of Coleosporium phellodendri was observed with light microscope. The nucleus changes in germinating teliospore were also assayed by using iron hematoxylin staining method. The results showed that the immature teliospores contained two nuclei, and when the teliospores matured, the two nuclei would merge into one nucleus along with the elongation of teliospore body. When germination occurred, teliospores elongated and formed a septum to divide the cells into two parts. The upper parts with golden yellow protoplast developed into internal basidia and then basidiospores, while the basal transparent parts became pedicel-like. Germinating basidiospors formed a germ tube and developed a secondary spore. The germinating secondary spores normally formed hyphae. A few germinating secondary spores could repeatedly develop the tertiary spores. Both the teliospores and basidiospores could germinate in a wide temperature range from 5 to 25℃, with optimum temperature of 15–25℃.
  • Mycosystema. 2013, 32(6): 986-995.
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    Microsclerotia (MS) of Verticillium dahliae are main survival structure in soil and primary inoculum for Verticillium plant wilt, which may remain viable for up to 14 years in the absence of a host. The occurrence of Verticillium plant wilt was determined by the number and survival status of MS in soil. The optimal conditions for MS germination and lethal temperature were investigated by using V. dahliae strain XJ2008 as experiment material. Results showed that 20℃ and pH8.0 were the optimal conditions for MS germination. MS of V. dahliae can tolerate high temperature. The germination rate of MS showed a downward trend with the extension of treat time under different temperature. The MS germination rate declined greatly when at and above 55℃. However, it declined slowly at and below 50℃. The MS were completely dead when treated at 55℃ for 360min. There was still a small amount of MS survival when treated at 40℃, 45℃, and 50℃ for 1,440min, but their germination declined gradually with time. The simulation results of MS in soil showed that soil temperature had a strong lethal effect to MS. The MS were completely lost vital when treated in soil at 40℃ for 4 days. The results provided a theoretical base for management of Verticillium wilts across-the-board range of crops through applying plastic film to increase soil temperature.
  • Mycosystema. 2013, 32(6): 993-1003.
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    Glomalin-related soil protein (GRSP) is a glycoprotein secreted from the hyphae of arbuscular mycorrhizal (AM) fungi, and it can promote the formation of soil aggregates. Production of GRSP is affected by many factors. It is presently not clear if host root morphology affects GRSP production through competition for carbon source. In this study, two ecotypes of Glomus mosseae and red clover (Trifolium repense) were taken as biological materials to investigate the relationship between root morphology and GRSP production in sand culture. The test results indicated that AM inoculation increased the proportion of fine roots, while decreased the proportion of coarse roots, but did not affect the total root length and root surface area. Colonization of AM fungi and hyphal length increased along with time extending, and showed significant difference between the strains. Inoculation of AM fungi increased the GRSP production. The correlation analysis indicated that GRSP production was not related to root morphology, but significantly correlated with AM fungi colonization and hyphal length. It is suggested that the establishment of root morphology didn’t inhibit GRSP production although the AM fungi and host plant competed for carbon source. A possible auto-regulation mechanism might be present for the partitioning carbon source in mycorrhiza.
  • Mycosystema. 2013, 32(6): 1004-1011.
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    The Myxomycetes is a unique class of fungi. Until now, the systemic studies of myxomycetes has been mainly based on the morphological characteristics, while the molecular phylogenetic analysis has mainly been inferred from small subunit rRNA and elongation factor 1-α gene sequences. In this study, actin gene and β-tubulin gene sequences were analysed for determining if they can be effectively applied to the study on myxomycete phylogeny. Seven actin gene sequences and seven β-tubulin gene sequences were respectively obtained from field-collected fruiting bodies of eight myxomycete species. The fourteen sequences were firstly reported. Prior to this paper, available data of actin gene and β-tubulin gene sequences were quite limited because there was not any other actin or β-tubulin gene sequences of Myxomycetes in GenBank except those of Physarum polycephalum. The results showed that actin gene can effectively dividing the Liceales, Trichiales, Physarales and Stemonitales into four branches, and the order Stemonitales displayed alone in an evolutionary branch. The results supported the viewpoint proposed according to the formation of fruiting body, i.e., the class Myxomycetes had historically consisted of two phylogenetic routes. This also suggested that actin gene have an important significance in phylogenetic analysis of Myxomycetes.
  • Mycosystema. 2013, 32(6): 1012-1019.
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    Entomopathogenic hyphomycete has long been widely applied in biocontrol of insect pests due to easy production, formulation and environmental safety. This study was to illustrate the relationship between the oxidative tolerance and the polysaccharide content in aerial conidia of Beauveria bassiana and Isaria fumosorosa. Survival indices of 11 isolates were separately assessed as a ratio of the viability of conidia exposed to 10min stress with H2O2 over that of unstressed conidia and fitted well to a survival model (r2>0.95). For a given isolate, the fitted model generated an LD50, the concentration for 50% viability loss under the stress. As a result, I. fumosorosea was more tolerant to oxidative stress than B. bassiana. The LD50 for B. bassiana and I. fumosorosea was correlated well to the conidial polysaccharide content of the respective strain. The conidial contents of B. bassiana could be modified by changing the concentration and type of sugar in media. However, the survival indices of isolate were still correlated well to the polysaccharide content from conidia produced on glucose-, sucrose-, or starch-based media. The relationship between conidial polysaccharide content and oxidative tolerance suggested a new way to search for fungal biocontrol agents with more environmental stability.
  • Mycosystema. 2013, 32(6): 1020-1027.
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    The objective of this study was to evaluate the effects of harvest maturity on the aroma components of Tuber indicum and analyze the key odor compounds of ripe Tuber indicum. The truffles with different harvest maturities were subjected to solid-phase micro-extraction (SPME) and evaluated by gas chromatography and mass spectrometry (GC-MS) for aroma components. The results showed that 4 aroma compounds were identified in unripe fruiting body, 8 in half ripe one and 13 in ripe one. Most of aroma compounds from ripe stage were previously described as responsible for truffle aroma. By calculating the odor activity value (OAV) of each aroma compound in ripe fruiting body, it was found that dimethyl sulfide, 2,3-butanedione, 3-methylbutanal, 2-methylbutanal, hexanal, and 1-octen-3-ol were the key odor compounds of Tuber indicum (OAV>1).
  • Mycosystema. 2013, 32(6): 1028-1033.
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    Two novel natural products, 2-(ethoxymethyl)-3,5-dimethoxybenzyl acetate (1), 2-(acetoxymethyl)-4, 6-dimethoxybenzyl tridecanoate (2), and two known compounds, 5,7-dimethoxyisobenzofuran-1(3H)-one (3), 4,6-dimethoxyisobenzofuran-1(3H)-one (4) were isolated from the solid culture of Pleurotus cornucopiae. Compounds 3 and 4 showed moderate cytotoxicity against HeLa cell line with the IC50 values of 66.2 and 65.7μmol/L, respectively. Compounds 3 and 4 inhibited NO production in LPS-activated murine macrophage RAW264.7 cells with the IC50 values of 17.2 and 67.9μmol/L, respectively.
  • Mycosystema. 2013, 32(6): 1034-1045.
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    In order to set up the basic process for naringinase production by solid-state fermentation and to evaluate its practical value, the authors have studied the fermentation of pomelo peel to produce naringinase from Aspergillus aculeatus JMUdb058. The naringin hydrolysis, naringinase and α-L-rhamnosidase activities were detected by a high performance liquid chromatography method. Soybean meal and the medium sterilization were investigated for their effects on naringinase production by single factor experiments. In addition, the dynamic trends of several fermentation parameters, including naringinase, α-L-rhamnosidase, reducing sugar and biomass, were analyzed. Furthermore the naringinase was evaluated in the debittering efficiency by monitoring the naringin hydrolysis in pomelo juice. The results showed that the solid-state fermentation of pomelo peel could produce naringinase, and the addition of soybean meal could significantly increase the activity of naringinase. However, sterilization of the medium (121℃, 20min) could seriously decrease the naringinase production. A medium consisting of pomelo peel powder (5g), soybean meal (0.75g) and H2O (5.75mL) which was inoculated and kept for cultivation at 30℃ for 8d had the highest productivities of naringinase (5.81IU/gds) and α-L-rhamnosidase (6.08IU/gds). During the fermentation, both the naringinase and α-L-rhamnosidase rapidly synthesized in the logarithmic grown phase, and its kinetic model showed that the syntheses of those two enzymes were strictly associated with the growth. Moreover, 99.6% of the naringin in the pomelo juice was hydrolyzed by naringinase, indicating its high effectiveness in debittering citrus juice. These results suggest that pomelo peel is a good material for naringinase fermentation, and that solid-state fermentation of pomelo peel is a high-efficiency way to produce naringinase from A. aculeatus.
  • Mycosystema. 2013, 32(6): 1046-1055.
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    The inhibitory effects of dry matter of culture broth (DMCB) from Phellinus igniarius submerged fermentation and its fractions were evaluated with the mice transplanted Hepatoma 22 in vivo. The results showed that both of DMCB and its fractions exhibited anticancer effect and increased the life span. The potency of DMCB and its fraction II (polysaccharides) seems to be better than that of the other two fractions. The inhibitory rate at the high dose of DMCB (1,000mg/kg/d) and fraction II (360mg/kg/d) was 33.5% and 40.3% respectively. The histopathological results showed that necrosis in H22 cells was induced in both DMCB and fraction II treatment groups, and immunohistochemical detection indicated the decrease in Bcl-2 expression and the increase in Bax expression in the cancerous tissue of the mice. DMCB from Phellinus igniarius submerged fermentation and its polysaccharides fraction possess potent antitumor activity in vivo.
  • Mycosystema. 2013, 32(6): 1056-1063.
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    Anti-tumor activities against H22 tumor and the immunopotentiation to mice of Ganoderma lucidum-fermented product of ginseng-simmered extract were evaluated in five aspects, viz. inhibition rates of tumor, immune organ indexes, nonspecific immunity, the humoral immunity and the cellular immunity. The inhibition rates of the fermentation products for high dosage treatment group reached 51.65%. The spleen indexes and thymus indexes were higher than those of the control group and cyclophosphamide-treated group (CTX). In the immunological enhancement experiment, the medicine treatment groups had significant difference as compared with the control groups (P<0.01). These results indicated that Ganoderma lucidum-fermented product of ginseng-simmered extract could suppress tumor growth and had potent enhancement effect on the immune function of mice significantly.
  • Mycosystema. 2013, 32(6): 1064-1070.
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    A method for determination of seven organic acids in edible fungi using reversed-phase high performance liquid chromatography (HPLC) was improved. The HPLC system was as follow: Green ODS-AQ column was used at 30℃ and mobile phase was 10mmol/L KH2PO4 (pH2.8) with flow rate of 1.0mL/min. The UV detection wavelength was 210nm and sample volume was 10μL. The results indicated that seven kinds of organic acid RSD value were less than 5% in precision, repeatability and stability experiment of this method, and sample recovery rate was between 94.6% to 99.3%. The method was simple which could be applied to determining organic acids in edible fungi. Seven organic acids in eight kinds of mushrooms were analyzed under favorable conditions. Results showed that the variety and content of organic acids in these mushrooms are quite different.
  • Mycosystema. 2013, 32(6): 1071-1078.
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    Three methods (cotyledon in vitro refrigeration, leaf in vitro refrigeration and 10% dimethyl sulfoxide plus 5% skimmed milk refrigeration) were used to preserve sporangia of Peronospora parasitica, the pathogen of Chinese cabbage downy mildew. Cotyledon in vitro inoculation method was adopted to measure the pathogenicity of sporangia of P. parasitica for screening a better preservation method. The experimental results indicated that the sporangia preserved in 6 months by use of cotyledon in vitro refrigeration method and leaf in vitro refrigeration method had higher germination rate, incidence and disease index, on the contrary, when the sporangia were preserved by use of 10% dimethy sulfoxide plus 5% skimmed milk refrigeration method, germination rate, incidence and disease index have decreased to 2.16%, 0 and 0 respectively. Cotyledon in vitro refrigeration method showed best preservation effect. After being preserved for 12 months, germination rate, incidence and disease index of the pathogen were 62.22%, 90% and 48.89 respectively. The sporangia preserved by use of leaf in vitro refrigeration method showed weak pathogenicity, and the sporangia lost their vitality when they were preserved by use of 10% dimethy sulfoxide plus 5% skimmed milk refrigeration method. Thirty-two isolates of the pathogen were successfully rejuvenated by 93.75% when they were preserved by use of cotyledon in vitro refrigeration method in 12 months. It is concluded that cotyledon in vitro refrigeration is a good method for long period preservation of P. parasitica with higher germination rate.
  • short communications
  • Mycosystema. 2013, 32(6): 1079-1085.
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    During June to October, 2010, we investigated and collected freshwater fungi from Shandong Province. Three new Chinese records were identified: Hyalocamposporium longiflagellatum, Spencermartinsia uruguayensis and Pleospora pelagica. In this paper, these species are described, illustrated and discussed.
  • Mycosystema. 2013, 32(6): 1086-1096.
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    Three hydnaceous wood decaying fungi, Steccherinum ciliolatum, S. oreophilum and S. robustius, were newly founded from China. They were collected from Heilongjiang, Sichuan and Jilin provinces respectively. Both the morphology and phylogenetic analysis of nLSU sequences support these three new records. S. ciliolatum is characterized by resupinate basidiocarps, cream to buff hymenial surface, margin fimbriate, short aculei that are less than 0.5mm and ellipsoid spores. S. oreophilum is easily recognized by its small, effuse-reflexed basidiocarps with glabrous pileus and irpicoid to poroid hymenial surface, subulate skeletocystidia and narrowly ellipsoid spores. S. robustius is characterized by resupinate to effused-reflexed basidiocarps, reddish-orange to dirty-yellow spines and broadly ellipsoid spores. Phylogenetically, these three new species form well-supported, terminal monophyletic clades, distinct from other sampled Steccherinum species. Illustrated descriptions of these three species are given based on the materials from China. A key to the accepted species of Steccherinum in China along with a synoptic description of each species is provided.