15 July 2010, Volume 29 Issue 4

    

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Papers
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  A new species of Phomopsis on Castanea mollissima

  Mycosystema. 2010, 29(4): 467-471.

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  A new leaf disease was found on chestnut in Tai’an and Linyi of Shandong Province. The infected leaves firstly produced small spots and then turned brown and finally fell off. Pathogenic separation, purification, morphological observation and rDNA-ITS sequence analysis of the pathogen indicated it was a new species, Phomopsis castaneae-mollissimae. Brief description of the species was provided. The holotype specimen is deposited in the herbarium of Shandong Agricultural University: Plant Pathology (HSAUP).
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  Two new species of Humicola from Guizhou Province, China

  Mycosystema. 2010, 29(4): 472-474.

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  Two new species of Humicola, Humicola guizhouensis and H. terrestris isolated from soil of Guizhou Province of China are described and illustrated. The dried specimens (holotypes) and living cultures examined have been deposited in the Herbarium of Guizhou University: Plant Pathology (HGUP).
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  The causal agent of coelomycete leaf spot of Paeonia suffruticosa and its synanamorphs

  Mycosystema. 2010, 29(4): 475-480.

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  A new Paeonia suffruticosa leaf spot disease firstly reported by Zhang et al. from Henan Province was also observed on the same plants growing in Heze, Jinan, Tai’an and Feicheng etc. of Shandong Province in 2007－2008. The disease can also infect stems and branches of the plant to form grey-whitish and brown-margined spots on which a few black and flattened fructifications scattered. The conidiomata on leaf spots are cupulate whereas those on stem spots are semi-spherical and unostiolate. However, the morphology of the conidiophores, conidiogeneous cells and conidia in these different types of conidiomata are very similar. The 7 isolates from stem spots and 21 ones from leaf spots are also similar in cultural and biological characteristics as well as pathogenicity. Based on morphological observation and rDNA-ITS sequence analysis, the pathogen on leaf spots was identified as Hainesia lythri, while that on stem spots as Pilidium concavum. They are synanamorphs of the same fungus Discohainesia oenotherae.
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  Identification and characterization of the fungal strain MT-06 for biocontrol of strawberry anthracnose

  Mycosystema. 2010, 29(4): 481-487.

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  A study was conducted to determine the biological characteristics of the fungal strain MT-06 isolated from the strawberry tissue suffered from anthracnose disease, which was identified as Talaromyces flavus (Kl?cker) Stolk & Samson based on morphologic observation and both rDNA-ITS and β-tubulin sequence. The results indicate that the colony of MT-06 grows better on the PDA and potato saccharose agar (PSA) media, while Czapek medium was most favorable to its conidial production. The optimal temperature for its colony growth and sporulation ranged from 28 to 35℃ and from 25 to 28℃, respectively. The optimal initial pH of culture media was pH5. The strain could grow in either dark or light, but lighting was beneficial for vegetative growth and sporulation in comparison with dark. The test of antimicrobial spectrum shows that MT-06 is able to inhibit many kinds of plant fungal pathogens in vitro.
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  Morphological and molecular identification of citrus anthracnose pathogen from Guangdong Province

  Mycosystema. 2010, 29(4): 488-493.

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  Anthracnose is one of the major fungal diseases of citrus. A large-scale outbreak of anthracnose in Citrus sinensis cv. gonggan, the high-yield and high-quality cultivar of citrus, occurred in Deqing county, Guangdong Province in the spring of 2007. For identifying the pathogen so as to control this disease effectively, samples of citrus anthracnose were collected from ten cultivars of the genus Citrus in six areas of Guangdong Province, and a total of 75 single-conidial isolates were obtained, among them 10 representative single-conidial isolates were identified at the species level. Based on cultural and morphological characteristics, ribosomal DNA (rDNA) internal transcribed spacer (ITS) sequence analysis, ITS-specific primers PCR detection and phylogenetic relationships with other species of Colletotrichum, the 10 isolates were identified as Colletotrichum gloeosporioides, whereas C. acutatum, the causal agent of postbloom fruit drop of citrus seriously occurring in some countries in the world, was not yet found in this research.
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  Pythium heterothallicum new to China and its pathogenicity

  Mycosystema. 2010, 29(4): 494-501.

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  Pythium species were isolated by using petal baiting and tissue isolating methods from rhizosphere soils of bean, grape, cabbage and corn in Lanzhou and Zhangye, Gansu Province during September and October, 2007. The Pythium colonies were taken at the colony edge for subculture and purification. The purified isolates were primarily identified based on morphology and colony characteristics. It was found that seven Pythium isolates were similar in morphology and colony. Analysis of rDNA-ITS sequence and β-tubulin gene sequence of the representative isolate P3-1A indicated that the isolates were Pythium heterothallicum, a newly recorded species in China. P. heterothallicum is heterothallic. The pathogenicity of the seven isolates were tested on corn, tomato and cucumber by using soil burying method. The results indicate that the inoculated corn showed rot root, brown and necrotic root tips, rot basal stems, growth suppression and stunting, but no obvious symptoms on tomato and cucumber. It is concluded that P. heterothallicum was pathogenic to corn but not to tomato and cucumber.
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  Isolation and identification of an endophytic fungus producing resveratrol in Polygonum cuspidatum

  Mycosystema. 2010, 29(4): 502-507.

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  The fresh roots of Polygonum cuspidatum, a traditional Chinese medicine plant, were used to isolate endophytic fungi. Fifteen strains were obtained. Five isolates producing resveratrol were investigated based on HPLC detection after fermentation in potato dextrose broth (PDB). Strain H8 was the most prospective in biological production of resveratrol. Morphological characteristics and internal transcribed spacer (ITS) sequences show that strain H8 was Fusarium verticillioides.
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  Population structure of Phytophthora infestans on tomato in Hebei Province

  Mycosystema. 2010, 29(4): 508-517.

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  Tomato late blight is one of the most devastating diseases in tomato production in Hebei Province. The study of the population structure of Phytophthora infestans on tomato is contributive to the control of the disease. The phenotypes of 49 isolates collected in Baoding, Cangzhou and Tangshan cities of Hebei Province during 2007 and 2008 were investigated for mating types and metalaxyl resistance respectively by means of dual culture with A1 and A2 testers on rye plates and radical growth assay on fungicide-amended media in vitro. The results show that all the tested isolates were A1 mating type, their majorities were metalaxyl-sensitive and only seven isolates were metalaxyl-resistant. The genotypes were also assessed for mitochondrial DNA haplotypes with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), simple sequence repeats (SSR) and amplified fragment length polymorphism (AFLP). Only one DNA haplotype (Ia) and three SSR genotypes (type I, type II and type III) were determined in this study. However, three AFLP clades (α, β and γ) were formed at a genetic similarity value of 0.87 and all isolates of tomato Phytophthora infestans from Hebei were included in α clade, which were further divided into seven sub-clades. The seven AFLP sub-clades were not associated with metalaxyl resistance and geographical origin, however, type II SSR genotype was correlative to metalaxyl resistance and geographical origin. The phenotypic and genotypic data indicate that the population structure of Phytophthora infestans from tomato in Hebei Province was lower in genetic diversity.
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  Mycobiotal investigation of natural Ophiocordyceps sinensis based on culture-dependent investigation

  Mycosystema. 2010, 29(4): 518-527.

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  Natural Ophiocordyceps sinensis, a traditional Chinese medicine, is endemic to alpine regions on the Tibetan Plateau. Various fungal species have been reported to be associated with natural O. sinensis. In this study, the mycobiota of natural O. sinensis was investigated with the traditional culture-dependent method. In total, 572 fungal cultures were isolated from different parts (stromata, sclerotia, and external mycelial cortices) of natural O. sinensis. Most of these cultures were assigned to 37 genera according to morphological characteristics, and 92 OTUs were identified at the 97% similarity threshold according to their nrDNA ITS sequences following SSCP analyses. The number of isolates and OTUs of Ascomycetes was larger than that of Basidiomycetes and Zygomycetes. More isolates and OTUs were obtained from cortices than from sclerotia and stromata. The most abundant species from stromata was Penicillium chrysogenum, while that from both sclerotia and cortices was Pseudogymnoascus roseus. Several unclassifiable fungal species were also detected in this study, which could be novel species.
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  Laccase transformation system established in Hypsizigus marmoreus breeding

  Mycosystema. 2010, 29(4): 528-535.

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  The laccase transformation system in Hypsizigus marmoreus breeding was established by enzymological technique in combination with protoplast fusion. Protoplast of Hypsizigus marmoreus and heat-treated protoplast of Pleurotus sajor-caju with faster growth and higher activity of laccase were fused by fusogen. Two fusion strains III18C, III2A were screened by the RB-PDA plate. Antagonistic test, RAPD-PCR and amplification of laccase gene were used to study the genetic stability of fusion strains and two parents. The results showed that there was an obvious antagonistic line between fusion strain and two parents. The fusion strain’s amplification patterns of RAPD-PCR were different from parents, and a partial gene of laccase was amplified. Using laccase transformation system to screen fusion strains has shown an effective advantage.
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  Purification and characterization of a xylanase from psychrotrophic Eupenicillium crustaceum

  Mycosystema. 2010, 29(4): 536-541.

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  A xylanase from psychrotrophic Eupenicillium crustaceum was purified and characterized. The xylanase was purified to homogeneity by ammonium precipitation and anion-exchange chromatography and its subunit molecular mass was estimated to be 35kDa by SDS-PAGE. The xylanase showed apparent optimal activity at pH 5.5 and 50℃. It retained more than 80% of its maximum activity at pH 4.5－6.5 and retainded 40% of its maximum activity at 20℃. The enzyme was greatly activated by Ag+ and Fe2+, and strongly inhibited by Mn2+ and Hg2+. The xylanase displayed a strict substrate specificity.
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  Expression analysis of Alternaria solani Hog1 MAPK homologous gene AsHog1 and correlation between the gene and fungicidal resistance of the pathogenic fungus

  Mycosystema. 2010, 29(4): 542-547.

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  Degenerate PCR primers were designed according to conserved amino acid sequences of several filamentous fungus MAPKs (mitogen-activated protein kinase), and a partial DNA fragment encoding a MAPK of Alternaria solani (designated AsHog1) was cloned using PCR. The expression profiles of AsHog1 between iprodione-sensitive and -resistant strains under conditions of osmotic pressure and fungicide stress were compared using real-time RT-PCR. When the sensitive isolate QX21 was exposed to 0.8mol/L NaCl or 2mg/L iprodione, the expression level of AsHog1 increased and reached the maximum at 12h after induction, which was up to 4.12 and 25.23-fold higher than those of their controls, respectively. After 16h, the expression levels were slightly decreased. However, no obvious change in gene expression patterns was observed in resistant isolates either induced in the laboratory or naturally occurred in the field after exposure to the osmotic pressure or fungicide stress. The results indicated that AsHog1 was probably correlated with iprodione resistance in A. solani. This study is helpful to further study the mechanism of iprodione resistance in A. solani, to explore molecule-based techniques to detect iprodione resistance, and to develope strategy for the resistance management.
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  Ginsenoside Re biotransformed by Rhizopus sp.

  Mycosystema. 2010, 29(4): 548-554.

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  Ginsenoside Re was transformed by Rhizopus sp., and Re fermentation products were compared by HPLC system analysis. Liquid chromatography (LC) and mass spectrum (MS) analysis showed the conversion ratio of ginsenoside Re was 92.16%. The raised compound 1 in the panaxatriol fermentation products was identified as ginsenoside Rg2 with yield of 10.13%, while compound 2 and compound 3 respectively as ginsenoside Rg3 and Rk1 with yield of 29.23%. The results showed that Re can be biotransformated by Rhizopus sp. It was inferred that ginsenoside Re had been biotransformed into ginsenoside Rg2. The transformation of ginsenoside Re into Rg5/Rk1 needs further studies.
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  The effects of stressed conditions on the accumulation of phenolic compounds by Penicillium sp. No. T4 and their subsequent antioxidant activities

  Mycosystema. 2010, 29(4): 555-560.

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  This study was designed to evaluate the effects of stress conditions on the accumulation of phenolic compounds by Penicillium sp. No. T4 in solid substrate and their subsequent antioxidant activities. H2O2, ultraviolet and medium of nutrition deficiency were used as oxidative stressors. Phenol levels were measured using Folin-Ciocalteu method. Capacities for scavenging free radicals were detected by spectrophotometric methods. The exposure of Penicillium sp. No. T4 to H2O2, at 1mmol/L or ultraviolet radiation for 40s and 50% nutrition deficiency substrate resulted in a higher production of phenolic compounds as compared with control, with levels reaching 63.86mg/g, 58.4mg/g and 43.19mg/g, respectively. Extracts from mycelia grown under stress conditions showed higher capacities for scavenging hydroxyl radical than other free radical, with a scavenging rate of 35.28% by the phenolic compounds from mycelia grown in 1mmol/L H2O2, 69.97% by those from mycelia under 40s ultraviolet radiation and 50.83% by those from mycelia under 75% medium nutrition deficiency. The results suggest that Penicillium sp. No. T4 accumulates more phenolic compounds under oxidative stresses and thereby enhances its resistance to oxidative stresses.
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  Induction of tomato signal substances by arbuscular mycorrhizal fungi

  Mycosystema. 2010, 29(4): 561-568.

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  In order to evaluate the induction of plant signal substances by arbuscular mycorrhizal (AM) fungi, a pot experiment by using Lycopersicon esculentum seedlings inoculated with Glomus mosseae, G. versiforme, G. etunicatum, G. intraradices and Gigaspora margarita was carried out. The content of endogenous salicylic acid (SA), jasmonic acid (JA), nitrogen monoxide (NO) and hydrogen peroxide (H2O2), the related resistant enzyme activity, and malondialdehyde (MDA) contents in tomato seedlings were determined in 35 days after inoculation. The results showed that AM fungal inoculation increased significantly the seedling growth (plant fresh weight, plant height, shoot and root dry weight), SA, JA, NO, H2O2 and SA glucosidate content in roots and leaves. The maximum induction effects occurred in G. mosseae and the content of NO, JA, H2O2 and SA glucosidate in leaves and roots had increased by 4.3 and 2.9 times, 7.8 and 9 times, 1.9 and 2.2 times, and 2.9 and 3.6 times respectively over the control. The free SA content in mycorrhizal plants was lower than that in control, but the content in plant treated with G. mosseae was a little higher than that in the control at induction peak. Superoxide dismutase (SOD), peroxidase (POD), phenylalanine ammonia-lyase (PAL) and peroxide catalase (CAT) activities in mycorrhizal plants were greater than those in control. G. mosseae gave the maximum induction effectiveness, and the activity of SOD, POD, CAT, and PAL in roots and leaves had increased by 1.6 and 1.3 times, 8.9 and 4.1 times, 1.4 and 2.2 times, 3.3 and 2.9 times as compared with the control; the G. etunicatum gave the minimum induction effectiveness, and the activity of SOD, POD, CAT and PAL in roots and leaves had increased to 0.26 and 0.14 times, 2.3 and 1.0 times, 0.1 and 0.28 times, 0.55 and 0.31 times as compared with the control; MDA content reduced by 66% and 68%, 34% and 41%, 51% and 50%, 12% and 26%, 18% and 29% as compared with the control. It was concluded that AM fungi could simultaneously induce many plant signal substances, and these signal substances were involved in the resistant expression in AM fungi-tomato symbiont.
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  Antioxidant activities of extracts and sub-fractions from Tuber indicum

  Mycosystema. 2010, 29(4): 569-575.

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  Tuber indicum were extracted under reflux of 55% ethanol (ECE), which were further extracted by petroleum ether and ethyl acetate, respectively. The antioxidant activities of ethanolic crude extract (ECE) and two solvent sub-fractions [namely, petroleum ether fraction (PEF) and ethyl acetate fraction (EAF)] were investigated using three different in vitro antioxidant assays, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, hydroxyl radical scavenging activity and chelating ability. The results showed that ECE and two sub-fractions possessed significant difference (P＜0.05) in capacities for scavenging DPPH, hydroxyl radical and chelating ferrous ion. In comparison, ECE showed the most potent capacity for scavenging DPPH with EC50 values reaching 1.61g/L. EAF exhibited the highest capacity for scavenging hydroxyl radical and ferrous ion chelating activities with EC50 values reaching 3.31g/L and 0.70g/L, respectively. TP content in EAF was 2.964mg GAE/g extract, and that in ECE 2.618mg GAE/g extract. The capacity for scavenging free radicals and ferrous ion chelating activity of PEF was the lowest among the three extracts, with a TP level reaching 1.124mg GAE/g extract. Thus antioxidant activity of Tuber indicum seems to be positively correlated with the content of total phenols.
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  Chemical structure of polysaccharide AAPS-3 from Auricularia auricula-judae

  Mycosystema. 2010, 29(4): 576-581.

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  A homogeneous polysaccharide AAPS-3 was obtained from Auricularia auricula-judae by countercurrent chromatography technology. The structure of AAPS-3 was elucidated using the method of complete acid hydrolysis, methylation, and 1D and 2D nuclear magnetic resonance (NMR). AAPS-3 was found to be mainly composed of Glc by means of total acid hydrolysis. Its molecular weight was estimated to be 4.83×105 by HPLC method. Methylation analysis and 1H and 13C NMR indicated the main chain of AAPS-3 consisted of β-D-1,4-Glc, β-D-1,3-Glc and β-D-1,6-Glc.
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  Isoprenylated chromones from the plant endophytic fungus Pestalotiopsis fici

  Mycosystema. 2010, 29(4): 582-587.

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  Pestaloficiols M－P (1－4), new isoprenylated chromone derivatives, have been isolated from a scale-up fermentation extract of the plant endophytic fungus Pestalotiopsis fici. The structures of these compounds were elucidated primarily by MS and NMR techniques. Compound 2 displayed inhibitory effects on HIV-1 replication in C8166 cells, whereas compounds 3 and 4 showed cytotoxic activity against the human tumor cell line HeLa. Compound 3 also showed antifungal activity against Aspergillus fumigatus.
short communications
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  Two species of Tubaria (Agaricales) new to China

  Mycosystema. 2010, 29(4): 588-591.

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  Notes on hyphomycetes of conifer needle I. Two newly recorded genera and three newly recorded species from China

  Mycosystema. 2010, 29(4): 592-594.

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  Three new Chinese records of Conidiobolus

  Mycosystema. 2010, 29(4): 595-599.

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  Plasmalemma structure and chromatin feature of Fuligo septica

  Mycosystema. 2010, 29(4): 600-602.

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  Assessment for protein nutrition of fruit bodies, mycelia and fermentation broth of Lyophyllum decastes

  Mycosystema. 2010, 29(4): 603-607.

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  Total phenolic content and antioxidant activities of the extracellular melanin from Lachnum sp. YM-223

  Mycosystema. 2010, 29(4): 608-611.

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  Acute toxicity and in vivo inhibiting effects upon Hepatoma H22 of ethanol precipitate of Antrodia camphorata culture filtrate

  Mycosystema. 2010, 29(4): 612-615.

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  Chemical composition in cultural fruit bodies of Gymnopilus spectabilis

  Mycosystema. 2010, 29(4): 616-618.

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  Chemical constituents and antibacterial activity of petroleum ether extract from fruit bodies of Tricholoma mongolicum

  Mycosystema. 2010, 29(4): 619-624.

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