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15 September 2010, Volume 29 Issue 5
    

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    Review
  • Mycosystema. 2010, 29(5): 625-628.
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    Publications dealing with Chinese fungal species were systematically investigated, and 2,849 new species, 129 new varieties and 5,260 new Chinese records have been reported during 1978 to 2010. “Sylloge Fungorum Sinicorum” was published in 1979, listed 6,737 species and 168 varieties based on the reports on Chinese fungal resource until 1973. So 14,846 species and 297 varieties have been found in mainland China by 2010. In addition, 2,122 fungal species were recorded in Hong Kong and 6,207 in Taiwan, among which around 800 and 400 species, respectively, were not found in mainland China so far. Until now there are 16,046 species and 297 varieties have been recorded in the Chinese territory. If 10% of them are the synonyms, the Chinese fungal species are around 14,700. Among them around 300 species are Chromista, 340 are Protoza, and 14,060 are Fungi.
  • Mycosystema. 2010, 29(5): 629-635.
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  • Papers
  • Mycosystema. 2010, 29(5): 636-643.
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    The pathogen causing hull-less pumpkin (Cucurbita moschata) powdery mildew in Wuwei District of Gansu Province was identified as Podosphaera xanthii based on the morphology of cleistothecia and conidia, conidial germination and host range, etc. The results show that the optimum temperature, relative humidity and illumination for formation of cleistothecia on hull-less pumpkin were 20℃, 70% and 4,400lx, respectively. The host range test shows that the pathogen could infect many species of Cucurbitaceae except Luffa cylindrical. Triticum aestivum, Capsicum annuum, Lycopersicon esculentum, Trifolium repens, Medicago sativa, Phaseolus vulgaris (‘Jiadouwang’) and Phaseolus vulgaris couldn’t be infected. The pathogen could also attack Phaseolus radiatus, Vigna angularis and Helianthus annuus. The most susceptible plants were Cucurbita moschata, Phaseolus radiaus, Cucurbita pepo, Vigna angularis, Helianthus annuus, Cucumis melo and Cucumis sativus, their disease incidence rates reached 100%, and the disease index were 15.56, 14.51, 13.33, 13.33, 13.07, 12.22 and 12.22 respectively in 15 days after inoculation. The infection process was observed. The results indicate that the conidia germinate from the lateral sides in 12 hours after inoculation, and then form hyphae in 24 hours. The hypha extended and formed net-like mycelia in 36 hours and 72 hours, and finally produced conidiophores and chains of conidia in 96 hours.
  • Mycosystema. 2010, 29(5): 644-652.
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    The physiological characteristics of 32 main cultivated strains of Auricularia auricula-judae in China were analyzed, and the 27 indexes were evaluated and clustered by UPGMA and PCO. The results demonstrated that the cultivated germplasm of A. auricula-judae in China are richly endowed with genetic diversity, and the 32 strains analyzed are different physiologically. The strains can be divided into 3 groups, group of strains mainly cultivated in northeast region, group of strains in central and southeast regions, and group of strains in north, south and part of central regions. It is revealed that the physiological characteristics significantly correlated with geographical origins. The cultivated strains in the same region are more similar physiologically and some strains are suspected to be synonymous.
  • Mycosystema. 2010, 29(5): 653-664.
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    The reaction parameters and primers of ISSR-PCR for Ampelomyces quisqualis (Aq) were optimized by an orthogonal design and a single factor test. The genetic diversity of Aq was analyzed by the ISSR-PCR reaction system. Polymorphic bands of 73 isolates generated by five primers indicate that high diversity is conserved in Aq. The dendrogram based on ISSR results shows that the genetic diversity of Aq was relatively associated with the diversity of host fungi and host plants.
  • Mycosystema. 2010, 29(5): 665-669.
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    The uredinial germination process of wheat stripe rust fungus Puccinia striiformis f. sp. tritici on leaf surface of the host plant, wheat, and non-host plant, rice, and on the surface of wheat spike and stem was observed by fluorescence microscope and scanning electron microscope. The observation shows that after urediospores germinated and formed germ tube, a series of infection structures such as substomatal vesicle, infection hypha and haustorial mother cells were formed on the wheat leaf surface. The differentiation of haustorial mother cells could be found on the surface of glume, lemma, palea and stem of wheat. On rice leaf surface, the infection structure differentiation of the rust fungus could also occur. The observation by fluorescence staining revealed that no obvious difference was found of the differentiation of infection structures of the wheat stripe rust fungus between in vitro and in plant tissue.
  • Mycosystema. 2010, 29(5): 670-677.
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    Cordyceps militaris has been commercially cultivated. However, the degenerate subcultures of C. militaris have increasingly reduced the production of this economically important fungus. The characteristics of dehydrogenase activity, decolorization activity, mating-type and dsRNA infection of the degenerate strains were analyzed for comparison with the wild strains of C. militaris. The results showed that the degenerate strains had significantly lower dehydrogenase activity and decolorization activity, while no differences in characteristics of mating-type and dsRNA infection were observed between degenerated strains and normal strains. The result provides a quick method to detect early degeneration of C. militaris inocula during the process of artificial cultivation.
  • Mycosystema. 2010, 29(5): 678-682.
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    A fungal strain Curvularia lunata JQH-100 with high laccase-producing ability was isolated from maize leaves infected by curvularia disease. The highest laccase activity was found in the third day when Curvularia lunata JQH-100 was cultivated in the liquid medium. The optimum temperature and pH of crude enzyme using 2,2¢-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) as the substrate were 30℃ and 2.8, respectively. Several synthetic dyes were added into the culture medium to test the decolorization ability of Curvularia lunata. The results showed that the co-culture system decolorized 92.6% of Alizarin Red and more than 80% of Neutral and Congo Red. Laccase from Curvularia lunata was also purified for further assay of decolorizing Alizarin and Congo Red, which resulted in a decolorization percentage of 82.1% for Alizarin Red and 81.2% for Congo Red. This indicates that Curvularia lunata JQH-100 may be applied for treating dye-contaminated wastewater.
  • Mycosystema. 2010, 29(5): 683-690.
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    Using the combination of ethanol precipitation and DEAE-Sepharose Fast Flow chromatography, two forms of extracellular β-glucosidase from Aspergillus niger in bean dregs broth, designated as BGL1 and BGL2, were purified with a recovery of 25% and 23%, respectively. By running SDS-PAGE, their molecular weights were determined to be 102kDa for BGL1 and 97kDa for BGL2. At pH 2.5 and 55℃, BGL activity for hydrolyzing geniposide reached maximum, and was activated by Na+ , but inhibited to varying degrees by Mg2+, Ba2+, Cu2+, Fe2+, Zn2+and Hg+ at 10mmol/L. The inhibition of metal ions on BGL1 were stronger than that on BGL2. For all this, no significant effect of K+ on BGLs was observed. Ca2+ activated BGL1 significantly but had no impact on BGL2, while Fe3+ inhibited BGL1 but activated BGL2. Though p-nitrophenyl-d-glucopyranoside (pNPGlu) was the best substrate for BGL1 and BGL2 hydrolyzing pNPGlu, p-nitrophenyl-d-galactopyranoside, salicin and geniposide, the hydrolytic efficiencies of BGLs to geniposide were the highest. The Km values of BGLs to pNPGlu were 0.764mmol/L and 1.934mmol/L, and the Kcat/Km values of BGLs to geniposide were 4.28×104 and 1.04×105L/mol·s, respectively. BGL1 was highly stable over pH range of 2.0-7.0 at 20℃, and exhibited a half-life of 10min at 55℃, while BGL2 showed a wider range of pH stability (from 2.0 to 8.5) and the activity retained approximately 60% of the original activity after incubation for 60min at 55℃.
  • Mycosystema. 2010, 29(5): 691-697.
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    A chitinase gene was isolated from Thermoascus aurantiacus var. aurantiacus through RT-PCR and Tail-PCR methods. The full-length cDNA of 1,253bp contains an ORF of 1,197bp encoding 398 amino acids. Analysis of the deduced amino acid from nucleotide sequence revealed high homology with the catalytic domains of the glycoside hydrolase family 18. The chitinase gene was expressed in Pichia pastoris GS115. After induction for six days using methanol the transformed strain reached the highest production level and the expression amount was 0.433g protein per litter. The activity of expressed protein was 28.96U/mg. The molecular mass of a single band of the enzyme was estimated to be 43.9kDa by SDS-PAGE analysis. The optimum temperature of the enzyme was 60℃ and it maximal activity was at pH 8.0. The enzyme was thermostable at 50℃ and 60℃ and could still keep 45% activity at 70℃ after 30min, thus showing a potential for commercial uses.
  • Mycosystema. 2010, 29(5): 698-706.
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    A pathogen is usually enhanced its infectivity through activating and repressing protein kinase signal pathway in some way or other. Protein kinases play important roles in signal identification and transduction at the preliminary stage of plant-pathogen interactions. It was showed that there were abundant simple sequence repeats (SSRs) in the protein kinase genes of Magnaporthe grisea. The SSRs were analyzed within the extron, intron, 5 untranslated region (5′-UTR) and 3 untranslated region (3′-UTR) of 89 protein kinase genes, respectively. The effect of SSR expansion and contraction on protein secondary structure was predicted in gene coding regions. As compared with other regions, tri-nucleotide and hexa-nucleotide were more frequently observed in coding region and G+C content was more abundant in the SSR motifs. In addition, the frequency of hydrophilic amino acids was relatively higher. Depending on the size variation among field isolates of the pathogen, the expansion or contraction of amino acids encoded by SSR sequences potentially affected on secondary structure of protein. The data of this study indicated that the mutation of SSR might play an important role for pathogenic gene mutation.
  • Mycosystema. 2010, 29(5): 707-712.
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    mRNA differential display reverse transcription-PCR (DDRT-PCR) analysis with 24 pairs of primers was used to analyze mRNA differences between the wild type of Agaricus bisporus strain CGMCC No. 0214 and its substrate-decomposing ability degeneration strains 0214-3 and 0214-5 cultured in 4 kinds of different liquid media. As a result, 8 pairs of primers showed repeatable differences, and 9 special fragments were cloned. Sequencing analysis showed that 9 special fragments represented 5 different genes. These genes have high homologies with an ATP-binding cassette transporter subfamily A protein, carbohydrate esterase family 4 protein, chitin deacetylase, acetyl xylan esterase II and some other hypothetical proteins of several kinds of organisms. The results of this study provided a basis for further study of the molecular mechanism of substrate degradation ability degeneration of A. bisporus.
  • Mycosystema. 2010, 29(5): 713-718.
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    Four selenium polysaccharides, SeAPⅠ-1, SeAPⅠ-2, SeAPⅡand SeAPⅢ were isolated from Auricularia auricula-judae grown in selenium medium by DEAE52-cellulose and Sephacryl S-500 HR chromatography. The four polysaccharides had no absorption at 280 or 260 nm in their UV spectrum, indicating the absence of protein and nucleic acid. The high performance gel permeation chromatography (HPGPC) profile showed single and symmetrically sharp peaks, indicating that they were homogeneous polysaccharides. Effects of the four polysaccharides on the free fatty acid (FFA) and lipid mobility of S180 cell membrane were investigated. The results demonstrated that all the four selenium polysaccharides have good antitumor activity, and the activities of SeAPⅠ-2 and SeAPⅡ are significantly stronger than those of SeAPⅠ-1 and SeAPⅢ (p<0.05).
  • Mycosystema. 2010, 29(5): 719-725.
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    The enrichment of heavy metals in Antarctic environments has increasingly evoked public interests. Lichens have been widely used in study of heavy metal atmospheric deposition in many ecosystems. Although a few studies using lichens to monitor atmospheric heavy metal deposition have been conducted in Antarctica, differences in enrichment capability of atmospheric heavy metals were not known among Antarctic lichens. Five lichen species collected from Fildes Peninsula, King George Island, western Antarctica, viz. Caloplaca regalis, Himantormia lugubris, Ramalina terebrata, Sphaerophorus globosus and Usnea aurantiacoatra were used to analyze differences in enrichment capability of Co, Cr, Pb and Cu. After being exposed to air for a period of 2 months at an urban site of Baoding, Hebei Province, heavy metal contents in lichen thallus were measured by atom absorption spectrometry. The results show that all species can enrich atmospheric Pb, in which Himantormia lugubris shows the highest capability, followed by Usnea aurantiacoatra and Ramalina terebrata. Usnea aurantiacoatra and Himantormia lugubris can enrich atmospheric Cu at the similar level, followed by Sphaerophorus globosus, while the other two species show no enrichment of atmospheric Cu during the experiment. The significant enrichment of atmospheric Co and Cr was only observed in Himantormia lugubris and Usnea aurantiacoatra, respectively. Our results indicate that a combination of Usnea aurantiacoatra and Himantormia lugubris is an indicator with great potential for monitoring atmospheric deposition of Co, Cr, Cu and Pb, and Himantormia lugubris is also applicable to the monitoring of atmospheric deposition of Co, Cu and Pb.
  • Mycosystema. 2010, 29(5): 726-731.
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    Five new polyketides, chaetomones A-E (1-5), have been isolated from solid cultures of Chaetomium sp. by silica gel column chromatography (CC), Sephadex LH-20 CC and RP HPLC. The structures of these metabolites were determined mainly by analysis of their NMR spectroscopic data. Compound 5 displayed antifungal activity against Candida albicans (ATCC 10231) with an IC50 value of 20.0μmol/L.
  • Mycosystema. 2010, 29(5): 732-738.
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    A fungus strain F5 isolated from the rhizospheric soil of Bruguiera gymnorrhiza was identified as a member of Aspergillus section Circumdati. The morphological characters and internal transcribed spacer (ITS) sequence showed that this species is closely related to A. ochraceopetaliformis. This strain can produce antibacterial metabolites when cultured in modified glucose peptone yeast medium (GPYM) under the condition of 160r/min at 28℃ for 7 days in a rotary shaker. Two compounds were isolated from the fermentation broth and mycelial extract, and they were elucidated as (R)-(-)-mellein (Ⅰ) and flavacol (Ⅱ). Compound Ⅱ inhibited the growth of Bacillus subtilis with the minimum inhibitory concentration (MIC) value of 32mg/L, while the compound Ⅰ had no antibacterial activities.
  • Mycosystema. 2010, 29(5): 739-745.
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    The endophytic fungus strain ZD6 isolated from the stem of Bruguiera gymnorrhiza was identified as Penicillium citrinum by its morphological characters and internal transcribed spacer (ITS) sequence. This strain can secrete antibacterial metabolites when cultured in modified medium (consisted of 1% maltose, 2% mannitol, 1% sodium glutamate, 0.5% peptone, 0.15% yeast extract, 200g/L potato extract, pH6.5) under the condition of 160r/min at 28℃ for 7 days in a rotary shaker. Four compounds were isolated from the ethyl acetate and n-butanol extraction of its fermentation broth. These compounds were elucidated as emodin, cyclo-(Ala-Gly), erythritol and mannitol by MS, 1H-NMR, 13C-NMR and physiochemical parameters, among which emodin and erythritol were first reported from Penicillium citrinum. Emodin and erythritol showed stronger inhibition to the growth of Bacillus subtilis with minimum inhibitory concentration (MIC) value of 25μg/mL and 50μg/mL respectively, while emodin showed moderate inhibition to the growth of Pseudomonas aeruginosa with MIC value of 100μg/mL.
  • Mycosystema. 2010, 29(5): 746-752.
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    The decolorization ability of Curvularia lunata mycelial pellets, formed during the growth process was tested by using several synthetic dyes. The result showed that more than 80% of the tested dyes could be removed within 24h. The mycelial pellets demonstrated a good stability and could be used for 6 times. The decolorization percentage of malachite green was used as an index to optimize the production conditions for C. lunata mycelial pellets. The result of orthogonal experiment indicated that the optimal conditions were: glucose 20g/L, ammonium sulfate 5g/L, pH 5, potato 200g/L, KH2PO4 3g/L, MgSO4 5mg/L, CuSO4 0.5mg/L, VB1 5mg/L and rotation rate 120r/min. The optimized basal culture medium (without MgSO4, CuSO4) supplemented with microelements Cu2+, Mn2+, Mg2+ or Ca2+ had obvious promotion effect on decolorization of malachite green. In contrast, the medium added with Fe2+ caused a negative effect on decolorization. The mycelial pellets from the optimized basal culture medium supplemented with Zn2+, Al3+ or Na+ had a similar decolorization ability as compared with the contrast. Furthermore, the mycelial pellets prepared from optimized medium could efficiently decolorize wastewater containing dye mixtures.
  • Mycosystema. 2010, 29(5): 753-759.
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    Two endophytic fungal strains, Trichoderma longibrachiatum CSN-18 and Aspergillus sp. CSN-3, were isolated from healthy leaves of Camellia sinensis. The inhibitory activity against plant pathogenic fungi of these two fungi in the mixed culture was compared with monoculture. The results showed that the broth and its acetic ether extract of the mixed culture had much stronger inhibitions against Magnaporthe grisea, Phyllosticta camelliaecola, Guignardia camellia and other 5 species of plant pathogenic fungi as compared to monocultures of CSN-18 and CSN-3. The inhibition rates of acetic ether extract of the mixed culture to the mycelial growth of P. camelliaecola, M. grisea and G. camellia were 79.48%±1.46%, 76.99%±0.91% and 71.51%±4.93%, respectively. Inhibition rates of acetic ether extract of the mixed culture to the spore germination of P. camelliaecola, G. camellia and M. grisea were 100.00%, 90.90%±2.59% and 84.00%±5.29%, respectively. The broth of the mixed culture had a much higher inhibition rate to the mycelial growth of P. camelliaecola as compared to monocultures. Spore germination of P. camelliaecola was promoted by broth of the mixed culture in the concentration lower than 30%. But spore germination was dramatically inhibited when broth concentration was higher than 40%.
  • Mycosystema. 2010, 29(5): 760-766.
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    Aqueous ethanol was used as solvent to optimize the conditions for maximum extraction of polyphenols from sclerotia of Phaeoporus obliquus by orthogonal test method in which ethanol concentration, extraction time, temperature and the volume ratio of material and solvent were taken into account as factors. The results showed that the maximum extraction of polyphenols (3.099% m/m) was obtained by 50% aqueous ethanol with a ratio of material and solvent at 1:10, extraction temperature at 50℃ and extraction time at 45min. Aqueous ethanol is a safe, economical and effective solvent for extracting polyphenols from Paeoporus obliquus.
  • short communications
  • Mycosystema. 2010, 29(5): 767-770.
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  • Mycosystema. 2010, 29(5): 771-776.
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  • Mycosystema. 2010, 29(5): 777-781.
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