A new <Emphasis Type="Italic">Coniosporium</Emphasis> species from historical marble monuments

The aim of this investigation was to use morphological and molecular genetic techniques to characterize strains of newly discovered biodeteriorative fungal species originating from antique marble monuments in the Mediterranean basin (Side, Antalya, Turkey). The strains studied were isolated from the lid of a sarcophagus in the Side Museum. Black colonies were picked up and purified by one to several transfers onto culture media. Their morphology was characterized, and DNA was extracted, and amplified. Novelty of the selected species is supported by sequencing of the small ribosomal subunit (18 S) and internal transcribed spacer (ITS) regions. The black meristematic species is placed in the genus Coniosporium. Judging from SSU phylogeny data, this species is supposed to belong to the Chaetothyriales. Analysis of nucleotide sequences did not show a degree of similarity to any sequence in Genbank that would allow to assign it to one of the already existing species. The new species was denominated as C. sümbülii.     

Molecular taxonomy and biodiversity of rock fungal communities in an urban environment (Vienna, Austria)

The diversity of fungal communities on three different historical monuments in the city of Vienna (Austria) was analyzed and compared to the fungal diversity of microfungi on rock in the original quarry located in a rural area (Zogelsdorf, Austria). The fungal strains isolated were characterized by morphology and the complete rock fungal community was identified based on molecular data, that is, by sequencing parts of the small ribosomal subunit (18S) and internal transcribed spacer region 1 (ITS1). The genera Coniothyrium, Epicoccum and Phoma were found to be dominant {on} monument and rock surfaces. Additionally, black yeasts such as Exophiala species and microcolonial fungi like Sarcinomyces and Coniosporium which hitherto were regarded as typical rock inhabitants in semi-arid environments are frequently found on all rock surfaces in Vienna. The biodiversity of the fungi in the urban environment was much higher than on the same rock type in a rural environment, this difference can be attributed to the elevated organic pollution in the city.     

Phylogenetic analysis and screening of antimicrobial and cytotoxic activities of moderately halophilic bacteria isolated from the Weihai Solar Saltern (China)

A total of 45 moderately halophilic bacteria was isolated from sediment and saline water collected from the Weihai Solar Saltern (China). The phylogenetic position of all the isolated strains was determined by 16S rRNA sequencing. The halophilic strains were tested for their antimicrobial activity. Cytotoxicity assay was performed to determine which of the halophilic strains could inhibit proliferation of human hepatocellular carcinoma Bel 7402 cells. Our results showed that all of the isolated 45 strains displayed moderately halophilic characteristics. Phylogenetic analysis indicated that 17 of the isolated strains were related to the phylum Firmicutes and belonged to four genera, Bacillus, Halobacillus, Planococcus and Salinicoccus. The other strains identified as genus of Halomonas belonged to phylum γ-Proteobacteria. Most of the halophilic bacterial strains showed potent activities against Gram-positive bacteria, human pathogenic fungi and plant pathogenic fungi. In addition, the crude extracts from 14 halophilic bacterial strains showed cytotoxic activity against tumor cells Bel 7402, and five of them showed remarkable activities with IC₅₀ less than 40 μg ml⁻¹. Our results suggest that the moderately halophilic bacteria may be developed as promising sources for the discovery of novel bioactive substances.     

剑叶龙血树内生真菌的分离及体外抑菌活性筛选

为研究剑叶龙血树内生真菌资源多样性,初步探讨和筛选具有抑菌活性的特异性菌株以及进一步开发剑叶龙血树内生真菌的抗菌活性化合物.该文采用植物组织分离法从剑叶龙血树茎和叶中分离内生真菌,对内生真菌进行液体发酵7 d,经乙酸乙酯萃取后制得粗提物,并采用牛津杯扩散法,以10种常见病原菌和5种临床耐药菌为靶标检测其发酵粗提物的抑菌...     

Robusta coffee beans post-harvest microflora: Lactobacillus plantarum sp. as potential antagonist of Aspergillus carbonarius

Coffee contamination by ochratoxigenic fungi affects both coffee quality as well as coffee price with harmful consequences on the economy of the coffee exporting countries for whom which is their main source of income. Fungal strains were isolated from coffee beans and identified as black Aspergilli. Ochratoxigenic moulds like Aspergillus carbonarius were screened and selected for detailed studies. Also lactic acid bacteria (LAB) were isolated from silage coffee pulp and their antifungal activity was tested on dual-culture agar plate. Ten of the isolated LAB demonstrated antifungal effect against A. carbonarius. API 50 CH and APIZYM were used to perform phenotypic identification. 16S rDNA sequencing was made to confirm the results.     

川蔓藻内生及根际细菌多样性与抑菌活性研究

该研究采用稀释涂布法结合形态观察、16S rRNA基因序列分析,对广西北海川蔓藻(Ruppia maritima)内生及根际细菌的物种多样性进行了研究,并采用琼脂扩散法和光度计法分析了其粗提物抑制马尔尼菲青霉菌活性。结果表明:从川蔓藻中分离到可培养内生细菌26株,根际可培养细菌31株。分别将内生细菌归属为10科12属13种,根际分离出细菌归属为9科14属19种,其中5株根际细菌可能为潜在新种。获得8株细菌对马尔尼菲青霉菌有抑制活性,总阳性率为25.0%。其中,菌株BGMRC 2015、BGMRC 2059、BGMRC 2043的粗提物表现出较强的抑制马尔尼菲青霉菌效果,其MIC分别为(1.800±0.045)、(1.881±0.061)、(1.604±0.021)mg·m L~(-1)。川蔓藻中可培养细菌具有较高的物种多样性,蕴藏着丰富的新物种资源,且富含抑菌活性良好的菌株。     

Fusarium fractiflexum sp. nov. and two other species within theGibberella fujikuroi species complex recently discovered in Japan that form aerial conidia in false heads

Morphological and molecular phylogenetic analyses were conducted on 12 strains ofFusarium, deposited in MAFF asF. subglutinans (≡F. moniliforme var.subglutinans≡F. sacchari var.subglutinans) orFusarium sp. because they formed aerial conidia in false heads in the dark. These strains were resolved as three distinct species within theGibberella fujikuroi species complex. A new species,F. fractiflexum, and two species new to Japan,F. circinatum andF. concentricum, are described and illustrated and their morphological features are discussed.Fusarium fractiflexum, isolated from diseased yellow leaf spots ofCymbidium spp., is differentiated from other fusaria based on its yellowish colonies and aerial conidia formed in false heads in the dark and in zigzag-like conidial chains under black light. Japanese strains ofF. circinatum also formed elongate, coiled sterile hyphae. Phialidic aerial conidia with a pointed apex and a wedgeshaped base were found inF. concentricum cultured under black light and represent a new diagnostic character of the species, in addition to colonies with alternating concentric rings when cultured on PDA. Based on DNA sequences of the β-tubulin gene and two other loci, strains ofF. fractiflexum were resolved phylogenetically as members of the Asian clade of theG. fujikuroi species complex. In addition, Japanese strains ofF. circinatum andF. concentricum were phylogenetically identical to the ex-type strains.     

两种栽培型广藿香内生真菌群落组成变化

为探索内生真菌与广藿香互作间对宿主活性成分形成机制的影响,该研究以成分差异较大的牌香和湛香为对象,采用传统形态学方法对所获菌株归类,通过真菌通用引物ITS1/ITS4扩增菌株rDNA-ITS序列,鉴定其分类地位并研究其多样性。结果表明：(1)用PDA和LBA培养基对苗期、分枝期和成株期广藿香茎叶组织块进行内生真菌分离,共获得3 070株菌株,其中牌香(PX)分离出1 624株,鉴定出1 319株,分属于36属;湛香(ZX)分离出1 446株,鉴定出994株,分属于33属。牌香分离出7种特有内生真菌,分别为香柱菌(Epichloe typhina)、盘长孢状刺盘孢菌(Colletotrichum gloeosporioides)、座腔孢菌(Botryosphaeria sp.)、丝核菌(Rhizoctonia sp.)及截盘多毛孢菌(Truncatella sp.),并首次分离到疫霉菌(Phytophthora sp.)和指疫霉菌(Sclerophthora sp.),这2种菌属于卵菌门内生菌。湛香分离出拟青霉菌(Paecilomyces sp.)和尾孢菌(Cercospora sp.)...     

The Lichen Connections of Black Fungi

Many black meristematic fungi persist on rock surfaces—hostile and exposed habitats where high doses of radiation and periods of desiccation alternate with rain and temperature extremes. To cope with these extremes, rock-inhabiting black fungi show phenotypic plasticity and produce melanin as cell wall pigments. The rather slow growth rate seems to be an additional prerequisite to oligotrophic conditions. At least some of these fungi can undergo facultative, lichen-like associations with photoautotrophs. Certain genera presenting different lifestyles are phylogenetic related among the superclass Dothideomyceta. In this paper, we focus on the genus Lichenothelia, which includes border-line lichens, that is, associations of melanised fungi with algae without forming proper lichen thalli. We provide a first phylogenetic hypothesis to show that Lichenothelia belongs to the superclass Dothideomyceta. Further, culture experiments revealed the presence of co-occurring fungi in Lichenothelia thalli. These fungi are related to plant pathogenic fungi (Mycosphaerellaceae) and to other rock-inhabiting lineages (Teratosphaeriaceae). The Lichenothelia thallus-forming fungi represent therefore consortia of different black fungal strains. Our results suggest a common link between rock-inhabiting meristematic and lichen-forming lifestyles of ascomycetous fungi.     

块菌(松露)宿主可培养内生菌群落结构与多样性研究

探究四川凉山彝族自治州块菌主产区华山松内生菌群的结构及多样性。在会东县选取4个点(新田乡、新云乡、淌塘镇、雪山乡)的块菌宿主华山松的根、茎、叶为实验材料,通过不同培养基分离样品根、茎、叶的内生细菌、真菌、放线菌,DNA分子鉴定分离菌株的种属,最终分离得到细菌46株,其中欧文氏菌属(Erwinia)5株,沙门氏菌属(Salmonell)1株,杆菌属(Bacillus)22株,葡萄球菌属(Staphylococcus)2株,假单胞菌属(Pseudomonas)2株,类芽胞杆菌属(Paenibacillus)2株,布丘氏菌属(Buttiauxella)2株,肠杆菌属(Enterobacte)3株,爱文氏菌属(Ewingella)1株,泛菌属(Rahnella)1株,拉恩氏菌属(Rahnella Izard)2株,其他属3株;真菌19株,均为子囊菌门(Ascomycota),其中青霉属(Penicillium)4株,疱霉属(Phoma)1株,子囊菌门未知菌1株,篮状菌属(Cladosporium)1株,分枝孢子菌属(Sydowia)1株,曲霉属(Aspergillus)1株,其他属10株;放线菌33株,为链霉菌属(Streptomyces)22株,短小杆菌属(Curtobacterium)2株,短杆菌属(Brevibacterium)1株,其他属8株。研究结果表明,来自不同地点、不同植株部位、不同培养基分离得到的块菌宿主华山松内生菌有差异,证实微生物在土壤、块菌、宿主植物之间有着复杂的相互作用,为块菌的人工栽培提供参考。     

带叶兜兰种子原地共生萌发及有效菌根真菌的分离与鉴定

为获得带叶兜兰(Paphiopedilum hirsutissimum)种子萌发的共生真菌,采用原地共生萌发技术获得了2株自然萌发的小幼苗,并分离和筛选出了有效的种子萌发共生菌——瘤菌根菌(Epulorhiza sp.)。为验证分离菌株对带叶兜兰种子萌发的有效性,将Phs34号菌株与带叶兜兰种子在灭菌后的原生境基质上进行室内共生萌发试验,结果表明,经过6周的培养,对照组没有观察到种子的萌发;接菌的种子胚明显膨大,突破种皮,形成原球茎,平均萌发率为(58.35±3.41)%。这表明分离得到的瘤菌根菌能促进带叶兜兰的种子萌发。     

Diversity and annual fluctuations of culturable airborne fungi in Athens, Greece: a 4-year study

The diversity and the abundance of the culturable airborne fungi have been studied by a volumetric method in the city of Athens, for a period of 4 years. A total of 6,600 plates were exposed during 562 calendar days, and 70,583 colonies of fungi have been recovered and studied in detail. One hundred and forty-eight species in fifty-four genera of filamentous fungi were identified. A total of three hundred and twenty strains were isolated and maintained as reference material. The annual mean concentration of the total fungi was 538, 640, 694 and 638 CFU/m³, and the concentration range, 25–2,435, 117–2,822, 122–2,201 and 116–2,590 CFU/m³ for each year, respectively. There is no statistically significant year-to-year variation in the distribution patterns and in the annual mean concentrations of the total fungi. The diversity and the abundance of the total fungi and of the dominant genera Cladosporium, Aspergillus and Alternaria were increased, whereas those of Penicillium decreased during the warm months of each year. The majority of the species are newly reported as airborne from Greece. Also, 19 genera and 93 species are totally new records for this country. The species Acrodontium virellum, Aspergillus aculeatus, A. tubingensis, Circinella minor, C. umbellata, Cladosporium breviramosum, C. malorum, Drechslera tetramera, Paecilomyces crustaceus, Petriella guttulata, Rutola graminis and Sporotrichum pruinosum are reported as airborne for the first time worldwide.     

金荞麦和苦荞麦抗菌活性内生真菌的筛选及鉴定

从药用植物金荞麦和苦荞麦的根、茎、叶、花中分离到62株内生真菌,并以金黄色葡萄球菌(Staphylococcus aureus)、大肠杆菌(Escherichia coli)、枯草芽孢杆菌(Bacillus subtilis)[CMCC(B)63501]、小麦赤霉病菌(Fusarium graminearum)、黄瓜枯萎病菌(Fusarium oxysporum f.sp.cu-cumerinum)和绵腐病菌(Pythium aphanidermatum)6种微生物为指示菌对分离到的内生真菌进行抗菌活性检测。结果发现,分离的内生真菌菌株KQH-01、KQH-02和JQY-1的发酵醇提取物具有较好的抑菌活性。形态学特征和分子鉴定确定菌株KQH-01为炭角菌属(Xylaria sp.)真菌,菌株KQH-02为球毛壳菌(Chaetomium globosum),菌株JQY-1为葡萄座腔菌(Botryosphaeria dothidea)。     

Screening for Ligninolytic Enzyme Production by Diverse Fungi from Tunisia

Summary This work represents the first report on the ability of autochthonous fungi from Tunisia to produce ligninolytic enzymes. Three hundred and fifteen fungal strains were isolated from different Tunisian biotopes. These fungal strains were firstly screened on solid media containing Poly R-478 or ABTS as indicator compounds that enabled the detection of lignin-modifying enzymes as specific color reactions. Of the 315 tested strains, 49 exhibited significant ABTS-oxidation activity expressed within the first week of incubation and only 18 strains decolorized the Poly R-478. Liquid cultivations and laccase, manganese peroxidase and lignin peroxidase activity assays of positive strains confirmed that eight efficient enzyme producers were found in the screening. These strains were attributed to the most closely related species using PCR amplification and sequencing of the internal transcribed spacer ‘ITS’ regions of the ribosomal DNA. The identification results showed fungal genera such as Oxyporus, Stereum and Trichoderma which have been only rarely reported as ligninolytic enzyme producers in the literature. Culture conditions and medium composition were optimized for the laccase producer Trametes trogii CTM 10156. This optimization resulted in high laccase production, 367 times more than in non-optimized conditions and which reached 110 U ml^-1 within 15 days of incubation.     

高磷土壤中丛枝菌根真菌的分离鉴定

【背景】丛枝菌根(arbuscular mycorrhiza,AM)真菌具有广泛的寄主范围、环境适应性和优良的植物促生能力。然而,土壤的高磷水平严重抑制了AM真菌生长及AM形成。【目的】分离鉴定出耐较高有效磷含量的华南土著AM真菌菌株,为菌根学研究工作提供新颖材料。【方法】采用经典形态学和分子系统学方法鉴定高磷土壤中AM真菌。【结果】从有效磷含量为53-131 (平均值±标准差为88.2±17.6) mg/kg的根区土壤中鉴定出7属25种AM真菌,包括无梗囊霉属(Acaulospora) 12种、球囊霉属(Glomus) 7种、隔球囊霉属(Septoglomus) 2种、近明球囊霉属(Claroideoglomus) 1种、根孢囊霉属(Rhizophagus) 1种、硬囊霉属(Sclerocystis) 1种和类球囊霉属(Paraglomus) 1种,其中幼套近明球囊霉(Claroideoglomus etunicatum)和蜜色无梗囊霉(Acaulospora mellea)是优势种。在(87.7±8.0) mg/kg的高磷水平下,AM真菌仍能形成丛枝和泡囊。但当有效磷含量达到(99.7±1.2) mg/kg时,菌根侵染率和丛枝丰度显著下降,但仍能够形成泡囊。【结论】从广州市南沙区有效磷含量为(88.2±17.6) mg/kg的耕地植物根区土壤中,鉴定出具有耐高磷潜力的7属25种AM真菌,幼套近明球囊霉和蜜色无梗囊霉等分离株可作为后续高磷抑制机制解析及耐高磷AM真菌菌剂研发工作的试验菌株。     

Fungal Epiphytes and Endophytes of Coffee Leaves (Coffea arabica)

Plants harbor diverse communities of fungi and other microorganisms. Fungi are known to occur both on plant surfaces (epiphytes) and inside plant tissues (endophytes), but the two communities have rarely been compared. We compared epiphytic and endophytic fungal communities associated with leaves of coffee (Coffea arabica) in Puerto Rico. We asked whether the dominant fungi are the same in both communities, whether endophyte and epiphyte communities are equally diverse, and whether epiphytes and endophytes exhibit similar patterns of spatial heterogeneity among sites. Leaves of naturalized coffee plants were collected from six sites in Puerto Rico. Epiphytic and endophytic fungi were isolated by placing leaf pieces on potato dextrose agar without and with surface sterilization, respectively. A total of 821 colonies were isolated and grouped into 131 morphospecies. The taxonomic affinities of the four most common nonsporulating fungi were determined by sequencing the nuclear ribosomal internal transcribed spacer (ITS) region: two grouped with Xylaria and one each with Botryosphaeria and Guignardia. Of the most common genera, Pestalotia and Botryosphaeria were significantly more common as epiphytes; Colletotrichum, Xylaria, and Guignardia were significantly more common as endophytes. Suprisingly, more morphospecies occurred as endophytes than as epiphytes. Differences among sites in number of fungi per plant were significant. Thus epiphytic and endophytic communities differed greatly on a single leaf, despite living only millimeters apart, and both communities differed from site to site. Significant correlations between occurrence of fungal morphospecies suggested that fungi may have positive or negative effects on their neighbors. This is the first quantitative comparison of epiphytic and endophytic fungal floras in any plant, and the first to examine endophytic fungi or epiphytic fungi in leaves of coffee, one of the world’s most valuable crops.     

Toxicity of a <Emphasis Type="Italic">Bacillus thuringiensis israelensis</Emphasis>-like strain against <Emphasis Type="Italic">Spodoptera frugiperda</Emphasis>

Bacillus thuringiensis (Bt) Berliner is a promising agent for microbial control of agriculturally and medically important insects. This study aimed at searching for Bt strains encoding Cry proteins that act more efficiently against fall armyworm. Thirty Bt strains were isolated from soil samples in Pernambuco State and evaluated through bioassays. Among these, strain I4A7 was the most efficient against the fall armyworm, Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae), and thus it was characterized by biochemical sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and molecular (polymerase chain reaction (PCR) and sequencing reaction) methods. The protein pattern of this strain on a SDS–PAGE was similar to that of B. thuringiensis israelensis (Bti). Moreover, I4A7 cry DNA sequence showed high identity (99–100%) to genes cry4Aa, 4Ba, 10Aa, 11Aa, cyt1Aa and cyt2B from Bti. The toxicity of the newly isolated Bti-like strain upon S. frugiperda should be considered as this strain might be used in combination with other Bt strains, such as B. thuringiensis var. kurstaki (Btk). Handling Editor: Helen Roy.     

Isolation and characterization of entomopathogenic fungi from hazelnut-growing region of Turkey

Although Turkey is the first among all hazelnut-producing countries, yield per unit area of this crop is low in comparison to other countries, mainly because many insect species seriously damage hazelnut trees and their fruit. To find effective and safe biocontrol agents, we conducted a survey study to isolate entomopathogenic fungi from the hazelnut-growing region of Turkey and characterized the isolated strains in detail. In addition, we determined the effectiveness of seven selected strains from this region against Melolontha melolontha (Coleoptera: Scarabaeidae) which is one of the most serious pests of hazelnut. In 2006 and 2007, 301 soil samples were collected randomly and analyzed for presence of entomopathogenic fungi using the Galleria bait method. Entomopathogenic fungi were found to occur in 20.59% of the soil samples studied. Based on morphology, ITS sequence and partial sequencing of the 18S (SSU rDNA) and EF1-α genes, the isolates were identified as Metarhizium anisopliae var. anisopliae, Metarhizium sp., Beauveria bassiana, Beauveria cf. bassiana, Isaria fumosorosea and Evlachovaea sp. Metarhizium anisopliae var. anisopliae was isolated from 34 sites and was the most frequent and abundant entomopathogenic species recovered. All the isolates tested were pathogenic to M. melolontha. M. anisopliae var. anisopliae KTU-27 and Evlachovaea sp. KTU-36 produced the highest insecticidal activity (86.6%) within 15 days after inoculation. Our results suggest that entomopathogenic fungi could be good biocontrol agents against M. melolontha, and are discussed with respect to ecology of fungi in relation to habitat in order to evaluate biocontrol potential of these isolates. This is the first study of the distribution of entomopathogenic fungi in the hazelnut-growing region of Turkey and of their pathogenicities against M. melolontha.     

粮食上木霉菌的分离鉴定及其生防效果

【背景】粮食在生长和收储期极易受到病原真菌或产毒真菌的污染,造成严重的损失。众多实践证明木霉属(Trichoderma)可以有效防治植物病原真菌。【目的】鉴定和筛选能有效抑制粮食常见危害真菌的木霉生防菌株,开发生防菌剂,保障粮食生产安全。【方法】从粮食上分离筛选出35株木霉,通过多基因系统发育分析和形态学观察方法进行菌种鉴定,利用平板对峙试验筛选出对粮食常见危害真菌有抑制作用的菌株。【结果】35株木霉分属于8个种,分别为非洲哈茨木霉(Trichodermaafroharzianum)、类棘孢木霉(Trichodermaasperelloides)、 Trichoderma amoenum、近深绿木霉(Trichoderma paratroviride)、Trichoderma obovatum、长枝木霉(Trichoderma longibrachiatum)、东方木霉(Trichodermaorientale)和深绿木霉(Trichodermaatroviride)。对峙试验结果表明,这8种木霉对于粮食上分离到的10种危害真菌均具有较好的抑制效果。非洲哈茨木霉(T.afroharzi...     

黄花倒水莲内生真菌生物活性评价及HNLF-44菌株鉴定

为充分开发黄花倒水莲(Polygala fallax)的内生真菌资源，获得具有抗植物病原真菌、抗氧化活性的内生真菌，该文以黄花倒水莲内生真菌为研究对象，使用平板对峙法检测内生真菌对6种植物病原真菌的抑菌活性，测定内生真菌发酵液的DPPH清除自由基能力和总还原能力，评价内生真菌的抗氧化活性，并对具有强抑菌活性和抗氧化活性的菌株进行形态和ITS鉴定。结果表明：(1)黄花倒水莲内生真菌中有2株内生真菌对香蕉专化尖孢镰刀菌、柑橘树脂病菌、叶点霉菌、香蕉具条叶斑病菌、茄病镰刀菌、三七根腐病菌具有明显的抑菌活性，抑菌率在50.3%～91.4%之间，其中HNLF-5对柑橘树脂病菌的抑菌率为73.2%,HNLF-44对香蕉专化尖孢镰刀菌抑菌率为91.4%。(2)内生真菌发酵液具有良好的抗氧化活性，DPPH清除率均在80%以上，总还原能力吸光值范围为0.279 2～0.748 8。(3)HNLF-44菌株为链格孢属真菌。该研究表明，药用植物黄花倒水莲内生真菌具有较好的生物活性，为后续从黄花倒水莲内生真菌中挖掘潜在新型抑菌活性和抗氧化活性物质奠定了基础。