15 January 2012, Volume 31 Issue 1

    

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Papers
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  A preliminary report of dematiaceous hyphomycetes from soil in Liaoning Province, China

  Mycosystema. 2012, 31(1): 1-7.

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  Dematiaceous hyphomycetes from soil in the different ecosystems of Liaoning Province were investigated. A total of 45 isolates belonging to 26 species in 18 genera were obtained from 28 soil samples. Among them, Bipolaris alocasiae and Humicola longistipes are new species. Specimens (dried cultures) and living cultures of all isolates studied have been deposited in the Herbarium of Shandong Agricultural University: Plant Pathology (HSAUP). Isotypes are kept in HMAS.
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  Dematiaceous hyphomycetes from soil in Songliao Plain and western prairie wetlands, Jilin Province, China

  Mycosystema. 2012, 31(1): 8-16.

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  Dematiaceous hyphomycetes from soil of different ecosystems in the areas of Songliao Plain and western prairie wetlands, Jilin Province, China were investigated. A total of 116 isolates were obtained from 42 soil samples. Fourty-six species in 25 genera were identified. Among them, Bipolaris obovoidea is a new spieces, while Monodictys bicolorata, Periconia hispidula and Torula herbarum f. quaternella are new records for China. Specimens (dried cultures) and living cultures of all isolates studied have been deposited in the Herbarium of Shandong Agricultural University: Plant Pathology (HSAUP). Isotype (dried cultures) is kept in HMAS.
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  A preliminary report of dematiaceous hyphomycetes from soil in tropical Yunnan, China

  Mycosystema. 2012, 31(1): 17-23.

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  A total of 53 isolates of dematiaceous hyphomycetes belonging to 37 species in 29 genera were isolated and identified from 30 soil samples collected in the tropical rain forest, botanical garden, wastelands and cultivated lands in Xishuangbanna and Hekou areas, Yunnan Province, China. Bloxamia nilagirica, Cephaliophora irregularis, Haplographium delicatum and Periconia britannica were newly recorded for China. All the specimens (dried cultures) and living cultures studied have been deposited in the Herbarium of Shandong Agricultural University: Plant Pathology (HSAUP).
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  Interception of Phytophthora syringae on Citrus fruits imported from California, USA

  Mycosystema. 2012, 31(1): 24-30.

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  Several brown rot fruits have been found from Californian fresh orange sample. Three isolates similar to Phytophthora syringae were obtained from these rot fruits. The colonies grew slowly on PDA and V8 juice agar (V8A). They were stellate, tiled and thin on V8A, but on PDA, the margin was chrysanthemum petaloid; mycelia dense, milk-white-colored. Sporangia and catenulate hyphal swellings formed in sterilizing water and soil extract water in 48h. The isolates were homothallic. Oospores were produced abundantly on PDA and V8A mixed with fresh Citrus fruit tissue or leaf of rhododendron. The isolates were wounded-inoculated on fresh Citrus fruits. Typical brown rot symptoms are apparent on inoculated fruits in 7d. DNA of hyphae was amplified with ITS1/ITS4 primers. In comparision with P. syringae from NCBI GenBank databases, the sequences had 99% identity. All the results of tests showed that the isolates were Phytophthora syringae.
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  Anastomosis grouping and genetic diversity of maize pathogen Rhizoctonia solani in Shandong

  Mycosystema. 2012, 31(1): 31-39.

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  One hundred and three isolates were obtained from the sheath blight samples of maize from 14 counties in Shandong Province. The cell nuclei were stained and observed under the fluorescent microscope. To identify the anastomosis group, the examined isolates were tested by pairing growth. The result showed that the isolates were assigned to the multinucleate groups AG-1-IA, AG-1-IB, AG-1-IC, AG-3, AG-4-HG-I, AG-5 and WAG-Z and binucleate group AG-Ba. Among the anastomosis groups, the type AG-1-IA was dominant in the population with a rate of 60.19%. Genetic diversity of the 103 isolates was investigated by the inter-simple sequence repeats (ISSR) analysis using 3 specific and stable primers. Forty-five sites were generated in total, among which 91.1% were polymorphic. The result indicated there was high genetic diversity among the populations. By UPGMA cluster analysis, the isolates were classified into six subgroups and the clustering had relationships in association with geographical origin and their anastomosis groups.
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  Population structure of Alternaria solani on potato in Hebei Province

  Mycosystema. 2012, 31(1): 40-49.

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  The population genetic structure of 145 isolates of Alternaria solani collected from potato in Hebei Province and 30 ones in other four provinces in China from 2009 to 2010 was revealed by phenotypes such as virulence of Mancozeb and sporulation capacity in combination with AFLP genotype. The test of virulences showed that all tested isolates of A. solani were sensitive to Mancozeb, with EC50 values of 1.04－4.86μg/mL, averaging 2.93μg/mL. Average sporulation quantity of the tested 47 isolates in Hebei Province and 30 ones in other four provinces used as control were 85 and 50 conidia/mm2 repectively, showing significant difference. AFLP genotypes revealed a high diversity of A. solani collected from potato and each isolate had a specific AFLP genotype. AFLP genotype of isolates was closely related to the geographic origin, and related to a certain extent to sporulation quantity, but not correlated with sensitivity of mancozeb to pathogen.
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  Hymenochaete in China 1. Three species new to China from Guangdong and Anhui provinces

  Mycosystema. 2012, 31(1): 50-54.

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  Three wood-decaying fungi in the genus Hymenochaete, H. berteroi, H. tasmanica and H. tenuis are newly recorded in China. H. berteroi and H. tenuis were collected from Guangdong Province, while H. tasmanica was collected from Anhui Province. H. berteroi is characterized by its submonilioid hyphae, presence of setal hyphae and relatively small basidiospores. H. tenuis is unique for its thin basidiocarps and oblong-ellipsoid or short-cylindrical basidiospores in section Gymenochaete. H. tasmanica is distinguished by its stratified context and presence of dendrohyphidia in the hymenium. Illustrated descriptions of these three species are provided based on the Chinese materials.
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  New records of Galerina collected from Changbai Mts. in China

  Mycosystema. 2012, 31(1): 55-61.

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  Four species of Galerina collected from Changbai Mountains in Jilin Province, G. oregonensis, G. pistillicystis, G. stagnina and G. uncialis, were reported for the first time in China. The morphological descriptions and illustrations were given in this paper. The examined specimens are deposited in the Herbarium of Mycology of Jilin Agricultural University (HMJAU).
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  Four new Chinese records of arbuscular mycorrhizal fungi

  Mycosystema. 2012, 31(1): 62-67.

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  Four new Chinese records Glomus arenarium, G. mortonii, G. trimurales and Funneliformis xanthium were isolated from the root zone soils of Prunus mume and coastal plants. They were redescribed and illustrated herein based on the Chinese materials. These specimens were stored in the Herbarium Mycologicum Academiae Sinicae (HMAS) and Xiamen Botanical Garden Herbarium.
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  Biological characteristics of tomato powdery mildew Oidium neolycopersici

  Mycosystema. 2012, 31(1): 68-73.

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  Biological characteristics of Oidium neolycopersici, the main agent causing tomato powdery mildew, were investigated. The test revealed that the conidia germinated at the optimal temperature of 20－25℃ and the optimal relative humidity of 80%－100%, but failed to germinate in water drop. It was also found that the conidial germination had no strict requirement for illumination and pH value. They could germinate at pH3－12 with above 90% germination percentage. The fungus could utilize various carbon sources, and mannitol and galactose are the best of all. Among nitrogen sources, NO3-N (nitrate) is the best, but NH4-N and organic nitrogen could inhibit the conidial germination. The lethal temperature for conidial germination was 44℃ for 10min.
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  The hyperparasite Ampelomyces quisqualis and its host diversity in Laoshan Mountain of Qingdao

  Mycosystema. 2012, 31(1): 74-85.

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  One hundred and forty-three powdery mildew fungus samples collected from 35 species of plants belonging to 30 genera of 16 families in Laoshan Mountain of Qingdao during 2005 and 2007 were morphologically identified as 22 species of 7 genera. Of which 40 samples were found to be colonized by Ampelomyces quisqualis. Further investigations indicate that its host plants include 15 species belonging to 14 genera of 8 families, and its host powdery mildew fungi include 10 species belonging to 7 genera. The statistical analyses showed that A. quisqualis had the highest incidence and intensity of mycoparasitism on Arthrocladiella mougeotii. The analyses of community composition implied that Sphaerotheca fusca was the dominant host fungus of A. quisqualis with the highest importance value of 89.22%. Asteraceae, with the highest importance value of 97.85%, was the dominant host plant family.
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  Se-enriched characteristics of the mushroom Flammulina velutipes

  Mycosystema. 2012, 31(1): 86-91.

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  The characteristics of selenium (Se) enrichment and resistance in the mushroom Flammulina velutipes was studied. Se concentration was determined by ICP-MS. Se distribution in mushroom was investigated by using the selective and sensitive HPLC-ICP-MS. The results showed that Se concentration in samples was increased along with the increase of Se in the medium. The distribution of Se in the fruiting body in the same medium was as follows: pileolus cap>stripe>root. When the concentration of Se in mediums was below 30mg/kg, the growth of the Flammulitna velutipes was accelerated and the bioaccumulation coefficient of Se was between 2.8 and 9.9. The growth of mushroom was inhibited by the presence of 40－200mg/kg Se in media. It was not suitable for mushroom to grow when the concentration of Se >200mg/kg. The molecular weight of the detectable soluble compounds were under 25kDa after separation by SEC-HPLC. The SEC-HPLC-ICP-MS analysis indicated that the content of Se was 68.74－2,986.00μg/kg, accounting for 71.87% of the total amount and 4.88% of Se in Se-containing proteins. It was not only cost-effective, but also safe and non-poisonous to cultivate F. velutipes as a Se-carrier.
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  Analysis of volatile components of three entomogenous fungi by SDE-GC-MS

  Mycosystema. 2012, 31(1): 92-101.

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  Volatile components of Paecilomyces cicadae, Isaria gracilioides and Hirsutella heteropoda were extracted by method of simultaneous distillation extraction (SDE), and analyzed with gas chromatography-mass spectrometry (GC-MS). There were 44, 28 and 19 compounds were identified from the three species respectively. The main components were terpenoids, aromatic compounds, alcohols, alkanes, esters and aldehydes. Components’ comparison revealed that butylated hydroxytoluene, 1-octen-3-ol, benzeneacetaldehyde and decamethyl-cyclopentasiloxane existed commonly in all the 3 species. Besides the 3 main common components Paecilomyces cicadae contained uniquely 5-methylfurfuryl acetate, (E,E)-2,4-decadienal and decahydro-4,8,8-trimethyl-9-methylene-1,4-methanoazulene; Isaria gracilioides contained uniquely 5,6-dihydro-6-pentyl-2H-pyran-2-one, 2,4-dimethyl-oxazole, phenol and 1-ethenyl-1- methyl-2,4-bis(1-methylethenyl)-cyclohexane, and Hirsutella heteropoda contained uniquely 3,4,5-trimethoxy-Silane, (4-methoxyphenoxy)trimethyl-benzaldehyde, 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone, and 9-octadecenoic acid(Z)-2-hydroxy-1-(hydroxymethyl)ethyl ester.
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  The protoplast preparation and regeneration of Trametes hirsuta and laccase production of monokaryons

  Mycosystema. 2012, 31(1): 102-109.

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  Trametes hirsuta is very effective on degradation of lignin, and is proved to be of high value in biofuel, pulping, feed and other industries. To get the monokaryons of T. hirsuta YJ-9-1, we investigated the effect of several factors on the protoplast preparation and regeneration, such as growth medium type, fungal age, hydrolysis time, centrifugal speed for purification of protoplasts and regeneration medium type. The monokaryons were identified by using DAPI staining and clamp connection deficiency method, and ligninolytic enzyme production characteristics of the monokaryons were investigated. The protoplast forming number and regeneration rate reached 5.0×106/mL and 9.1% respectively under the optimal conditions, viz. yeast glucose malt agar (YGM) medium for fungus growth, fungal age of 88h, hydrolisis duration of 1h, centrifugal speed of 4,000r/min, and yeast glucose malt sucrose agar (YGMS) medium for protoplasts regeneration. Three strains of monokaryons were obtained and the strain named D-2-1 had a higher laccase production level than that of YJ-9-1, and the laccase activity of D-2-1 reached 771.67U/L on day 12, which is 1.5 times as high as that of YJ-9-1.
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  Purification and comparison of two lectins from the genus Russula

  Mycosystema. 2012, 31(1): 110-118.

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  Two novel lectins RDL and RLL were purified from fruiting bodies of Russula delica and Russula lepida using ion exchange chromatography and gel filtration. Based on the results of gel filtration and SDS-PAGE, both RDL and RLL were dimeric proteins with a molecular mass of 60kDa and 32kDa, respectively. The N-terminal amino acid sequences of RDL and RLL were determined to be GLKLAKQFAL and VWYIVAIKTDVPRTT, respectively. RDL was stable in temperature range of 20－70℃, HCl solution up to 25mmol/L, or NaOH solution up to 12.5mmol/L, but was inhibited by o-Nitrophenyl-β-D-galactopyranoside (25mmol/L) and inulin (50mmol/L). RDL manifested antiproliferative activity toward tumor cell lines hepatoma Hep G2 and mastocarinoma MCF7, and inhibitory activity toward HIV-1 reverse transcriptase (RT), with IC50 values of 0.88μmol/L, 0.52μmol/L, and 0.26μmol/L, respectively. RLL was stable in temperature range of 20－70℃, HCl and NaOH solutions up to 12.5mmol/L, but was inhibited by inulin (25mmol/L) and o-Nitrophenyl- β-D-galactopyranoside (100mmol/L). It showed antiproliferative activity toward Hep G2 and MCF7, with IC50 values of 1.60μmol/L and 0.90μmol/L, respectively, but was lacking in HIV-1 RT inhibitory activity.
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  The detection and analysis of cadmium in Lentinula edodes

  Mycosystema. 2012, 31(1): 119-126.

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  As a toxical element, cadmium in Lentinula edodes has been drawn much attention. In order to investigate the accumulation mechanisms of cadmium in Lentinula edodes, the effects of varieties, cultivation regions, times of harvest and morphological portions on accumulation of cadmium in Lentinula edodes have been studied. The results showed that, the distinction of cadmium content would be resulted from all the four factors above. The contents of cadmium in different part of Lentinula edodes fruit body were ranked as follow: gill > cap > stipe.
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  Isolation and identification of secondary metabolites from the solid culture of Flammulina velutipes

  Mycosystema. 2012, 31(1): 127-132.

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  The ethyl acetate extracts of the solid culture of Flammulina velutipes was subjected to silica gel, ODS, sephadex LH-20 column chromatography and HPLC prepration. Four sesquiterpenoids have been isolated from the mycelial culture of this mushroom. One novel eudesmane-type sesquiterpenoid together with three known cuparene-type sesquiterpenoids were obtained, including flamvelutpenol A (1), aquaticol (2), enokipodin C (3), and limacellone (4). The absolute configuration of 1 and 4 were determined by forming complex with [Rh2(OCOCF3)4] and then applying the bulkiness rule. Compounds 3 showed strong antibacterial activity against methicillin-resistant Staphylococcus aureus and Bacillus subtilis with MIC value 12.5mg/L and 25mg/L. Compounds 1－4 were isolated from the fungus for the first time.
short communications
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  Three newly recorded species of Veronaea and Wardomyces from China

  Mycosystema. 2012, 31(1): 133-135.

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  Three new records of Veronaea coprophila, Wardomyces anomalus and Wardomyces humicola to China from soil of Yellow River source area, were described and illustrated. Specimens studied were deposited in the Plant Pathology Herbarium of Shandong Agricultural University (HSAUP).
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  Three tropical polypores (Basidiomycota) new to China

  Mycosystema. 2012, 31(1): 136-141.

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  Lignicolous fungi are very important functional organisms in forest ecosystem and play a key role in materials circulation. Diversity and taxonomic investigations on lignicolous fungi, especially from tropical zones in China, were carried out in recent years. During the investigations in Hainan and Yunnan provinces, three polypore species were identified as Junghuhnia crustacean, Phellinus tropicalis and Polyporus blanchettianus. They belong to the Meruliaceae, Hymenochataceae and Polyporaceae respectively, and are newly recorded from China. Descriptions and illustrations of these species are given based on the materials collected from China, and the relationships between their similar species are discussed.
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  Cloning and expression analysis of a polyketide synthase gene vv-alb from the edible straw mushroom, Volvariella volvacea

  Mycosystema. 2012, 31(1): 142-149.

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  Polyketides are secondary metabolites that exhibit a broad range of biological activities. The key enzyme involved in biosynthesis is polyketide synthase (PKS). We report the cloning of a putative PKS gene (vv-alb) by nested PCR using degenerate primers and chromosome walking from the edible straw mushroom, Volvariella volvacea. The gene vv-alb is 6,704bp in size and contains a 6,279bp coding region (encoding 2,093 amino acid residues) and eight introns. The encoded product of the vv-alb gene contains four functional domains: ketosynthase (KS), acyl transferase (AT), thioesterase (TE) and acyl carrier protein (ACP), the typical characteristics of PKS. Real-time quantitative PCR analysis revealed that vv-alb expression was 5-10-fold higher in the stipe region as compared with the vegetative mycelium, primordia and pileus.