E. N. Bilanenko

On the diversity of fungi from soda soils

The diversity of filamentous fungi that can grow at high ambient pH values (i.e., 8–11) remains largely understudied. Here we study 100 alkalitolerant and alkaliphilic isolates from the soils around the basin of soda lakes in Asia and Africa to assess the major evolutionary lineages and morphologies pertinent to the alkaliphilic trait in filamentous fungi. The Emericellopsis lineage (Hypocreales, Hypocreomycetidae), along with Plectosphaerellaceae (Hypocreomycetidae), Pleosporaceae (Dothideomycetes), Chaetomiaceae (Sordariomycetidae) families appeared to be overrepresented with strong alkalitolerants and effective alkaliphiles. In particular, Sodiomyces species (Plectosphaerellaceae), Acrostalagmus luteoalbus (Plectosphaerellaceae), Emericellopsis alkalina (Hypocreales), Thielavia sp. (Chaetomiaceae), and Alternaria sect. Soda (Pleosporaceae) grew best at high ambient pH. The pH tolerance of Chordomyces antarcticum, Acrostalagmus luteoalbus and some other species was largely affected by the presence of extra Na+ in the growth medium. Moderate alkalitolerants included Scopulariopsis members (Microascales), Fusarium, Cladosporium, and many asexual acremonium-like species from Bionectriaceae. Weak alkalitolerants were represented by sporadic isolates of Penicillium, Purpureocillium lilacinum, and Alternaria alternata species, with the growth optimum at neutral or acidic pH. Weak alkalitolerants develop loose dry chains of spores easily dispersed by air. Their presence at low frequency with the growth optimum at neutral or acidic pH leads us to treat them as transient species in the alkaline soils, as those are also ubiquitous saprobes in normal soils. Our phylogenetic analyses show that the alkaliphilic trait in filamentous fungi has evolved several times. Several lineages harboring strong alkalitolerants derived from the known marine-borne fungi (Emericellopsis, Alternaria sect. Phragmosporae), or fall within the fungi associated with halophytic grasses (Pleosporaceae). Soda soils contain a diversity of fungi that range from weak alkalitolerant to alkaliphilic, which in few cases is associated with darkly pigmented mycelium and formation of microsclerotia. The alkaliphilic trait is spread throughout the Ascomycota, and usually juxtaposes with slime-covered polyphyletic acremonium-, verticillium-, gliocladium-types of asexual morphology, hyphae aggregating in chords, and enclosed fruit bodies.

Sodiomyces alkalinus, a new holomorphic alkaliphilic ascomycete within the Plectosphaerellaceae.

In this study we reassess the taxonomic reference of the previously described holomorphic alkaliphilic fungus Heleococcum alkalinum isolated from soda soils in Russia, Mongolia and Tanzania. We show that it is not an actual member of the genus Heleococcum (order Hypocreales) as stated before and should, therefore, be excluded from it and renamed. Multi-locus gene phylogeny analyses (based on nuclear ITS, 5.8S rDNA, 28S rDNA, 18S rDNA, RPB2 and TEF1-alpha) have displayed this fungus as a new taxon at the genus level within the family Plectosphaerellaceae, Hypocreomycetidae, Ascomycota. The reference species of actual Heleococcum members showed clear divergence from the strongly supported Heleococcum alkalinum position within the Plectosphaerellaceae, sister to the family Glomerellaceae. Eighteen strains isolated from soda lakes around the world show remarkable genetic similarity promoting speculations on their possible evolution in harsh alkaline environments. We established the pH growth optimum of this alkaliphilic fungus at c. pH 10 and tested growth on 30 carbon sources at pH 7 and 10. The new genus and species, Sodiomyces alkalinus gen. nov. comb. nov., is the second holomorphic fungus known within the family, the first one being Plectosphaerella – some members of this genus are known to be alkalitolerant. We propose the Plectosphaerellaceae family to be the source of alkaliphilic filamentous fungi as also the species known as Acremonium alcalophilum belongs to this group.

Are alkalitolerant fungi of the Emericellopsis lineage (Bionectriaceae) of marine origin

Surveying the fungi of alkaline soils in Siberia, Trans-Baikal regions (Russia), the Aral lake (Kazakhstan), and Eastern Mongolia, we report an abundance of alkalitolerant species representing the Emericellopsis-clade within the Acremonium cluster of fungi (order Hypocreales). On an alkaline medium (pH ca. 10), 34 acremonium-like fungal strains were obtained. One of these was able to develop a sexual morph and was shown to be a new member of the genus Emericellopsis, described here as E. alkalina sp. nov. Previous studies showed two distinct ecological clades within Emericellopsis, one consisting of terrestrial isolates and one predominantly marine. Remarkably, all the isolates from our study sites show high phylogenetic similarity based on six loci (LSU and SSU rDNA, RPB2, TEF1-α, β-tub and ITS region), regardless of their provenance within a broad geographical distribution. They group within the known marine-origin species, a finding that provides a possible link to the evolution of the alkaliphilic trait in the Emericellopsis lineage. We tested the capacities of all newly isolated strains, and the few available reference ex-type cultures, to grow over wide pH ranges. The growth performance varied among the tested isolates, which showed differences in growth rate as well as in pH preference. Whereas every newly isolated strain from soda soils was extremely alkalitolerant and displayed the ability to grow over a wide range of ambient pH (range 4–11.2), reference marine-borne and terrestrial strains showed moderate and no alkalitolerance, respectively. The growth pattern of the alkalitolerant Emericellopsis isolates was unlike that of the recently described and taxonomically unrelated alkaliphilic Sodiomyces alkalinus, obtained from the same type of soils but which showed a narrower preference towards high pH.

Effect of sodium chloride concentration in the medium on the composition of the membrane lipids and carbohydrates in the cytosol of the fungus Fusarium sp.

The fungus Fusarium sp. isolated from saline soil was identified by the ITS1-5.8S-ITS2 and the D1/D2 domains of LSU RNA as a member of the Fusarium incarnatum-equiseti species group. Its growth patterns on media with different NaCl concentrations indicated its adaptation as halotolerance. The mechanisms of halotolerance included accumulation of arabitol (a five-atom noncyclic polyol), a decreased sterols/phospholipids ratio, elevated level of phosphatidic acids in the phospholipids, and increased unsaturation of phospholipids, which was especially pronounced in the idiophase. The mechanisms of halotolerance of the mycelial fungus Fusarium sp. are discussed in comparison with yeasts and yeastlike fungi.

Effect of water activity and relative air humidity on the growth of Penicillium chrysogenum thom, Aspergillus repens (Corda) Sacc., and Trichoderma viride Pers. isolated from living spaces

Original data on the growth parameters of the fungi Penicillium chrysogenum Thom, Aspergillus repens (Corda) Sacc., and Trichoderma viride Pers. isolated from living spaces in Moscow are presented. Spore germination, fungal growth, and the radial growth rate of the colonies were investigated upon cultivation on agarized nutrient media with different water activity (aw) values. Spore germination and fungal growth were studied in house dust under laboratory conditions at different relative air humidity (RH). It was shown that, at decreased aw and RH, the spore germination time increased, as did the period from germination to mycelium and conidia formation, while the radial growth rate of colonies decreased. House dust was found to be a suitable growth substrate for A. repens and P. chrysogenum, supporting their complete life cycle. It was suggested that house dust is unsuitable as a substrate for the growth of T. viride. The aw and RH ranges for development of these micromycetes were determined. On this basis, the A. repens, P. chrysogenum, and T. viride strains isolated from living spaces were identified as xerophilic, xerotolerant, and hygrophilic ones, respectively.

The diversity of microfungi in peatlands originated from the White Sea

The diversity of culturable filamentous microfungi in peat and sediments of four peatlands at the coastal zone of Kandalaksha Bay of the White Sea (Murmansk region, Russia) was studied by culture methods on standard and selective media. Annually 100 samples were collected from the bogs 2007–2010. Based on morphological, molecular markers and cultural features, 211 taxa were identified. Fungal communities observed at the peatlands were influenced mostly by their sea origin. We discovered a large difference between fungal communities from the peat and the sediments of the peatlands. In contrast to the sediments, the fungal community of the peat was found to be consistent throughout sampling sites. Fungi with specific ecophysiology, such as Sphagnum-decomposing species (Oidiodendron griseum, O. tenuissimum. Penicillium spinulosum, P. thomii, Talaromyces funiculosus), psychrotolerant and associated with insects species (Pseudogymnoascus pannorum, Tolypocladium spp.), typical marine species (Acremonium spp.) were found. In addition, different types of sterile mycelia were characteristic for the researched peatlands.

Mycelial fungi in saline soils of the western Transbaikal region

Mycelial fungi in a solonchakous chestnut soil, a sulfate solonchak, and a soda solonchak were studied in the western Lake Baikal basin. The humus content, the degree of salinization, and the composition of salts affected the structure of the micromycete communities. In the chestnut soil, more than half of the species identified were found. The species diversity was higher in the nonsaline and humus horizons. The eolian deposit on the soda solonchak was characterized by the presence of six species of fungi that did not occur in other soil horizons. Their occurrence in the fresh deposit seems to be related to the eolian transfer of fungi with the fine earth from the adjacent areas with the nonsaline soils. The soda solonchak fungi are dominated by the haloalkalitolerant and alkalophilic micromycetes, among which the representatives of the Acremonium (A. antarcticum and A. rutilum) and Verticillum genera and Mycelia sterilia were identified. There was also an alkalophilic ascomycete, which is an indicator of soda salinization—Heleococcum alkalinum. On the whole, the soda solonchak had the lowest number of fungal germs and the lowest species diversity of mycelial fungi among the soils compared.

Communities of halotolerant micromycetes from the areas of natural salinity

The halophilic community of natural hypersaline soils has been isolated. Species of the genera Penicillium, Aspergillus, Cladosporium, as well as dark-colored yeast-like organisms, are most frequently isolated on selective media from all saline soil samples that we have studied. It has been shown that the community of micromycetes of hypersaline habitats is less dependent on geographical position than on the physicochemical parameters of habitat.

Alkalitolerant micromycetes in acidic and neutral soils of the temperate zone

A wide diversity of micromycetes from various taxonomic groups in acidic and neutral soils is known from the literature data. In the present work, the fungi isolated from these soils and capable of growth at high pH are analyzed. The fungi were isolated from acidic sod-podzol and neutral cultivated soils by plating on alkaline agar (pH 10.0–10.5). Their identification was carried out using morphological, cultural, and molecular genetic criteria. Phylogenetic analysis was performed and the rates of linear growth within a broad pH range (4.0–10.4) were determined. The isolates represented a polyphyletic group of ascomycetes (Sordariomycetes), which included members of Plectosphaerellaceae (5 species) and various families of Hypocreales (4 species). The most common species were Gibellulopsis nigrescens, Acrostalagmus luteoalbus, Chordomyces antarcticum, and Plectosphaerella spp. Investigation of fungal growth at different pH values revealed all isolates to be alkalitolerant, with no alkaliphilic fungi isolated from acidic sod-podzol and neutral cultivated soils. Although the group of isolates was polyphyletic and its members originated from different ecological and trophic niches, most alkalitolerant isolates exhibited common morphological traits with acremonium- and verticillium-like conidial spore formation, abundant slime formation, and a tendency for aggregation of their mycelium in bundles. Our research confirmed the presence of fungi with alkalitolerant adaptation to external pH in the sod-podzolic and cultivated soils of the Moscow region.

Characterization of the strain Monascus floridanus P. F. Cannon & E. L. Barnard, isolated from aviation fuel

A fungus was isolated from aviation fuel and identified as Monascus floridanus P.F. Cannon & E.L. Barnard (FR827895) according to its morphological and genetic properties. The isolate has some properties that are unusual for the type strain, including a prominent stripe on one of the sides of the ascospores and occurrence, along with the known Basipetospora-type thallic conidia, of the phialophora-like spore formation. The isolated strain Monascus floridanus, like the known kerosene fungus Hormoconis resinae (Lindau) Arx & G.A. de Vries, is capable of active growth in aviation fuel.