A new species from tropical soils,Eupenicillium tropicum

Forty-three strains of Eupenicillium tropicum sp. nov. were isolated from soils collected in India, Costa Rica and Galapagos, Ecuador. The species is characterized by biverticillate penicilli, slightly rough, subglobose to ovate conidia, brownish cleistothecia that become brown-gray with age, and ascospores with two equatorial flanges and slightly roughened valves. All strains produced a large number of indole alkaloids, and many types of unknown secondary metabolites with characteristic chromophores were produced by a majority of strains. Eupenicillium tropicum is morphologically most similar to E. shearii, but based on ITS-LSU sequences, is most closely related to Penicillium citrinum, P. sartoryi and P. westlingii. Eupenicillium shearii strains consistently produce paxillin, paspalinine and shearinins, while the latter three penicillia all produce citrinin consistently.     

<Emphasis Type="Italic">Hirsutella proturicola</Emphasis> sp. nov. isolated from a proturan, <Emphasis Type="Italic">Baculentulus</Emphasis> densus (Protura,Hexapoda)

A new species of Hirsutella, H. proturicola, isolated from a subterranean proturan (Baculentulus densus; Protura, Hexapoda), is described and illustrated. Hirsutella proturicola is characterized by producing monoblastic phialides of 24–51.5 × 2.5–5 μm with a slightly roughened neck, fusiform and curved conidia of 9–18 × 2.5–4 μm that have a truncate base and a papillate projection often capped with sheath-like mucilage, and pluricellular, globose to subglobose chlamydospores of 21–48 × 21–41.5 μm. This species is morphologically and phylogenetically close to H. rostrata, an acaropathogenic species, but can be distinguished from the size of the phialides and the size and shape of the conidia.     

<Emphasis Type="Italic">Penicillium svalbardense</Emphasis>, a new species from Arctic glacial ice

During investigations of mycobiota in the coastal Arctic polythermal glaciers, different species of the ubiquitous genus Penicillium were isolated from the extreme subglacial environment. A group of Penicillium strains was obtained that did not belong to any known Penicillium species. This species was isolated in high numbers from the Kongsvegen subglacial ice and was not detected in the surrounding environment. A detailed analysis of secondary metabolite profiles, physiological and morphological characteristics, and partial β-tubulin gene sequences showed that the proposed new species Penicillium svalbardense is closely related but not identical to Penicillium piscarium and Penicillium simplicissimum. It differs in the production of secondary metabolites and in the morphological features of conidia and penicilli, and it is therefore described as a new species.     

Three newAspergillus species isolated from clinical sources as a causal agent of human aspergillosis

Three new species ofAspergillus isolated from clinical sources in China are described and illustrated:A. beijingensis, A. qizutongii andA. wangduanlii. The first species is characterized by spreading colonies, yellow to grayish green conidial heads, smooth-walled conidiophores with a clavate vesicle, uniseriate aspergilla and nearly globose, micro-verrucose conidia. The second is characterized by spreading colonies, olive-yellow conidial heads, conspicuously roughened conidiophores with a flask-shaped vesicle, uniseriate but often secondarily proliferating aspergilla and globose, smooth conidia. The third is characterized by rapidly growing colonies, dull green conidial heads, smooth to irregularly roughened conidiophores which are often surrounded by coiled hyphae in the basal part and are terminally swollen into a globose or irregular shaped vesicle, uniseriate aspergilla and globose, micro-verrucose conidia.     

Morphological and molecular characterisation of Penicillium roqueforti and P. paneum isolated from baled grass silage

The morphological and molecular features of Penicillium roqueforti and P. paneum isolated from baled grass silage were characterised. A total of 315 isolates were investigated, comprising 237 P. roqueforti and 78 P. paneum isolates randomly selected from more than 900 Penicillium colonies cultured from bales. The macromorphological features of both species broadly agreed with the literature, but the micromorphological features differed in some respects. When observed using SEM, P. roqueforti and P. paneum had finely roughened conidia, and conidiophores, phialides and conidia of P. paneum were each larger than those of P. roqueforti. Based on the phylogenetic analysis of partial sequences of β-tubulin and acetyl co-enzyme A (CoA) synthetase genes, P. roqueforti and P. paneum isolates were found to be monophyletic species.     

Penicillium nodositatum Valla,a new species inducing myconodules on Alnus roots

Penicillium nodositatum Valla, a new species of Penicillium, able to induce nodule formation on the root system of two alder species, is described in this paper. It belongs to the subgenusBiverticillium ser.Islandica. Typical distinctive features of this species are canary to chrome yellow mycelium, ampulliform to acerose phialids and ellipsoid, echinulate conidia. The taxonomic position ofP. nodositatum and a comparison with another myconodule inducing species (P. janczewskii Zaleski) are discussed.     

Use of the Weibull model to describe inactivation of dry‐harvested conidia of different Penicillium species by ethanol vapours

Aims: This study aimed at modelling the effect of ethanol vapours, in the range 0·7–7·5 kPa, on the inactivation of dry‐harvested conidia of Penicillium chrysogenum, Penicillium digitatum and Penicillium italicum. Methods and Results: Survival curves were modelled by a Weibull model: log (N/N₀) = ?1/2·303 (t/α)^β. The shape parameter β was different from one in all cases, indicating that the classical first‐order kinetics approach is the exception rather than the rule. Survival curves exhibited upward concavity (β < 1) with the notable exception of P. chrysogenum at ethanol vapour pressures 0·7 and 1·5 kPa. The scale parameter α (h) varied greatly depending on the ethanol vapour pressure and on the species. Conclusions: For safety reasons, it is recommended not to exceed an ethanol vapour pressure of 3·3 kPa. At 2·8 kPa, more than 4 log₁₀ reductions in viable conidia were achieved for all the species after 24‐h exposure. Significance and Impact of the Study: Ethanol has GRAS status in the USA and represents an interesting alternative to fungicides. The effectiveness of ethanol vapours to inactivate dry‐harvested conidia of some Penicillium was demonstrated in this study.     

Biotic effect of fungal communities inhabiting grapevine phyllosphere on <Emphasis Type="Italic">Phoma negriana</Emphasis>

The effect of 15 fungi species most frequently inhabiting grapevine shoots on the growth of Phoma negriana was examined. Particular species-components of the fungal community were tested in vitro against the pathogen, using the biotic series method. The majority of fungi species inhibited P. negriana growth and the growth of only 3 species from the community, i.e. Alternaria alternata Keiss., Botrytis cinerea Pers. and Phomopsis viticola, was limited by the pathogen. Fungi Trichoderma were found the most effective against P. negriana. They completely degraded the hyphae and conidia of P. negriana after 20–22 days of growth in two-organism cultures. The summary biotic effects of fungal communities from grapevine were generally favourable to the P. negriana growth and only in one year of study they suppressed the pathogen development.     

Prevalence,geographic distribution and phylogenetic relationships among cryptic species of Plasmopara viticola in grape‐producing regions of Georgia and Florida,USA

Previous phylogenetic studies of the grape downy mildew pathogen, Plasmopara viticola, revealed five cryptic species in eastern North America that differed in their host range and geographic distribution. Preliminary comparative studies also documented differences in temperature responses during infection between certain cryptic species, indicating the biological relevance of knowing which cryptic species of the pathogen are present in a given region. However, limited information is available regarding the presence, prevalence and dynamics of cryptic species of P. viticola in the southeastern United States. Here, 301 P. viticola isolates obtained from cultivated grape species in five distinct grape‐growing regions of Georgia and Florida were subjected to cleaved amplified polymorphic sequence analysis and multilocus sequencing (internal transcribed spacer region of the rDNA, actin and β‐tubulin) to identify cryptic species and infer phylogenetic relationships. Three cryptic species, P. viticola clade aestivalis (Pva), clade vinifera (Pvv) and clade vulpina (Pvu), were identified in Georgia, whereas two, Pva and Pvv, were found in Florida; all three cryptic species are reported here for the first time in Georgia, whereas Pva is reported for the first time in Florida. Pva was the most prevalent cryptic species (72.1% of isolates) and was distributed widely from the North Georgia Mountains to Mid‐Florida, whereas Pvv (27.2%) and Pvu (0.7%) were found only in the Coastal Plain region of the two states. Interestingly, Pvu was obtained from French American hybrid Blanc du Bois and could be subcultured on Vitis vinifera Chardonnay, suggesting a broader host range than only the wild species Vitis vulpina reported previously.     

<Emphasis Type="Italic">Metschnikowia viticola</Emphasis> sp. nov., a new yeast species from grape

Two yeast strains, producing needle-shaped ascospores under suitable conditions, were isolated from grapes grown in Hungary. Based on these two strains, Metschnikowia viticola (type strain NCAIM Y.01705, CBS 9950, JCM 12561) is proposed as a new yeast species. Considering its phenotypic features, the restriction fragment patterns of 18S rDNA and the sequence of the D1/D2 domain of 26S rDNA, the proposed new species is closely related to Candida kofuensis.     

Grapevine VpPR10.1 functions in resistance to Plasmopara viticola through triggering a cell death‐like defence response by interacting with VpVDAC3

As one of the most serious diseases in grape, downy mildew caused by Plasmopara viticola is a worldwide grape disease. Much effort has been focused on improving susceptible grapevine resistance, and wild resistant grapevine species are important for germplasm improvement of commercial cultivars. Using yeast two‐hybrid screen followed by a series of immunoprecipitation experiments, we identified voltage‐dependent anion channel 3 (VDAC3) protein from Vitis piasezkii ‘Liuba‐8’ as an interacting partner of VpPR10.1 cloned from Vitis pseudoreticulata ‘Baihe‐35‐1’, which is an important germplasm for its resistance to a range of pathogens. Co‐expression of VpPR10.1/VpVDAC3 induced cell death in Nicotiana benthamiana, which accompanied by ROS accumulation. VpPR10.1 transgenic grapevine line showed resistance to P. viticola. We conclude that the VpPR10.1/VpVDAC3 complex is responsible for cell death‐mediated defence response to P. viticola in grapevine.     

The sequence of the isoepoxydon dehydrogenase gene of the patulin biosynthetic pathway in <Emphasis Type="Italic">Penicillium</Emphasis> species

Interest in species of the genus Penicillium is related to their ability to produce the mycotoxin patulin and to cause spoilage of fruit products worldwide. The sequence of the isoepoxydon dehydrogenase (idh) gene, a gene in the patulin biosynthetic pathway, was determined for 28 strains representing 12 different Penicillium species known to produce the mycotoxin patulin. Isolates of Penicillium carneum, Penicillium clavigerum, Penicillium concentricum, Penicillium coprobium, Penicillium dipodomyicola, Penicillium expansum, Penicillium gladioli, Penicillium glandicola, Penicillium griseofulvum, Penicillium paneum, Penicillium sclerotigenum and Penicillium vulpinum were compared. Primer pairs for DNA amplification and sequencing were designed from the P. griseofulvum idh gene (GenBank AF006680). The two introns present were removed from the nucleotide sequences, which were translated to produce the IDH sequences of the 12 species for comparison. Phylogenetic relationships among the species were determined from rDNA (ITS1, 5.8 S, ITS2 and partial sequence of 28S rDNA) and from the idh nucleotide sequences minus the two introns. Maximum parsimony analysis showed trees based on rDNA and idh sequences to be congruent. It is anticipated that the genetic information obtained in the present study will aid in the design of probes, specific for patulin biosynthetic pathway genes, to identify the presence of these mycotoxigenic fungi. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

<Emphasis Type="Italic">Dictyochaeta multifimbriata</Emphasis>, a new species from Taiwan

A new species of the dematiaceous hyphomycete genus Dictyochaeta was collected on a dead twig in Taiwan. It is characterised by solitary conidiophores and reniform, non-septate conidia with several setulae at each end. The species is compared with other species having non-septate, setulate conidia and is described as Dictyochaeta multifimbriata.     

Aspergillus osmophilus sp. nov., and a new teleomorph for A. proliferans

A new species of Aspergillus and a new teleomorph for A. proliferans, both isolated from cereals in Iran, are described using morphological and molecular data. A combined sequence dataset of the ITS region, partial β-tubulin and partial calmodulin genes resolved the relationships of members of section Aspergillus largely in concordance with morphological traits of ascospores. Aspergillus osmophilus sp. nov. is differentiated from the closest species, A. xerophilus by possessing larger ascospores, conidia and associated fruiting bodies. Both species are strongly xerophilic and possess ascospores with lobate-reticulate convex surfaces. The newly discovered teleomorph for A. proliferans is characterized by delicately roughened ascospores with a shallow or distinct furrow and finely roughened to irregular equatorial crests.     

Hydrophobicity and surface electrostatic charge of conidia of the mycoparasitic <Emphasis Type="Italic">Trichoderma</Emphasis> species

The present investigation was done to understand the fungal-fungal interactions mechanisms based on level of nonspecific adhesion of a potential fungal mycoparasite (Trichoderma) to their fungal host (Macrophomina phaseolina). The relative cell surface hydrophobicity (CSH) and cell surface electrostatic charge (CSEC) of 29 isolates of Trichoderma species, analyzed by bacterial adhesion to hydrocarbon (BATH), hydrophobic interaction chromatography (HIC), microelectrophoresis and contact angle, revealed a large degree of variability. CSH and CSEC of conidia depended on culture age, pH and temperature. Maximum CSH and CSEC were recorded in 25–28 °C range, and both declined significantly with increasing temperature. Isolate Trichoderma hazianum (Th)-23/98 expressed surface hydrophobicity at 25–28 °C and hydrophilicity at 40 °C. Surface hydrophobicity of the isolate was susceptible to various proteases (trypsin, pepsin, proteinase k and a-chymotrypsin) and inhibitors (SDS, mercaptoethanol and Triton X-100) and a significant reduction in CSH was recorded in hydrophobic conidia. Hydrophilic conidia remained more or less unaffected by such treatments. SDS-PAGE analysis of the hydrophobic and hydrophilic conidia exhibited several protein bands in the 25 to 61 kDa range. However, each protein population contained one protein that was not observed in the other population. For hydrophobic conidia, the unique protein had an apparent molecular mass of 49 kDa, while the unique protein associated with hydrophilic conidia had a molecular mass of 61 kDa. Our findings suggest that CSH and CSEC of mycoparasitic Trichoderma may contribute to non-specific adhesion on to the sclerotial surfaces of Macrophomina phaseolina that may be influenced by growth and environmental conditions.     

Taxonomy and secondary metabolites of <Emphasis Type="Italic">Pseudobotrytis</Emphasis> sp. FKA-25

Fungal strain FKA-25, isolated from forest soil collected on Yakushima Island, Kagoshima Prefecture, Japan, was assigned to genus Pseudobotrytis based on its morphological characteristics. Conidiophores were erect, slightly swollen at the end of the tip, and gave rise to umbellate conidiogenous cells that were in an expanded denticulate portion at the end and formed ellipsoidal to clavate conidia in sympodial succession. Identification as species P. terrestris was made on the basis of the character of 1-septate conidium. Although no secondary metabolites have been reported from the genus Pseudobotrytis, four secondary metabolite compounds (designated A to D) were isolated from the culture broth of strain FKA-25. Compounds B to D have been reported previously as FK-17-p2a, lunatinin, and 3,4-dihydro-3,4,8-trihydroxy-1(2H)-naphthalenone, respectively. Compound A was designated sespendole and possessed a novel indole-sesquiterpene skeleton.     

Bambusicolous fungi in Japan (8): a new species of <Emphasis Type="Italic">Pseudolachnella from</Emphasis> Yakushima Island,southern Japan

A coelomycetous fungus occurring on culms of Pleioblastus sp. in Yakushima Island, southern Japan, is described and illustrated as a new species, Pseudolachnella yakushimensis. The species is characterized by cupulate, superficial black setose conidiomata, and cylindrical 3-septate conidia with two to five appendages at each end. Pseudolachnella yakushimensis is similar to P. indica and P. scolecospora by its 3-septate conidia, but is different from them by its smaller conidia with more than two appendages.     

Low occurrence of patulin‐ and citrinin‐producing species isolated from grapes

Aims: To assess the ability of fungi isolated from grapes to produce patulin and citrinin. Methods and Results: A total of 446 Aspergillus isolates belonging to 20 species and 101 Penicillium isolates were inoculated in Czapek yeast extract agar and yeast extract sucrose agar and incubated for 7 days at 25°C. Extracts were analysed for patulin and citrinin by thin‐layer chromatography. None of the isolates of Aspergillus spp. produced either patulin or citrinin. Patulin was produced by three isolates of Penicillium expansum and two of Penicillium griseofulvum. Citrinin was produced by five isolates of P. expansum, two of Penicillium citrinum and one of Penicillium verrucosum. Conclusions: Our results show that the Aspergillus and Penicillium species commonly isolated from grapes are not a source of the mycotoxins, patulin and citrinin. Significance and Impact of the Study: The possibility of co‐occurrence of patulin and citrinin with ochratoxin A in grapes and grape products remain low, owing to the low frequency of isolation of potentially producing species.     

Scanning electron microscopy of Penicillium conidia

The morphology of conidia in 211 species and 12 varieties belonging to the genus Penicillium Link ex Gray have been studied and compared.According to surface ornamentation, conidia have been classified into six groups: A, smooth-walled (7% of the species); B, delicately roughened (13%); C, warty (28%); D, echinate (10%); E, striate with low irregular ridges (36%); and F, striate with scarce high ridges or bars (6%). Whereas the first two groups are closely related in both shape and average size, a gradual reduction was observed in size and in the length/width (l/w) ratio in the remaining groups. Echinate conidia were globose, having the largest average size. Only four species produced conidia not surpassing 2 m in diameter. Maximum length observed was 8 m, and most elongated conidia had a l/w ratio of 3.5. Forty per cent of the species studied had globose conidia.Conidia of the monoverticillate species were generally smaller, more globose and frequently with ridges. In the Asymmetrica, the conidia were generally larger, and showed ridges in comparatively few species. Conidia of the Symmetrica, which were frequently striate with ridges, presented the most elongated forms. The largest average size was found in the conidia of the Polyverticillata which were generally warty. Finally, we have considered the variations in surface ornamentation of conidia during the evolution of the genus Penicillium and drawn attention to their possible relationship with certain habitats and ways of conidial dispersion.