Haloalkalitolerant Fungi from Sediments of the Big Tambukan Saline Lake (Northern Caucasus): Diversity and Antimicrobial Potential.

We have performed a characterization of cultivated haloalkalitolerant fungi from the sediments of Big Tambukan Lake in order to assess their biodiversity and antimicrobial activity. This saline, slightly alkaline lake is known as a source of therapeutic sulfide mud used in sanatoria of the Caucasian Mineral Waters, Russia. Though data on bacteria and algae observed in this lake are available in the literature, data on fungi adapted to the conditions of the lake are lacking. The diversity of haloalkalitolerant fungi was low and represented by ascomycetes of the genera Acremonium, Alternaria, Aspergillus, Chordomyces, Emericellopsis, Fusarium, Gibellulopsis, Myriodontium, Penicillium, and Pseudeurotium. Most of the fungi were characterized by moderate alkaline resistance, and they tolerated NaCl concentrations up to 10% w/v. The analysis of the antimicrobial activity of fungi showed that 87.5% of all strains were active against Bacillus subtilis, and 39.6% were also determined to be effective against Escherichia coli. The majority of the strains were also active against Aspergillus niger and Candida albicans, about 66.7% and 62.5%, respectively. These studies indicate, for the first time, the presence of polyextremotolerant fungi in the sediments of Big Tambukan Lake, which probably reflects their involvement in the formation of therapeutic muds.

Trichocladium solani sp. nov.-A New Pathogen on Potato Tubers Causing Yellow Rot.

A new species, Trichocladium solani, was isolated from potato (Solanum tuberosum L.) tubers from Russia. The species has no observed teleomorph and is characterized morphologically by non-specific Acremonium-like conidia on single phialides and chains of swollen chlamydospores. Phylogenetic analysis placed the new species in a monophyletic clade inside the Trichocladium lineage with a high level of support from a multi-locus analysis of three gene regions: ITS, tub2, and rpb2. ITS is found to be insufficient for species delimitation and is not recommended for identification purposes in screening studies. T. solani is pathogenic to potato tubers and causes lesions that look similar to symptoms of Fusarium dry rot infection but with yellowish or greenish tint in the necrotized area. The disease has been named "yellow rot of potato tubers".

First report of Alternaria alternariacida causing potato leaf blight in the Far East, Russia.

Early blight of potato (Solanum tuberosum) is caused by Alternaria species and occurs annually in major potato producing regions of Russia. Diseased potato leaves displaying early blight symptoms were collected in July 2016 from a commercial field in Primorsky Krai, Russia (43.8242° N, 131.6219° E). The disease incidence was 30 to 40%. The initial symptoms appeared as typical early blight symptoms with a dark brown margin and diffused chlorosis on the leaf blade. Symptomatic leaves from different plants were randomly collected to isolate axenic cultures of the causal agents. Infected leaves were placed in wet chambers (moist filter papers in Petri dishes), and incubated at 25°C, 16 h/8 h dark/light photoperiod for 2-4 d. Single conidia were transferred to potato dextrose agar (PDA, Crous et al. 2009) in Petri dishes and incubated at 25°C for 7 d in the dark. Colonies were white-olivaceous, reverse side - olivaceous. Isolates were transferred onto potato carrot agar (PCA, Crous et al. 2009) and incubated at 22°C under a 16 h/8 h dark/light photoperiod for 7 d to stimulate sporulation. Most isolates (85%) were identified as A. protenta according to the morphological characteristics and molecular data, although one isolate showed sporulation that was somewhat atypical, having a smaller (especially narrower or more slender) conidia. Conidiophores were long, erect, and 65 to 100 µm × 5 to 6 µm in size. Conidia were solitary, long-ovoid in body with six to eight transverse septa, and 85 to 100 µm× 6 to 10 µm in size. Conidial beaks were filamentous, 110 to 200 µm × 2 to 5 µm in size. Genomic DNA was extracted from cultured isolates using the CTAB-chloroform extraction method (Griffith & Shaw 1998), and five gene regions including the internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (tef1), RNA polymerase second largest subunit (rpb2), Alternaria major allergen (Alt a 1) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified with the primer pairs ITS1/ITS4, EF1-728F/EF1-986R, RPB2-5F2/FRPB2-7cR, Alt-for/Alt-rev, and gpd1/gpd2 respectively (Woudenberg et al. 2014). PCR products were Sanger sequenced. All sequences for isolate A16PrPL21 were identical to isolate CBS 105.51; (accession nos.: ITS, KJ718105; tef1, KJ718454; rpb2, KJ718279; Alt a 1, KJ718625; gpd KJ717959) of A. alternariacida Woudenb. & Crous. ITS, tef1, rpb2, Alt a 1 and gpd sequences were deposited in GenBank under the accessions OM348531, MN580518, MN580529, MN562258 and MN544404 respectively. Based on morphological and molecular data, the isolate was identified as Alternaria alternariacida Woudenb. & Crous. A. alternariacida is closely related to A. silybi, which is also found in the Far East (Woudenberg et al. 2014). Phylogenetic distances between these strains are based on differences at the ITS, TEF1 and Alt a 1 gene regions. A pathogenicity test was carried out with isolate A16PrPL21 on nine 5-week-old healthy potato plants (cv. Nevsky) grown in a greenhouse at 23 ± 2 °C. Seed tubers were grown in the greenhouse to obtain the seedlings. Inoculation was performed by spraying a conidial suspension (105 spores/ml) prepared from 10-day-old A. alternariacida culture grown on PCA at 23° C with a 12-h photoperiod. Nine negative control plants were treated with sterile distilled water. The inoculated plants were then maintained in a greenhouse at 25 °C with high humidity and 12-h light period. All test plants were covered with plastic bags for 24 h to maintain high relative humidity and incubated at 24 to 28°C. Leaf spot symptoms, brown lesions with chlorotic halos, similar to those previously observed in naturally infected plants, appeared 5 d post inoculation for all inoculated plants. After 7 d, the spots reached 18 to 25 mm in size. The symptoms were similar to the original symptoms that occurred in the field. Negative control leaves were symptomless. Koch's postulates were fulfilled by reisolating the pathogen from inoculated leaves and identified as A. alternaricida by rpb2 gene sequence and morphological characteristics. To our knowledge, this is the first report of disease caused by A. alternaricida on potato plants. Early blight, caused by large-spored Alternaria, is a widespread disease on potato. A. alternaricida is among a group of species that cause early blight, according to current research. Studies of the Alternaria species' biology and analyses of their distribution are important for improving potato protection from early blight.

Isolation and Characterization of a Novel Hydrophobin, Sa-HFB1, with Antifungal Activity from an Alkaliphilic Fungus, Sodiomyces alkalinus.

The adaptations that alkaliphilic microorganisms have developed due to their extreme habitats promote the production of active natural compounds with the potential to control microorganisms, causing infections associated with healthcare. The primary purpose of this study was to isolate and identify a hydrophobin, Sa-HFB1, from an alkaliphilic fungus, Sodiomyces alkalinus. A potential antifungal effect against pathogenic and opportunistic fungi strains was determined. The MICs of Sa-HFB1 against opportunistic and clinical fungi ranged from 1 to 8 µg/mL and confirmed its higher activity against both non- and clinical isolates. The highest level of antifungal activity (MIC 1 µg/mL) was demonstrated for the clinical isolate Cryptococcus neoformans 297 m. The hydrophobin Sa-HFB1 may be partly responsible for the reported antifungal activity of S. alkalinus, and may serve as a potential source of lead compounds, meaning that it can be developed as an antifungal drug candidate.

Exploring Peptaibol's Profile, Antifungal, and Antitumor Activity of Emericellipsin A of Emericellopsis Species from Soda and Saline Soils.

Features of the biochemical adaptations of alkaliphilic fungi to exist in extreme environments could promote the production of active antibiotic compounds with the potential to control microorganisms, causing infections associated with health care. Thirty-eight alkaliphilic and alkalitolerant Emericellopsis strains (E. alkalina, E. cf. maritima, E. cf. terricola, Emericellopsis sp.) isolated from different saline soda soils and belonging to marine, terrestrial, and soda soil ecological clades were investigated for emericellipsin A (EmiA) biosynthesis, an antifungal peptaibol previously described for Emericellopsis alkalina. The analysis of the Emericellopsis sp. strains belonging to marine and terrestrial clades from chloride soils revealed another novel form with a mass of 1032.7 Da, defined by MALDI-TOF Ms/Ms spectrometers, as the EmiA lacked a hydroxyl (dEmiA). EmiA displayed strong inhibitory effects on cell proliferation and viability of HCT 116 cells in a dose- and time-dependent manners and induced apoptosis.

Significance of incongruent DNA loci in the taxonomy of wood-decaying Basidioradulum radula.

Modern taxonomic studies of Agaricomycetes rely on the integrative analyses of morphology, environmental data, geographic distribution, and usually several DNA loci. However, sampling and selection of DNA loci for the analyses are commonly shallow. In this study, we suggest minimal numbers of necessary specimens to sample and DNA loci to analyze in order to prevent inadequate taxonomic decisions in species groups with minor morphological and genealogical differences. We sampled four unlinked nuclear DNA gene regions (nuc rDNA ITS1-5.8S-ITS2, gh63, rpb2, and tef1) to revise the systematics of a common wood-decaying species Basidioradulum radula (Hymenochaetales) on an intercontinental set of specimens collected in the Northern Hemisphere. The DNA loci analyzed violate the genealogical concordance phylogenetic species recognition principles, thus confirming a single-species interpretation. We conclude that Hyphodontia syringae is a younger synonym of B. radula.