Secondary metabolites of <Emphasis Type="Italic">Penicillium</Emphasis> fungi isolated from permafrost deposits as chemotaxonomic markers

Species identification of slow-growing fungi of the genus Penicillium isolated from ancient permafrost deposits was performed using micro- and macromorphological characteristics as well as the composition of secondary metabolites. The strains producing clavine ergot alkaloids fumigaclavines A and B and festuclavine were assigned to the species P. palitans Westling 1911, whereas the strains forming ochratoxins A and B were identified as P. verrucosum Dierckx 1901.     

Monitoring of ochratoxin A and ochratoxin-producing fungi in traditional salami manufactured in Northern Italy

Fungi have a crucial role in the correct maturation of salami, but special attention should be addressed to the production of the nephrotoxic, immunotoxic, and carcinogenic mycotoxin ochratoxin A (OTA). In a monitoring study conducted in Northern Italy, OTA was detected by liquid chromatography coupled with mass spectrometry in 13 out 133 samples of traditional salami (9.8% of the total count). Mycological analysis of these samples yielded 247 fungal isolates which were identified to species level. The most frequent species were Penicillium nalgiovense, P. solitum, and P. chrysogenum. P. nordicum, an OTA-producing species commonly found in proteinaceous food, was not found in these samples. Three isolates were found to be Aspergillus westerdijkiae, an OTA-producing species. In order to check the results of the microbiological identification, 19 different strains of Aspergillus and 94 of Penicillium were tested for the presence of a sequence common to OTA-producing fungi by real-time PCR. None of the studied isolates, including the three A. westerdijkiae, possessed the otanpsPN target which is common to OTA-producing strains. Two out of three isolates of the A. westerdijkiae were also PCR-negative for the otanpsPN gene and did not produce OTA in culture. Conversely, this target sequence was amplified from the DNA purified from 14 salami casings including three casings harboring A. westerdijkiae. The amplification of sequences specific for OTA-producing strains performed on total genomic DNA extracted directly from salami casings provided a more suitable approach than PCR analysis of isolates from salami for the OTA-related otanpsPN gene to evaluate the risk of OTA contamination.     

Occurrence of the toxigenic fungi (producers of aflatoxins and ochratoxin A) in foodstuffs in the Czech Republic 1999–2000

The occurrence of toxigenic fungi producing aflatoxins and ochratoxin A in foodstuffs was studied in the Czech Republic. Twenty five commodities were collected at twelve collection places in the Czech Republic (300 food samples). The presence of potentially toxigenicAspergillus flavus was observed in 28% of the sampled foods (black pepper, caraway seeds, fruit tea, black tea, oat flakes, fine flour, rolled oat flakes and semolina) in the year 1999, and in 25% of the sampled foods (black pepper, black tea, fine flour) in the year 2000.A tamarii (aflatoxins producer) was found in 3 black pepper samples (25%) in both years. Aflatoxins were detected in black pepper and caraway seed samples in the year 1999 and in sweet red pepper in the year 2000.A parasiticus andA nomius were not isolated. Aspergillus section Nigri (potential producer of ochratoxin A) was detected in some foodstuffs. Ochratoxin A was detected in raisins.Penicillium verrucosum andA ochraceus were not isolated from foodstuffs.     

Investigation of stored wheat mycoflora,reporting the<Emphasis Type="Italic">Fusarium</Emphasis> cf.<Emphasis Type="Italic">langsethiae</Emphasis> in three provinces of Iran during 2007

Wheat is the most important cereal produced in Iran. A mycological survey was carried out for the first time, on the stored wheat samples in Tehran, East Azarbayejan and Mazandaran provinces in 2007. Exogenous and endogenous fungi, were isolated by the method of flotation with Malachite green agar (MGA 0.25) and Freeze blotter techniques respectively. In this study, 46 species belonging to 23 different genera were isolated.Cladosporium spp. (57.1–89.2%) andAlternaria spp. (82.4–100%) species were the predominant fungal species identified as endogenous mycoflora. The predominant exogenous fungi werePenicillium spp. (78.4–92.8%) andAspergillus spp. (71.4–85.7%) species.Fusarium proliferatum was the most prevalent species ofFusarium isolates.Aspergillus niger (39.4%) andAspergillus flavus (36.7%) were the predominantAspergillus species identified as exogenous mycoflora.Aspergillus flavus (26.6%) was the predominantAspergillus species identified as endogenous mycoflora. Flotation method with MGA 0.25 recommended for isolating of hyaline fungi from wheat cereals. In this study one isolate fromFusarium species was isolated on the basis of morphology and ribosomal internal transcribed spacer classified asFusarium langsethiae but on the basis of partial translation elongation factor-1alpha gene grouped withFusarium sporotrichioides. To our knowledge, this is the first report aboutF. cf.langsethiae in Iran and Asia.     

Some new taxa ofPotentilla L.

Five new species and two varieties from the sectionMultifidae are described from the Sino-Himalayan region. They are referred to the affinity ofP. multifida L. andP. griffithii Hook. f.; one of the new species is probably related toP. gerardiana Lindl. and one is intermediate between the species of the sectionsMultifidae andNiveae.     

Biochemical evidence bearing on the domestication of<Emphasis Type="Italic">Phaseolus</Emphasis> (Fabaceae) beans

The genusPhaseolus (Fabaceae) consists of some 50 species, all of which are distributed in the Americas. Four of these contain cultigens.P. vulgaris (common bean),P. lunatus (lima bean),P. acutifolius (tepary bean),P. coccineus subsp.coccineus (runner bean); andP. coccineus subsp.polyanthus (no English vernacular name). Biochemical markers—phaseolin seed storage protein and isozymes—have provided new evidence on the organization of the first three species. Domestication has possibly caused a strong reduction in genetic diversity inP. vulgaris andP. acutifolius. BothP. vulgaris andP. lunatus cultivars result from at least two independent domestications, in Mesoamerica and in the Andes. These two species consist of two gene pools, each of which includes wild ancestors and their respective cultivated descendants. Our findings suggest the need for additional emphasis on genetic conservation of wild ancestors and their use in breeding programs and for a comparison of inter-gene pool vs. intra-gene pool crosses in breeding programs.     

Morphology of Cell Wall Pore Channels in the Genus <Emphasis Type="Italic">Euastrum</Emphasis> Ralfs (Desmidiales)

The study of the cell wall structure was performed for ten species of Euastrum: E. ansatum (Ehrenb.) Ralfs, E. bidentatum Näg., E. binale (Turp.) Ehrenb. ex Ralfs, E. dubium Näg., E. elegans (Bréb.) Kütz. ex Ralfs, E. germanicum (Schmidle) W. Krieger, E. oblongum (Grev.) Ralfs ex Ralfs, E. pectinatum Bréb. ex Bréb. in Ralfs, E. validum West et G.S.West, E. verrucosum (Ehrenb.) ex Ralfs. The investigation of the cell wall ultrastructure has established for the first time that the pore canal in 6 of 10 species always has a coiled form to some degree. Three species (E. germanicum, E. pectinatum, and E. verrucosum) have both coiled and straight canals, and E. ansatum has only straight ones. A new type of pores (P7), typical only for representatives of the genus Euastrum, was also noted. In addition, a simple and effective method of preparing desmidium algal cells for investigation with transmission electron microscope has been developed.     

Morphological and molecular taxonomy of novel species <Emphasis Type="Italic">Pleurotheciaceae</Emphasis> from freshwater habitats in Yunnan,China

The diversity of lignicolous freshwater fungi of the Greater Mekong Subregion are currently being studied. In this paper, 18 fresh collections of Pleurotheciaceae from submerged wood in freshwater are characterized based on morphological characters and analyses of ITS, LSU, SSU and RPB2 sequence data. Six new species of Pleurotheciella (P. aquatica, P. fusiformis, P. guttulata, P. lunata, P. saprophytica, P. submersa), one new Phaeoisaria species (Ph. aquatica) and one new Pleurothecium species (Pl. aquaticum) are introduced based on their distinct morphology and evidence from molecular phylogeny. Pleurotheciella uniseptata, Phaeoisaria clematidis and Pleurothecium pulneyense are also redescribed and phylogenetic relationships assessed herein, these species are the first records for China.     

Metal concentrations and mycorrhizal status of plants colonizing copper mine tailings: potential for revegetation

A field survey of metal concentrations and mycorrhizal status of plants growing on copper mine tailings was conducted in Anhui Province, China. Available phosphorus and organic matter in the tailings were very low. High concentrations of Pb, Zn, As and Cd as well as Cu were observed on some sites. The dominant plants growing on mine tailings belonged to the families Gramineae and Compositae, and the most widely distributed plant species wereImperata cylindrica, Cynodon dactylon andPaspalum distichum. Coreopsis drummondii also grew well on the arid sites but not on wet sites. Very low or zero arbuscular mycorrhizal (AM) fungal colonization was observed in most of the plants, but extensive mycorrhizal colonization was recorded in the roots ofC. drummondii andC. dactylon. Metal concentrations in plant tissues indicated that /.cylindrica andP. distichum utilized avoidance mechanisms to survive at high metal concentrations. The investigation suggests that remediation and revegetation of heavy metal contaminated sites might be facilitated by selection of tolerant plant species. Isolation of tolerant AM fungi may also be warranted.     

<Emphasis Type="Italic">Phialemoniopsis endophytica</Emphasis> sp. nov., a new species of endophytic fungi from <Emphasis Type="Italic">Luffa cylindrica</Emphasis> in Henan,China

During a survey of endophytic fungi in the cucurbit plants collected from Henan, China, a new species, Phialemoniopsis endophytica was isolated from the lower stem of Luffa cylindrica. It differs from other Phialemoniopsis species by its cylindrical to flask-shaped phialides, falcate conidia with blunt ends, ostiolate pycnidium-like conidiomata without marginal setae and ellipsoidal chlamydospores. Multi-locus (ITS, LSU, ACT, and TUB) phylogenetic analysis confirmed that P. endophytica is distinct from other species. A synopsis of the morphological characters of the new species is provided.     

A global survey of <Emphasis Type="Italic">Puccinia</Emphasis>-rust on Cucurbitaceae

In this paper, all known autoecious Puccinia species (rust fungi, Uredinales) on Cucurbitaceae are described and most of them illustrated. A key and information on their host range and distribution is presented. Four species, Puccinia antennata, P. arbor-miraculensis, P. hieroglyphica, and P. rhytidioderma are proposed as new. P. momordicae and P. trochomeriae are recognised as valid species different from P. cephalandrae. Uredo melothriae is a new combination for Uromyces melothriae. P. cucumeris is a new report for Namibia, P. momordicae for Zimbabwe and P. vanderystii for Kenya. The species appear to be host specific with regard to host tribes, but P. citrulli and P. cucumeris may pass tribal boundaries. Most species are morphologically very similar and are characterised by a unique set of teliospore characters. They are therefore regarded as a natural group. The distribution of this group in semiarid habitats, mainly from southern Africa to India, is discussed.     

Diversity and enzyme activity of <Emphasis Type="Italic">Penicillium</Emphasis> species associated with macroalgae in Jeju Island

A total of 28 strains of 19 Penicillium species were isolated in a survey of extracellular enzyme-producing fungi from macroalgae along the coast of Jeju Island of Korea. Penicillium species were identified based on morphological and β-tubulin sequence analyses. In addition, the halo-tolerance and enzyme activity of all strains were evaluated. The diversity of Penicillium strains isolated from brown algae was higher than the diversity of strains isolated from green and red algae. The commonly isolated species were Penicillium antarcticum, P. bialowiezense, P. brevicompactum, P. crustosum, P. oxalicum, P. rubens, P. sumatrense, and P. terrigenum. While many strains showed endoglucanase, β-glucosidase, and protease activity, no alginase activity was detected. There was a positive correlation between halo-tolerance and endoglucanase activity within Penicillium species. Among 19 Penicillium species, three species–P. kongii, P. olsonii, and P. viticola–have not been previously recorded in Korea.     

On the taxonomy ofPuccinia sieversiae Arth. and some remarks on the phylogeny of the rusts onGeeae

Puccinia tatrensis Urban onGeum reptans in Czechoslovakia is recognized as a subspecies ofPuccinia sieversiae Arth.Puccinia waldsteiniae, P. gei andP. gei-parviflori appear to be the most ancient rusts known on the tribeGeeae and it is assumed that their common ancestral form was already present in the Cretaceous period.Puccinia sieversiae is more advanced and has its centre of origin and spread in Central Asia. Phylogenetically most advanced and at the same time most differentiated from each other are species ofPhragmidium, i.e.P. circumvallatum andP. miyabeanum.     

Investigation of Ochratoxin A biosynthetic genes in<Emphasis Type="Italic">Penicillium verrucosum</Emphasis> by DDRT-PCR experiments: differential expression of OTA genes

A defined minimal medium has been developed which is either restrictive or permissive for the production of ochratoxin A byPenicillium verrucosum simply by changing the nitrogen and carbon source. The combination of ammonium/glycerin promoted, whereas the combination nitrate/glucose repressed ochratoxin production.In parallel mutants ofP verrucosum have been constructed by restriction endonuclease mediated integration, which were not able to produce ochratoxin. Different types of transformants, which either produce no detectable amounts of ochratoxin A but ochratoxin α, strains which produce only ochratoxin B and strains which produces none of these secondary metabolites, have been observed.     

Biosystematic study of Berlandiera (Compositae)

Berlandiera is revised to includeB. subacaulis, B. pumila, B. texana, andB. lyrata with its varietieslyrata andmacrophylla. All species may be induced to interbreed readily, producing F₁ progeny which are vigorous but with reduced fertility. Sympatric species produce natural hybrids which are proposed here asB. Xhumilis (=B. pumila XB. subacaulis) andB. Xbetonicifolia (=B. pumila XB. texana). All taxa investigated hadn = 15 chromosomes, including new counts forB. subacaulis and all six F₁ hybrids.     

Comparison of two marine sponge-associated<Emphasis Type="Italic">Penicillium</Emphasis> strains DQ25 and SC10: differences in secondary metabolites and their bioactivities

Two strains ofPenicillium, DQ25 and SC10, isolated from marine spongeHaliclona angulata (Bowerbank) andHymeniacidon sp. respectively, were subjected to stationary cultivation under GYP medium for 30 days. The fermentation extracts were undergone bioactivities assays against human pathogens, phytopathogenic fungi and brine shrimp (Artemia salina). Bioassays-guided compounds isolation was performed by Silica gel columns and Sephadex LH-20 chromatography. Spectroscopic methods were used to structures elucidation of the compounds. Results showed the activities of secondary metabolites of strain DQ25 were generally stronger than that of strain SC10. Major bioactive molecules isolated from strain DQ25 were a 1,4-naphthoquinone derivative and an unidentified alkaloid. The two components were not isolated from the extract of strain SC10. ITS sequences revealed that these two species have the greatest similarity withPenicillium vinaceum andPenicillium granulatum respectively.     

Fungi from leaves of lotus (<Emphasis Type="Italic">Nelumbo nucifera</Emphasis>)

In spite of the self-cleaning property of its leaves called the lotus effect, leaves of lotus (Nelumbo nucifera) provide a habitat for an unknown fungal diversity. The aim of this study was to detect and identify fungi from leaves of N. nucifera, including ectophytic, parasitic and endophytic fungi, in Taiwan using different collection strategies, as well as morphological and diverse molecular markers established in the different systematic groups of fungi. Among ectophytic and parasitic fungi, a new species of Dissoconium and of Pseudocercospora are described, respectively. Phyllosticta nelumbonis Sawada is transferred to Diaporthe. Among plant parasitic fungi, Erysiphe takamatsui and Ps. nymphaeacea are recorded in Taiwan for the first time. Euryale is recorded as a new host genus for Ps. nymphaeacea. The basidiomycetous yeast Fereydounia khargensis is recorded for the first time from living plants and in East Asia. Endophytic fungi from lotus were studied for the first time. From 1002 plant segments, 476 endophytic isolates were produced in culture, comprising 33 typical terrestrial species mainly belonging to the genera Colletotrichum (mainly C. siamense), Diaporthe (D. tulliensis and D. ueckerae) and Fusarium (F. solani species 6, hitherto known from clinical samples), as well as to Xylariaceae, but no Ingoldian fungi. Most isolates were from leaf laminas (71%) compared to those from petioles (29%). From this observation, we conclude that the fungi of the aquatic lotus plant appear to have terrestrial origin and, after dispersal by wind and in spite of the lotus effect, may enter the plant from the lamina. Only three species isolated as endophytes were also found as ectophytic or parasitic fungi.     

Phylogenetic analysis of <Emphasis Type="Italic">Plectosphaerella</Emphasis> species based on multi-locus DNA sequences and description of <Emphasis Type="Italic">P. sinensis</Emphasis> sp. nov.

Species in Plectosphaerella are well known as pathogens of several plant species causing fruit, root and collar rot and collapse. In an investigation of endophytic fungi associated with cucurbit plants in China, we isolated 77 strains belonging to the genus Plectosphaerella. To identify the isolated strains, we collected the type or reference strains of all currently accepted species in Plectosphaerella except P. oratosquillae and conducted a phylogenetic analysis. Phylogenetic analysis of the partial 28S rDNA sequences showed that all species in Plectosphaerella were located in one clade of Plectosphaerellaceae. Based on multi-locus phylogenetic analysis of the ITS, CaM, EF1, TUB and morphological characteristics, all species in Plectosphaerella were well separated. Three endophytic strains from stems of Cucurbita moschata, Citrullus lanatus and Cucumis melo from North China were assigned to a new species described as P. sinensis in this paper. The new species differs morphologically from other Plectosphaerella species by irregular chlamydospores, and the dimensions of phialides and conidia. The other endophytic strains from several cucurbit plants were identified as P. cucumerina.