15 January 2011, Volume 30 Issue 1

    

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Review
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  A glance back at the thirty years’ Mycosystema and prospect

  Mycosystema. 2011, 30(1): 1-4.

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  The present paper deals with the following three points: 1. A brief history of starting publication of the mycological journal in China. The first issue of the Acta Mycologica Sinica in Chinese was published in 1982. The first issue of the Mycosystema in English was published in 1988, which as one of the 25 periodicals in mycology of the world was listed by the Dictionary of the Fungi (8th Edition). In this case, the Acta Mycologica Sinica was merged into the Mycosystema in 1997. And the volume ordinal numbers of the Acta Mycologica Sinica was continued to use for that of the merged Mycosystema. 2. The three storage and retrieval systems of bioinformatics (publications), culture collections and herbaria of pan-fungal specimens as prototype references of the pan-fungal taxonomy are the most important support system to biological resources. 3. The Mycosystema as one of the three storage and retrieval systems played the leading role in the academic exchange of the mycology at home and abroad. The author hopes to see the Mycosystema will play more important role in the academic exchange of the mycology.
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  The diagnosis of invasive fungal diseases: current status and future perspectives

  Mycosystema. 2011, 30(1): 5-11.

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  During the last 20 years, significant progress had generated in numerous regions of medical science. However, the worldwide incidence or mortality of invasive fungal diseases (IFDs) are still increasing remarkably, and being a serious threat to human health. Absence of typically clinical manifestations and optimal therapy options of IFDs lead to high mortality and morbidity of these infections worldwide. So earlier and specific diagnosis can distinctly improve the prognosis of IFDs. Despite the limitations of the morphological methods, represented by culture or pathology, they are still the gold standard of diagnostic invasive fungal diseases at present. The serological and imaging test such as G/GM test or high resolution CT deserved to be vigorously promoted in the clinical diagnosis; Although its clinical application is still long way to go, nucleic acid diagnostics based on PCR technology is likely to have a bright perspectives. Currently, combination and improvement of the current methods such as morphological methods, serological methods and advanced imaging technology is the best and most realistic approach to the diagnosis of IFDs in clinics.
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  Research advances of fungal diversity in combs of fungus-growing termites

  Mycosystema. 2011, 30(1): 12-17.

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  Fungus-growing termites cultivate fungi on a substrate called ‘fungus comb’ inside their nests. Fungus comb has rigid, friable and porous structure, which is considered as a unique fungus-living environment. When fungus-growing termites live in the nest, the mutualistic basidiomycete Termitomyces species are the dominant fungi on the comb; when the termite colony dies, or fungus comb is incubated without termites in the laboratory, fruiting bodies of the ascomycete genus Xylaria appear and rapidly cover the fungus comb. There are other microbes such as anamorphic fungi and yeasts living on the comb. Many of the fungi on the comb have great underlying medicinal and economic values which are worthy of studying. In this paper, the research advances in fungal diversity of the fungus comb are summarized based on several primary fungal groups like Termitomyces, Xylaria etc. Recent molecular ecology methods used to study the fungal community of the comb are discussed. Besides, the paper reveals current research hot issues and existing problems, as well as further possible solutions.
Papers
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  Diversity of endophytic fungi from Pteroceltis tatarinowii

  Mycosystema. 2011, 30(1): 18-26.

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  Diversity of endophytic fungi from Pteroceltis tatarinowii was studied in order to understand diversity and succession change of endophytic fungal communities in this endemic plant. Healthy plant samples were collected from five natural distribution areas of P. tatarinowii in Anhui, Henan, Shandong and Jiangsu Provinces. A total of 3,611 isolates of endophytic fungi were obtained. Thirty-one genera belonging to Ascomycota (2.25%), Coelomycetes (49.42%) and Hyphomycetes (28.52%) were identified, of these Phomopsis (22.10%) and Alternaria (20.66%) were the dominant genera. A total of 19.52% of all isolates were sterile on the media. The community of endophytic fungi from P. tatarinowii of Nanjing and that of Zaozhuang has the highest similarity (Cs = 0.84), while the community similarity of Nanjing and that of Nanyang was the lowest (Cs = 0.60). Coelomycetes was the most dominant in lamina, petiole and branch. The isolation rate of Hypomycetes was the highest (46.71%) in spring, but that of Coelomycetes (55.85%) was the highest in autumn.
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  The pathogen causing mango malformation in Jinsha River Dry-hot Valley, China

  Mycosystema. 2011, 30(1): 27-31.

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  Mango tissue samples infected by malformation disease were collected from Jinsha River Dry-hot Valley and the fungal strains were isolated after surface sterilization. Isolate MG6 was confirmed to be the pathogen of this disease through Koch’s rules. Based on morphological characteristics and EF-1α sequencing, isolate MG6 was identified as Fusarium mangiferae. The colony of the strain was white and pigment was not formed on Potato Sugar Agar (PSA). Pigmentation on rice medium was pink. On Carnation Leaf Agar (CLA) medium, microconidia formed false head on branched monophialides or polyphialides, and conidia in chains were not observed. Microconidia are ovate to long-oval, 0－1-septate, 3.1－10.2×1.5－2.2μm. Macroconidia are falculate, 3－5-septate, 18－38×1.8－2.4μm. Chlamydospores are absent. The EF-1α region of the pathogen was amplified and sequenced. A blast search through the Fusarium database (http://isolate.fusariumdb.org) showed that the identity of EF-1α sequence of isolate MG6 to that of Fusarium mangiferae NRRL 25226, the closest match was 99.68%, confirming the morphological identification of the isolate.
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  A comparison of a bunt smut from imported bluegrass seeds with its allied fungal species

  Mycosystema. 2011, 30(1): 32-38.

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  Several spicies of the bluegrass (Poa L.) are important herbage and lawngrass. In recent years, an undetermined species of Tilletia (T. sp.) which causes bunt smut of Poa spp. had been frequently intercepted from introduced seeds of Kentucky bluegrass, Canada bluegrass and annual bluegrass. An integrated comparison of the spicies on morphological, germination and cytological characters of teliospores with several morphologically allied Tilletia species, T. sterilis, T. togwateei, T. bromi and T. fusca was carried out. The result showed that there are significant discrepancies between T. sp. and T. sterilis, T. togwateei, while the differences between T. sp. and T. bromi, T. fusca are not obvious. The T. sp. is more closer to T. bromi morphologically. According to the results, we considered that the T. sp. and T. bromi are same species.
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  Construction of rDNA-mediated stable expression vectors in Saccharomyces cerevisiae

  Mycosystema. 2011, 30(1): 39-45.

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  A fragment containing ori of ColE1 and bla gene was inserted in the EcoR I or Hpa I site of the rRNA gene segment from Saccharomyces cerevisiae, respectively. Based on the expression vector pYES2 and the above two recombinant fragments, two novel rDNA-mediated stable expression vectors, designated as pHBM367E/H, were constructed. The glucoamylase gene from Aspergillus niger was inserted into the vector pHBM367H and the resultant plasmid was named pHBM166. For bio-safety consideration, the plasmid pHBM166 was linearized by Hpa I digestion to remove ColE1 ori and bla gene. Digested product was transformed into S. cerevisiae Y33. Transformants showed different expression levels of glucoamylase. Stability research showed that the selected transformant can express glucoamylase stably in S. cerevisiae for at least 80 generations.
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  Transformation of Flammulina velutipes with a basic fibroblast growth factor (bFGF) by Agrobacterium tumefaciens

  Mycosystema. 2011, 30(1): 46-53.

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  A bFGF expression vector p139035S-bFGF was constructed and trasnsformed into Flammulina velutipes via Agrobacterium tumefaciens using a hygromycin phosphotransferase gene (Hyg) as a selective marker. The lowest inhibitory concentrations of hygromycin for F. velutipes were found to be 9μg/mL on PDA solid medium and 6μg/mL in liquid medium, respectively. The toxicity of cefotaxime to F. velutipes and the critical factors influencing transformation efficiency including the concentration of bacterium, infection time, the concentration of acetosyringone (AS), and the time of co-culture were tested. Our result showed that cefotaxime had no negative effect on F. velutipes growth and the best selective concentration was 400μg/mL. The maximum transformation efficiency was achieved under a condition when the bacterial growth reaches an OD600 value of 0.5, infection time was about 30min, AS concentration was 200μmol/L and co-cultivation time was 72h. The results of PCR and Southern blot indicated that bFGF was integrated into the F. velutipes genome. After five successive transfers on solid PDA medium without selection pressure, the bFGF gene was still detectable in transgenic F. velutipes strains, suggesting the stable integration of the exogenous gene.
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  Molecular genetic structure of the B mating-type locus of Lentinula edodes II. Molecular organization of the B mating type locus of a L. edodes monokaryon

  Mycosystema. 2011, 30(1): 54-59.

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  In our previous work, degenerate PCR and chromosome walking technologies were used to obtain one pheromone receptor gene and one pheromone precursor gene from Lentinula edodes. In this study, four pairs of specific primers were designed according to the whole genome sequencing of the protoplast monokaryon of Lentinula edodes strain 135, to amplify STE3-like pheromone receptor gene and its flanking conserved genes in the protoplast monokaryon strain SUP2 derived from Lentinula edodes strain Suxiang and 33,655bp DNA sequence was obtained. By BlastX search, seven putative genes were identified, and three of them are pheromone receptor encoded genes. Furthermore, near to two pheromone receptor genes, four genes encoding proteins with conserved motifs of pheromone precursors were found. This study firstly reveals the molecular organization of the B mating type locus of Lentinula edodes.
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  Screening and breeding of lipase-producing fungi and a study of the lipase properties

  Mycosystema. 2011, 30(1): 60-68.

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  Penicillium chrysogenum J23 lipase activity reached maximal level of 6.40U/mL. Maximum enzyme production was observed in the medium containing sucrose 0.1%, peptone 2.0%, olive oil 1.0%, and MgSO4·7H2O 0.05%. The medium was inoculated with an initial volume of 1.0% seed culture, agitated at 200r/min and incubated for 48h. The optimal temperature and acidity for lipase to decompose lipid was found at 30℃ and pH9.0, respectively. Lipase characterization showed that its maximal activity was observed at 33℃ and pH7.5. Moreover, the lipase was highly stable under the pH value ranging from 6.0 to 10.0, and indicated 30% of total activity at 50℃. The results also showed that this kind of lipase was activated by high concentration (50mmol/L) of common detergent ions Ca2+, Mg2+, K+, but inhibited by Fe2+, Mn2+, Cu2+, Pb2+, Li2+.
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  Fermentation culture of Naematoloma fasciculare and antitumor activities of its fermented broth in vivo

  Mycosystema. 2011, 30(1): 69-76.

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  Optimum conditions of fermentation culture of Naematoloma fasciculare were investigated by using central composite rotatable design. Antitumor activities of fermented broth of N. fasciculare were observed on H22 implanted mice in vivo. The results showed that the maximal biomass was 15.06g/L when the composition of the medium was 59.27g/L maltose, 8.04g/L peptone:yeast powder=1:1 and 2.27g/L KH2PO4:MgSO4:(NH4)2SO4=1:1:1. There was an obvious dose-effect relationship between dosages of fermented broth of N. fasciculare and antitumor rates, and the maximum inhibition rate reached 54.71% at concentration of 200mg/kg. In addition, WBC (109/L) and content of IL-2 of the mice in the group at concentration of 200mg/kg were all improved obviously as compared with control group.
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  Protective effects of crude polysaccharides from Chroogomphus rutilus on dopaminergic neurons impaired by MPTP in mice

  Mycosystema. 2011, 30(1): 77-84.

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  To investigate the protective effects of crude polysaccharides from Chroogomphus rutilus on doaminergic neurons impaired by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in C57BL/6J mice were investigated, and to analyse the antioxidant activity of the crude polysaccharides in vitro was analysed. The mouse model of Parkinson’s disease was duplicated by using intraperitoneal injection of MPTP. The mice were pretreated through gavage with different dosages of crude polysaccharides (100mg/kg, 200mg/kg and 400mg/kg/day) from Chroogomphus rutilus and then the behavior and the quantitative changes of substantia nigra dopaminergic neurons were observed in these mice by behavioral detection and immunohistochemical methods. The antioxidant activity of the crude polysaccharides was measured by using the method of inhibition of lipid peroxidation in mice. The results showed that pretreatment with crude polysaccharides from Chroogomphus rutilus at dosages of 200mg/kg and 400mg/kg significantly improved the behavioral symptoms of the mice with Parkinson’s disease and thereby effectively protected dopaminergic neurons of the mice from apoptosis induced by MPTP. The protected effects may relate to the antioxidant activity of polysaccharides. This indicates that the polysaccharides from Chroogomphus rutilus are one of the possible candidates for drug discovery to treat Parkinson’s disease.
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  A preliminary study of fresh Hericium erinaceus oral liquid on health-care function and promotion of learning and memory

  Mycosystema. 2011, 30(1): 85-91.

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  The study was designed to investigate the toxicity of fresh Hericium erinaceus oral liquid and its treatment effects on learning and memory, DTH reaction in a mouse model of immunosuppression, and STZ-induced diabetes in a rat model. The results showed that the LD50 of fresh Hericium erinaceus oral liquid is higher than the dose of 27.2mL/kg. In the stay-down and stay-through assays, the time of latency and the number of errors made in the 5 minutes in the oral liquid-treated group was apparently less than those of the other groups. In the delayed type hypersensitivity (DTH) reaction, the swell of left back toes in the oral liquid-treated immunosuppressed mice was significantly bigger than that in the non-treatment immunosuppressed mice. In addition, the body weight of STZ-induced diabetic rats in the oral liquid-treated group was significantly improved in comparison with that before taking the oral liquid, and the haematological traits of diabetic rats in the oral liquid-treated group were not obvious as compared with those in the normal control group. The conclusion is that fresh Hericium erinaceus oral liquid is no toxicity, and it can significantly improve the ability of learning and memory, and strengthen the responses of the cellular immunity in mice. Besides, it can protect rats against STZ-induced diabetes.
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  Extraction conditions of glomalin-related soil protein

  Mycosystema. 2011, 30(1): 92-99.

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  Glomalin is a soil glycoprotein produced by arbuscular mycorrhizal (AM) fungi, presently named as glomalin-related soil protein (GRSP) due to its unspecific extraction. The extraction conditions of GRSP can affect its measured value, therefore, the influences of pre-sterilization of soil sample, centrifugation and soil sample mass on the measured value of GRSP were studied with citrus orchard soil. The results showed that both pre-sterilization of soil sample and centrifugation at higher centrifugal force (15,000′g) significantly affected the measured value of GRSP, indicating in the range of 0.25－1.0g the greater of soil sample mass, the higher of the measured value showed. The cumulative contents of total GRSP exceeded 66.8% and 92.1% of the theoretical maximum, respectively. These results provide a basis for the standardization of GRSP measurement, and also contribute to the comparison among the measured values of GRSP in literatures.
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  The methods for rapid DNA extraction from teliospores of Puccinia xanthii f. sp. ambrosiae-trifidae

  Mycosystema. 2011, 30(1): 100-107.

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  The genomic DNA of Puccinia xanthii f. sp. ambrosiae-trifidae was extracted by nine methods from the teliospores. The results show that the methods of CTAB-steel balls, modified mini-drill and modified EZ-Kit were better than the other six methods due to their higher quality of DNA extraction with less impurity. The template DNA samples generated from these methods were detected by methods of ITS-PCR and ISSR-PCR with the primer UBC#835. They all produced expected band patterns. The research confirms that the DNA samples generated from these three methods could be used for further molecular study of the rust fungus.
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  Effect of protoplast regeneration technology on the virus-elimination of Pleurotus ostreatus

  Mycosystema. 2011, 30(1): 108-115.

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  Three dsRNA bands with sizes of 8.2kb, 2.6kb and 1.1kb respectively, were isolated from two Pleurotus ostreatus strains, Xin-831 and Yu-6. Three virus-eliminated methods, i.e. hyphal tips isolation virus-elimination (HTIVE), primordium tissue isolation virus-elimination (PTIVE) and protoplast regeneration virus-elimination (PRVE) were applied to prepare virus-eliminated strains, and the effect of virus-elimination was detected by dsRNA technique. The results showed that no dsRNA remained in the strains by PRVE method. However, 3 dsRNA bands still remained by PTIVE method and one dsRNA band of 2.6kb, remained by HTIVE method. The results also showed that the hyphal growth rate, biomass, breath intensity and cellulose activity of virus-free strains by PRVE method were obviously superior to the original strains and strains by HTIVE and PTIVE methods. In addition, the biological efficiency of virus-free strains was 96.4% to 99.1%, and 19.9%－25.4% higher than the original strains. Moreover, the length and width of pileus increased. In all, these results indicated that protoplast regeneration technique could effectively eliminate virus of the strains and improve yield of P. ostreatus.
short communications
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  Three new records of thermotolerant fungi from China

  Mycosystema. 2011, 30(1): 116-122.

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  Three new Chinese records of thermotolerant fungi are reported, they are Chaetomium strumarium, Thielavia subthermophila and Hamigera avellanea. Illustrated descriptions of the species were given according to our strains, and their taxonomy was discussed. Specimens and living cultures examined are deposited in the Herbarium of Shandong Agricultural University, Plant Pathology (HSAUP).
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  Three Chinese new records of freshwater fungi

  Mycosystema. 2011, 30(1): 123-127.

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  Freshwater fungi Pseudomassaria sepincoliformis, Quaternaria dissepta and Fusariella sarniensis were new Chinese records. Descriptions, illustrations and discussion of the species were given. The specimens were deposited in mycology lab at Qingdao Agricultural University.
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  First discovery of Inonotus rickii on Gleditsia sinensis

  Mycosystema. 2011, 30(1): 128-132.

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  Macroscopic and microscopic observations and ITS rDNA sequence analysis were made for identifying a new pathogen of Chinese honey locust Gleditsia sinensis. The parasite was identified as Inonotus rickii. Inonotus rickii is closely related to I. patouillardii and I. quercustris.
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  Endophytic fungi producing flavonoids from Spiranthes sinensis

  Mycosystema. 2011, 30(1): 133-137.

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  Thin-layer chromatography (TLC) analysis indicated that three strains S-1, S-2, and S-3 of endophytic fungi isolated from roots, stems and leaves of Spiranthes sinensis could produce flavonoid. To identify these 3 strains, ITS DNA regions of 3 strains were amplified through PCR reaction and sequenced. The phylogenetic tree was constructed by the software MEGA (version 4.0) using Neighbour-joining methods. Morphological observation and molecular biology identification indicated that the strain S-1 was Curvularia inaequalis., and S-2 and S-3 were Penicillium pinophilum.
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  Ultrastructural studies on sporulation of Physarum compressum

  Mycosystema. 2011, 30(1): 138-141.

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  Phaneroplasmodium of Physarum compressum was induced to form fruiting body and ultrastructure during sporangium development was observed. All plasmodia participate to form spores and capillitium. The density of protoplasm and the number of lipid droplets increase during plasmodium accumulation. During spore development, the plasmodia were divided by network of vacuole associated. Spore and capillitium developed simultaneously. Primary walls of neighbouring spores were protuberant and sunken inosculatedly, forming surface ornamentation patterns of spore. The spore wall moved to appropriate location with sporangium development, and the wall of primary spore consists of two layers: an electron dense outer layer and a translucent inner layer. Round spore subsequently forms external verrucae and internal lipid droplets.
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  Mycelium cultivation optimization of Tricholoma giganteum

  Mycosystema. 2011, 30(1): 142-146.

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  Different nutritions for growth of Tricholoma giganteum mycelium were studied. The results showed that fructose was the best carbon source, and the best nitrogen source was yeast powder; the mineral element which can accelerated the growth of the mycelium was potassium dihydrogen phosphate; the growth adjustment factor was indole-3-acetic acid; the optimum temperature was 28－32℃; and the optimum pH was 4.0－5.0.
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  An efficient extraction method of pathogenic fungus DNA for PCR

  Mycosystema. 2011, 30(1): 147-149.

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  The fungal rDNA-ITS sequence analysis is suitable for systematic studies of higher level biological populations. However, this traditional method of extraction of fungal genomic DNA is procedurally overelaborate and time-consuming. Chelex-100 was adopted to extract DNA from pathogenic fungi, and the qualities of PCR amplification and rDNA-ITS sequence analysis were evaluated. The results showed that this improved method is simple, high efficient and convenient for pathogenic fungal DNA extraction.