Taxonomic revision of Aspergillus section Clavati based on molecular, morphological and physiological data

Aspergillus section Clavati has been revised using morphology, secondary metabolites, physiological characters and DNA sequences. Phylogenetic analysis of β-tubulin, ITS and calmodulin sequence data indicated that Aspergillus section Clavati includes 6 species, A. clavatus (synonyms: A. apicalis, A. pallidus), A. giganteus, A. rhizopodus, A. longivesica, Neocarpenteles acanthosporus and A. clavatonanicus. Neocarpenteles acanthosporus is the only known teleomorph of this section. The sister genera to Neocarpenteles are Neosartorya and Dichotomomyces based on sequence data. Species in Neosartorya and Neocarpenteles have anamorphs with green conidia and share the production of tryptoquivalins, while Dichotomomyces was found to be able to produce gliotoxin, which is also produced by some Neosartorya species, and tryptoquivalines and tryptoquivalones produced by members of both section Clavati and Fumigati. All species in section Clavati are alkalitolerant and acidotolerant and they all have clavate conidial heads. Many species are coprophilic and produce the effective antibiotic patulin. Members of section Clavati also produce antafumicin, tryptoquivalines, cytochalasins, sarcins, dehydrocarolic acid and kotanins (orlandin, desmethylkotanin and kotanin) in species specific combinations. Another species previously assigned to section Clavati, A. ingratus is considered a synonym of Hemicarpenteles paradoxus, which is phylogenetically very distantly related to Neocarpenteles and section Clavati

Taxonomy of Penicillium section Citrina

Species of Penicillium section Citrina have a worldwide distribution and occur commonly in soils. The section is here delimited using a combination of phenotypic characters and sequences of the nuclear ribosomal RNA gene operon, including the internal transcribed spacer regions ITS1 and ITS2, the 5.8S nrDNA (ITS) and partial RPB2 sequences. Species assigned to section Citrina share the production of symmetrically biverticillate conidiophores, flask shaped phialides (7.0–9.0 μm long) and relatively small conidia (2.0–3.0 μm diam). Some species can produce greyish-brown coloured cleistothecia containing flanged ascospores. In the present study, more than 250 isolates presumably belonging to section Citrina were examined using a combined analysis of phenotypic and physiological characters, extrolite profiles and ITS, β-tubulin and/or calmodulin sequences. Section Citrina includes 39 species, and 17 of those are described here as new. The most important phenotypic characters for distinguishing species are growth rates and colony reverse colours on the agar media CYA, MEA and YES; shape, size and ornamentation of conidia and the production of sclerotia or cleistothecia. Temperature-growth profiles were made for all examined species and are a valuable character characters for species identification. Species centered around P. citrinum generally have a higher maximum growth temperature (33–36 °C) than species related to P. westlingii (27–33 °C). Extrolite patterns and partial calmodulin and β-tubulin sequences can be used for sequence based identification and resolved all species. In contrast, ITS sequences were less variable and only 55 % of the species could be unambiguously identified with this locus

The effect of culture preservation techniques on patulin and citrinin production by Penicillium expansum Link

Aims: To study the influence of culture preservation methods and culture conditions on the production of the mycotoxins patulin and citrinin by Penicillium expansum. Methods and results: Ten strains of Penicillium expansum were preserved using subculture and maintenance at 4 ºC, mineral oil, drying on silica gel and freeze-drying. Patulin and citrinin production was assessed on yeast extract sucrose agar (YES) and grape juice agar (GJ), using TLC before and after 0.5, 2–3, 6 and 12 months preservation. Citrinin was detected in all cultures for all preservation techniques on YES. The patulin profiles obtained differed with strain and culture media used. Conclusions: Citrinin production seems to be a stable character for the tested strains. There is a tendency for patulin detection with time apparently more consistent for silica gel storage and freeze-drying, especially when the strains are grown on GJ. Significance and Impact of the Study: Variability in the profiles of the mycotoxins tested seems to be more strain-specific than dependent on the preservation technique used

Discovery of Aspergillus frankstonensis sp. nov. during environmental sampling for animal and human fungal pathogens

Invasive fungal infections, IFI, due to species in Aspergillus section Fumigati, ASF, including the Aspergillus viridinutans species complex, AVSC, are increasingly reported in humans and cats. The risk of exposure to these medically important fungi in Australia is unknown. Air and soil was sampled from the domiciles of pet cats diagnosed with these IFI and from a nature reserve in Frankston, Victoria, where Aspergillus viridinutans sensu stricto was discovered in, . Of, ASF species isolated, were A. fumigatus sensu stricto, were AVSC, A. felis-clade and A. frankstonensis sp. nov., and, were other species, . Seven pathogenic ASF species known to cause disease in humans and animals, A. felis-clade, A. fischeri, A. thermomutatus, A. lentulus, A. laciniosus A. fumisynnematus, A. hiratsukae, comprised, of isolates overall. AVSC species were only isolated from Frankston soil where they were abundant, suggesting a particular ecological niche. Phylogenetic, morphological and metabolomic analyses of these isolates identified a new species, A. frankstonensis that is phylogenetically distinct from other AVSC species, heterothallic and produces a unique array of extrolites, including the UV spectrum characterized compounds DOLD, RAIMO and CALBO. Shared morphological and physiological characteristics with other AVSC species include slow sporulation, optimal growth at, ÊC, no growth at, ÊC, and viriditoxin production. Overall, the risk of environmental exposure to pathogenic species in ASF in Australia appears to be high, but there was no evidence of direct environmental exposure to AVSC species in areas where humans and cats cohabitate.Jessica J. Talbot, Jos Houbraken, Jens C. Frisvad, Robert A. Samson, Sarah E. Kidd, John Pitt, Sue Lindsay, Julia A. Beatty, Vanessa R. Barr

Secondary metabolite profiling, growth profiles and other tools for species recognition and important Aspergillus mycotoxins

Species in the genus Aspergillus have been classified primarily based on morphological features. Sequencing of house-hold genes has also been used in Aspergillus taxonomy and phylogeny, while extrolites and physiological features have been used less frequently. Three independent ways of classifying and identifying aspergilli appear to be applicable: Morphology combined with physiology and nutritional features, secondary metabolite profiling and DNA sequencing. These three ways of identifying Aspergillus species often point to the same species. This consensus approach can be used initially, but if consensus is achieved it is recommended to combine at least two of these independent ways of characterising aspergilli in a polyphasic taxonomy. The chemical combination of secondary metabolites and DNA sequence features has not been explored in taxonomy yet, however. Examples of these different taxonomic approaches will be given for Aspergillus section Nigri

New and revisited species in Aspergillus section Nigri

Four new species, Aspergillus eucalypticola, A. neoniger, A. fijiensis and A. indologenus are described and illustrated. Aspergillus eucalypticola was isolated from Eucalyptus leaf from Australia, and is related to A. tubingensis and A. costaricaensis, but could clearly be distinguished from them based on either β-tubulin or calmodulin sequence data. Aspergillus eucalypticola produced pyranonigrin A, funalenone, aurasperone B and other naphtho-γ-pyrones. Aspergillus neoniger is also a biseriate species isolated from desert sand in Namibia, and mangrove water in Venezuela, which produces aurasperone B and pyranonigrin A. Aspergillus fijiensis is a uniseriate species related to A. aculeatinus, and was isolated from soil in Fiji, and from Lactuca sativa in Indonesia. This species is able to grow at 37 °C, and produces asperparalines and okaramins. Aspergillus indologenus was isolated from soil, India. This species also belongs to the uniseriate group of black aspergilli, and was found to be related to, but clearly distinguishable from A. uvarum based on β-tubulin, calmodulin and ITS sequence data. Aspergillus indologenus produced the insecticidal compounds okaramins A, B, H, and two types of indol-alkaloids which have not been structure elucidated. Two other species, A. violaceofuscus and A. acidus, are revalidated based on molecular and extrolite data. Aspergillus violaceofuscus was found to be related to A. japonicus, and produced some of the same interesting indol-alkaloids as A. indologenus, and also produced several families of partially characterised extrolites that were also found in A. heteromorphus. Aspergillus acidus (previously known as A. foetidus var. pallidus and A. foetidus var. acidus) is also a valid species, while A. foetidus is a synonym of A. niger based on molecular and physiological data. Two other species described previously, A. coreanus and A. lacticoffeatus, were found to be colour mutants of A. acidus and A. niger, respectively. Methods which could be used to distinguish the two closely related and economically important species A. niger and A. awamori are also detailed. Although these species differ in their occurrence and several physiological means (elastase activities, abilities to utilise 2-deoxy-D-glucose as sole carbon source), our data indicate that only molecular approaches including sequence analysis of calmodulin or β-tubulin genes, AFLP analysis, UP-PCR analysis or mtDNA RFLP analysis can be used reliably to distinguish these sibling species. Aspergillus section Nigri now includes 26 taxa

Diversity in secondary metabolites including mycotoxins from strains of aspergillus section nigri isolated from raw cashew nuts from Benin, West Africa

Open access JournalIn a previous study, raw cashew kernels were assayed for the fungal contamination focusing on strains belonging to the genus Aspergillus and on aflatoxins producers. These samples showed high contamination with Aspergillus section Nigri species and absence of aflatoxins. To investigate the diversity of secondary metabolites, including mycotoxins, the species of A. section Nigri may produce and thus threaten to contaminate the raw cashew kernels, 150 strains were isolated from cashew samples and assayed for their production of secondary metabolites using liquid chromatography high resolution mass spectrometry (LC-HRMS). Seven species of black Aspergilli were isolated based on morphological and chemical identification: A. tubingensis (44%), A. niger (32%), A. brasiliensis (10%), A. carbonarius (8.7%), A. luchuensis (2.7%), A. aculeatus (2%) and A. aculeatinus (0.7%). From these, 45 metabolites and their isomers were identified. Aurasperone and pyranonigrin A, produced by all species excluding A. aculeatus and A. aculeatinus, were most prevalent and were encountered in 146 (97.3%) and 145 (95.7%) isolates, respectively. Three mycotoxins groups were detected: fumonisins (B2 and B4) (2.7%) ochratoxin A (13.3%), and secalonic acids (2%), indicating that these mycotoxins could occur in raw cashew nuts. Thirty strains of black Aspergilli were randomly sampled for verification of species identity based on sequences of β-tubulin and calmodulin genes. Among them, 27 isolates were positive to the primers used and 11 were identified as A. niger, 7 as A. tubingensis, 6 as A. carbonarius, 2 as A. luchuensis and 1 as A. welwitschiae confirming the species names as based on morphology and chemical features. These strains clustered in 5 clades in A. section Nigri. Chemical profile clustering also showed also 5 groups confirming the species specific metabolites production

A re-evaluation of Penicillium section Canescentia, including the description of five new species

A survey of Penicillium in the fynbos biome from South Africa resulted in the isolation of 61 species of which 29 were found to be new. In this study we focus on Penicillium section Canescentia, providing a phylogenetic re-evaluation based on the analysis of partial beta-tubulin (BenA), calmodulin (CaM) and RNA polymerase II second largest subunit (RPB2) sequence data. Based on phylogenies we show that five fynbos species are new and several previously assigned synonyms of P. canescens and P. janczewskii should be considered as distinct species. As such, we provide descriptions for the five new species and introduce the new name P. elizabethiae for the illegitimate P. echinatum. We also update the accepted species list and synonymies of section Canescentia species and provide a review of extrolites produced by these species.South African Biosystematics Initiative (SABI); Foundational Biodiversity Information Programme (FBIP) of the National Research Foundation (NRF); Future Leaders - African Independent Research fellowship programme (FLAIR); Novo Nordisk Foundation; Project INTARACT.https://www.persoonia.orgpm2022BiochemistryForestry and Agricultural Biotechnology Institute (FABI)GeneticsMicrobiology and Plant Patholog

Mycotoxin production from fungi isolated from grapes

Aims: In order to assess the potential for producing mycotoxins, fungi were isolated from wine producing grapes. Methods and Results: The isolates were identified and Penicillium expansum, the most well recognized mycotoxin producer, was analysed for mycotoxin production by TLC. Many of the strains produced patulin and/or citrinin, often depending on whether they were grown on a grape or yeast extract sucrose media. Conclusions: Citrinin was produced by all strains grown in the yeast extract sucrose medium, but only one strain (from 51) was able to produce this compound in grape juice medium. Patulin was produced in the yeast extract medium by 20 strains and in grape juice medium by 33 strains. Significance and Impact of the Study: The presence of mycotoxins in wine producing grapes is discussed. Grapes contamination with patulin seems not to contribute to wine contamination, and no ochratoxin producing fungi was identified.The British Council, Conselho de Reitores das Universidades Portuguesas (CRUP) - Windsor Programme - grant no 29/00