Mortierella sugadairana,a new homothallic species related to the firstly described heterothallic species in the genus

A new species, Mortierella sugadairana, is described for a fungus forming homothallic zygospores with a club-shaped macrosuspensor and a microsuspensor originating from the macrosuspensor. The species was isolated from cool regions in Japan and morphologically and phylogenetically close to a heterothallic species M. parvispora, which is the first species reported as a heterothallic species in the genus. Mycelial growth of the species was limited at 30 °C, whereas two isolates of M. parvispora can grow. This may indicate that the species and M. parvispora adapted to different climates from a common ancestor involving differentiation of the manner of reproduction.     

Bacterial chitin utilization at halophilic conditions

Chitin is a dominant structural polymer produced in large amounts by brine shrimp Artemia in hypersaline lakes. Microbiological analysis of chitin utilization as a growth substrate in hypersaline chloride–sulfate lakes in the south Kulunda Steppe (Altai, Russia) revealed two groups of bacteria able to grow on chitin at moderate salinity. Under aerobic conditions, an enrichment culture was obtained at 2 M NaCl. Further purification resulted in the isolation of strains HCh1 and strain HCh2, identified as representatives of the genera Saccharospirillum and Arhodomonas (both in the Gammaproteobacteria). The chitin-utilizing potential has not been previously recognized in these genera. The Saccharospirillum sp. strain HCh1 grew on chitin within the salinity range from 0.5 to 3.25 M NaCl (optimum at 1 M), while Arhodomonas sp. strain HCh2 grew up to 2.5 M NaCl but had a higher salt optimum at 1.5 M. Anaerobic enrichments grew with chitin at 2 and 4 M NaCl, but growth in the latter was extremely slow and the culture eventually lost viability. The enrichment at 2 M NaCl resulted in the isolation of strain HCh-An1, identified as a distant new species of the genus Orenia in the clostridial order Halanaerobiales. It was able to grow on chitin within a salinity range from 1.0 to 2.5 M NaCl (optimum at 1.5 M). The strain is proposed as a new species of the genus Orenia—O. chitinitropha.     

Nuclear DNA content variation of three Miscanthus species in China

In order to estimate the variation in nuclear genome size in Miscanthus, flow cytometry of nuclei stained by propidium iodide was carried out using 36 populations of three Miscanthus species: M. lutarioriparius, M. sacchariflorus and M. sinensis, which were sampled from cold northern to warm and humid southern and central China, as well as near the sea level in eastern China to mountains in western China. The DNA content of diploid was 4.37 ± 0.02 pg/2C in M. lutarioriparius, 4.37 ± 0.01 pg/2C in M. sacchariflorus, and 5.37 ± 0.03 pg/2C in M. sinensis, respectively. There was no intraspecific variation in the three Miscanthus species at the diploid level, suggesting that the genome size was stable within species and the diverse environments did not induce variation in genome size at the diploid level. However, tetraploid populations were found in M. lutarioriparius and M. sacchariflorus, and their genome sizes were 8.56 and 8.54 pg, respectively, which are lower than expected values (8.74 pg), indicating the genome downsizing after polyploidization in the genus. Our results showed that the plant height of M. lutarioriparius was the highest one among the three species and the species was more closely related to M. sacchariflorus than M. sinensis. The intra-species genomic variation and inter-species differentiation in Miscanthus species provide important genetic and genomic information for the development of Miscanthus, especially for the endemic species, M. lutarioriparius, (together with Miscanthus × giganteus) which are now emerging as a key bio-energy crop because of their high yields and strong adaptability.     

Species of the Metarhizium anisopliae complex with diverse ecological niches display different susceptibilities to antifungal agents

Species of the Metarhizium anisopliae complex are globally ubiquitous soil-inhabiting and predominantly insect-pathogenic fungi. The Metarhizium genus contains species ranging from specialists, such as Metarhizium acridum that only infects acridids, to generalists, such as M. anisopliae, Metarhizium brunneum, and Metarhizium robertsii that infect a broad range of insects and can also colonize plant roots. There is little information available about the susceptibility of Metarhizium species to clinical and non-clinical antifungal agents. We determined the susceptibility of 16 isolates comprising four Metarhizium species with different ecological niches to seven clinical (amphotericin B, ciclopirox olamine, fluconazole, griseofulvin, itraconazole, tebinafine, and voriconazole) and one non-clinical (benomyl) antifungal agents. All isolates of the specialist M. acridum were clearly more susceptible to most antifungals than the isolates of the generalists M. anisopliae sensu lato, M. brunneum, and M. robertsii. All isolates of M. anisopliae, M. brunneum, and M. robertsii were resistant to fluconazole and some were also resistant to amphotericin B. The marked differences in susceptibility between the specialist M. acridum and the generalist Metarhizium species suggest that this characteristic is associated with their different ecological niches, and may assist in devising rational antifungal treatments for the rare cases of mycoses caused by Metarhizium species.     

Responses of entomopathogenic fungi to the mutagen 4-nitroquinoline 1-oxide

Survival of entomopathogenic fungi under solar ultraviolet (UV) radiation is paramount to the success of biological control of insect pests and disease vectors. The mutagenic compound 4-nitroquinoline 1-oxide (4-NQO) is often used to mimic the biological effects of UV radiation on organisms. Therefore, we asked whether tolerance to 4-NQO could predict tolerance to UV radiation in thirty isolates of entomopathogenic fungi and one isolate of a xerophilic fungus. A dendrogram obtained from cluster analyses based on the 50 and 90 % inhibitory concentrations (IC₅₀ and IC₉₀, respectively) divided the fungal isolates into six clusters numbered consecutively based on their tolerance to 4-NQO. Cluster 6 contained species with highest tolerance to 4-NQO (IC₅₀ > 4.7 μM), including Mariannaea pruinosa, Lecanicillium aphanocladii, and Torrubiella homopterorum. Cluster 1 contained species least tolerant to 4-NQO (IC₅₀ < 0.2 μM), such as Metarhizium acridum (ARSEF 324), Tolypocladium geodes, and Metarhizium brunneum (ARSEF 7711). With few exceptions, the majority of Metarhizium species showed moderate to low tolerances (IC₅₀ between 0.4 and 0.9 μM) and were placed in cluster 2. Cluster 3 included species with moderate tolerance (IC₅₀ between 1.0 and 1.2 μM). In cluster 4 were species with moderate to high tolerance (IC₅₀ between 1.3 and 1.6 μM). Cluster 5 contained the species with high tolerance (IC₅₀ between 1.9 and 4.0 μM). The most UV tolerant isolate of M. acridum, ARSEF 324, was the least tolerant to 4-NQO. Also, L. aphanocladii, which is very susceptible to UV radiation, showed high tolerance to 4-NQO. Our results indicate that tolerance to 4-NQO does not correlate with tolerance to UV radiation. Therefore this chemical compound is not a predictor of UV tolerance in entomopathogenic fungi.     

The Tracing of Mycobacteria in Drinking Water Supply Systems by Culture, Conventional, and Real Time PCRs

Mycobacteria are widely present in diverse aquatic habitats, where they can survive for months or years while some species can even proliferate. The resistance of different mycobacterial species to disinfection methods like chlorination or ozonation could result in their presence in the final tap water of consumers. In this study, the culture method, Mycobacterium tuberculosis complex conventional duplex PCR for detection of non-tuberculous mycobacteria (NTM) and quantitative real-time PCR (qPCR) to detect three subspecies of M. avium species (M. a. avium, M. a. hominissuis, and M. a. paratuberculosis) were used to trace their possible path of transmission from the watershed through the reservoir and drinking water plant to raw drinking water and finally to households. A total of 124 samples from four drinking water supply systems in the Czech Republic, 52 dam sediments, 34 water treatment plant sludge samples, and 38 tap water household sediments, were analyzed. NTM of 11 different species were isolated by culture from 42 (33.9 %) samples; the most prevalent were M. gordonae (16.7 %), M. triplex (14.3 %), M. lentiflavum (9.5 %), M. a. avium (7.1 %), M. montefiorenase (7.1 %), and M. nonchromogenicum (7.1 %). NTM DNA was detected in 92 (76.7 %) samples. By qPCR analysis a statistically significant decrease (P < 0.01) was observed along the route from the reservoir (dam sediments), through water treatment sludge and finally to household sediments. The concentrations ranged from 10⁰ to 10⁴ DNA cells/g. It was confirmed that drinking water supply systems (watershed–reservoir–drinking water treatment plant–household) might be a potential transmission route for mycobacteria.     

Plant regeneration from the embryogenic calli of five major Miscanthus species, the non-food biomass crops

With the growing shortage of oil, coal, and other traditional fossil fuels, scientists in various fields have been looking for new fuel sources to solve the energy crisis. The genus Miscanthus is an ideal biofuel crop due to its rapid vegetative growth and its potential for high biomass yields. Plant regeneration through somatic embryogenesis is a viable method to achieve large-scale production of plant biomass. Callus induction from immature inflorescences of five Miscanthus species was performed on two different media, and the relative rates of callus proliferation were calculated. The highest multiplication coefficient, 3.92, was obtained with M. sacchariflorus ssp. lutarioriparius when the concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) in the medium was 4.0 mg/L, and this species also performed the best at the induction phase. The proliferation coefficient for M. floridulus increased to 3.19 when the concentration of 2,4-D was decreased from 4.0 to 2.0 mg/L. When the concentration of 2,4-D was 2.0 mg/L, the proliferation coefficient for M. sinensis was 2.47. In M. sacchariflorus, the proliferation coefficients were 2.89 and 2.49 for the lower and higher concentrations of 2,4-D, respectively. The multiplication coefficient of M. x giganteus was 2.60 on medium containing 4.0 mg/L 2,4-D. Three different regeneration media were tested to induce shoots in vitro. M. floridulus and M. sacchariflorus regenerated shoots at 100% frequency in three different regeneration media. The regeneration rate for M. sacchariflorus ssp. lutarioriparius reached 99.0% on medium containing 4.0 mg/L?N ⁶ -Benzylaminopurine (6-BA) and 0.1 mg/L α-Naphthaleneacetic acid (NAA). The best regeneration rate of M. sinensis was 35.2% using 2.0 mg/L 6-BA and 0.3 mg/L NAA, whereas the regeneration rate of M. x giganteus was 57.4% on medium supplemented with 3.0 mg/L 6-BA and 0.2 mg/L NAA. The in vitro-derived plantlets of all five species had 100% rooting rates on basal MS medium without supplementation. The survival rates of plantlets were above 90% for each species when subsequently grown outdoors.     

Separation of two Neotropical species: Macrothrix superaculeata (Smirnov, 1982) versus M. elegans Sars, 1901 (Macrothricidae, Anomopoda, Cladocera)

The morphology of two Neotropical taxa, Macrothrix elegans Sars, 1901 and M. superaculeata (Smirnov, 1982) (Macrothricidae, Anomopoda, Cladocera) was redescribed, based on type materials (a lectotype of M. elegans was selected here), and additional samples from the Americas. Previous conclusion about synonymy of both species was erroneous, because it was based on limited material from South America. M. superaculeata differs from M. elegans in the presence of a sharp spine at postero-dorsal angle of valves; a more fine ring around dorsal head pore; thinner hexagonal reticulation of valves; the presence of setules on basal segment of postabdominal seta; armature of exopod on antenna II, and some features of thoracic limbs. Previously, the discriminative features of the two species were not formulated accurately, and it was a reason of several misidentifications. Actually, M. superaculeata is found only in a limited set of localities from the Amazon basin, while M. elegans is one of the most common anomopod species in all Neotropics, from Argentina to Mexico.     

Mrakia arctica sp. nov., a new psychrophilic yeast isolated from an ice island in the Canadian High Arctic

Two strains of a psychrophilic basidiomycetous yeast species belonging to the genus Mrakia were isolated from a melt-pool mat community, on an ice island located in Disraeli Fjord, Ellesmere Island in the Canadian Arctic. Analysis of the large subunit rDNA D1/D2 domain and internal transcribed spacer region sequences indicated that these strains represent a novel species, and the name Mrakia arctica sp. nov. is proposed. This new species could grow at sub-zero temperatures and in vitamin-free media. Moreover, lipase and cellulase enzymes of M. arctica were strongly active even at ?3 °C. These results suggest an important role for M. arctica in the biogeochemical cycle of glacial ecosystems.     

Miscanthus: Genetic Diversity and Genotype Identification Using ISSR and RAPD Markers

Due to the limited number of molecular studies focused on European gene pool investigation, it is necessary to perform plant material recognition. Eighteen accessions of three Miscanthus species, namely, M. × giganteus, M. sinensis, M. sacchariflorus were evaluated with the use of molecular marker systems such as: inter simple sequence repeats (ISSRs), random amplified polymorphic DNA (RAPD), and by estimation of ploidy level based on flow cytometry. As a result, only one ISSR primer (ISSR1) and three RAPD primers (RAPD1, RAPD2, RAPD4) were required to identify all genotypes. Moreover, the use of the above mentioned molecular markers enable the proper species recognition of the interspecific hybrid M. × giganteus “Floridulus,” which has been previously mislabeled as M. floridulus. The highest genetic similarity coefficient (0.94) was observed between M. × giganteus clones, which indicates that the genetic diversity within this species was very low. Whereas M. sinensis genotypes represented a relatively wide diversity with similarity coefficient of 0.58. Cluster analysis using UPGMA grouped the 18 accessions in three clusters according to species affiliation including relabeled M. × giganteus “Floridulus,” which proved to be closely related to M.  × giganteus. Similar groupings were evident in the PCoA analysis.     

Feeding interactions between the introduced ctenophore Mnemiopsis leidyi and juvenile herring Clupea harengus in the Wadden Sea

We analysed feeding interactions between Mnemiopsis leidyi and juvenile Clupea harengus in the Wadden Sea. Biomass, diet overlap, prey selectivity, predation impact and stable isotope composition (C, N) of both species were assessed from June to September 2010. High biomass of C. harengus was found in June and July (wet weight 3.0 ± 1.8 g m^?3) followed by a steep decline from August to September (wet weight 0.01 ± 0.01 g m^?3), coinciding with a dramatic increase in M. leidyi biomass (wet weight 18.3 ± 16.1 g m^?3 during August). These two species showed a high overlap in their respective diets (copepods, meroplankton) during the study period. Predation impact of C. harengus on calanoid copepods was highest in June and July where 84 and 41 % of the standing stock were eaten per day in June and July, respectively. Predation impact of M. leidyi on calanoid copepods was highest in September (16 %). Based on stable isotope analysis C. harengus and M. leidyi were assigned to a trophic level of 3.08 and 2.47, respectively. Furthermore, we assessed the potential of competition between M. leidyi and C. harengus in a mesocosm experiment. Results indicated that at present zooplankton densities intraspecific competition in C. harengus seemed to be greater than interspecies competition with M. leidyi. Due to the low predation impact of M. leidyi and the reduced temporal overlap, competition between M. leidyi and C. harengus during the study period was estimated as low. Nevertheless, considering the high dietary overlap and the inter-annual variation in biomass and occurrence of both species and their zooplankton prey, competition in the Wadden Sea area cannot be excluded.     

Morpho-Molecular Characterization of Soil Inhabitant Dermatophytes from Ahvaz,Southwest of Iran,a High Occurrence of <Emphasis Type="Italic">Microsporum fulvum</Emphasis>

Occurrence and diversity of dermatophyte mycoflora in 298 soil samples from Ahvaz, Southwest of Iran was investigated by using the hair-baiting technique. The samples were collected during spring (n = 210) and autumn (n = 88) of 2015, and the fungal isolates were identified based on the macro- and micro-morphology of colonies and with further ITS-rDNA RFLP and sequencing. Totally, 60 soil samples (20.1%) were positive for dermatophyte growth whose pH varied from 7.0 to 7.9. The highest (26.6%) and the lowest (14.3%) recovery rates were from the animal resorts and the streets soils samples, respectively. Seasonally, 16.7% of the spring samples and 28.4% of the autumn samples were positive. Based on molecular identification, three species of two genera were identified viz. M. fulvum (n = 57), M. canis (n = 2) and zoophilic Trichophyton interdigitale (n = 1). As a specific goal in the study, differentiation of the species in Microsporum gypseum complex was established by measuring the mean length and width of macroconidia in some strains of M. gypseum, M. fulvum and M. incurvatum. Mean size for macroconidia length and width in three species showed that M. gypseum and M. incurvatum can morphologically be differentiated from M. fulvum but not from each other. M. fulvum was the most abundant species isolated from the soils of Ahvaz; however, to comprehensively specify the distribution pattern of geophilic dermatophytes in the soils of this city further investigations are needed. Identification based on micro-morphometric is not effective for species distinction in M. gypseum complex, while molecular procedures based on sequencing of certain DNA regions are the most reliable and applicable strategies for this purpose.     

Cloning,expression and characterization of a novel cold-active and halophilic xylanase from Zunongwangia profunda

A new xylanase gene (xynA) from the marine microorganism Zunongwangia profunda was identified to encode 374 amino acid residues. Its product (XynA) showed the highest identity (42.78 %) with a xylanase from Bacillus sp. SN5 among the characterized xylanases. XynA exhibited the highest activity at pH 6.5 and 30 °C, retaining 23 and 38 % of the optimal activity at 0 and 5 °C, respectively. XynA was not only cold active, but also halophilic, and both its activity and thermostability could be significantly increased by NaCl, showing the highest activity (180 % of the activity) at 3 M NaCl and retaining nearly 100 % activity at 5 M NaCl, compared to the absence of NaCl. In the presence of 3 M NaCl, the k _cat/K _m value of XynA exhibited a 3.41-fold increase for beechwood xylan compared to no added NaCl, and the residual activity of XynA increased from 23 % (no added NaCl) to 58 % after 1 h incubation at 45 °C. This may be the first report concerning a cold-adapted xylanase from a non-halophilic species that displays the highest activity at a NaCl concentration range from 3 to 5 M. The features of cold activity and salt tolerance suggest the potential application of XynA in the food industry and bioethanol production from marine seaweeds.     

Isolation and Identification of Environmental Mycobacteria in the Waters of a Hemodialysis Center

The use of poorly treated water during hemodialysis may lead to contamination with nontuberculous mycobacteria (NTM). This study aimed to isolate and identify NTM species in the water of a Brazilian hemodialysis center. We collected 210 samples of water from the hydric system of the unit (post-osmosis system, hemodialysis rooms, reuse system, and hemodialysis equipment) and from the municipal supply network; we isolated the NTM by a classic microbiological technique and identified them by the PCR restriction enzyme pattern of the hsp65 gene (PRA). Fifty-one (24.3 %) of the collected samples tested positive for NTM; both the municipal supply network (2 samples, 3.2 %) and the hydric system of the hemodialysis center (49 samples, 96.1 %) contained NTM. We isolated and identified potentially pathogenic bacteria such as Mycobacterium lentiflavum (59.0 %) and M. kansasii (5.0 %), as well as rarely pathogenic bacteria like M. gordonae (24.0 %), M. gastri (8.0 %), and M. szulgai (4.0 %). The ability of NTM to cause diseases is well documented in the literature. Therefore, the identification of NTM in the water of a Brazilian hemodialysis center calls for more effective water disinfection procedures in this unit.     

Clash of the titans: a multi-species invasion with high gene flow in the globally invasive titan acorn barnacle

The goal of this study was to investigate the phylogeny, invasion history and genetic structure of the global invader Megabalanus coccopoma. First, we created a Bayesian phylogeny using cytochrome oxidase I and 16S mitochondrial genes of samples we collected and sequences available on GenBank for all species in the genus Megabalanus. Second, we compared the genetic differences within and between native and invasive populations verified as M. coccopoma by constructing a haplotype network of the COI sequences and estimating gene diversity (h) and nucleotide diversity (π). Finally, we ran an analysis of molecular variance and calculated pairwise Φ _ST to evaluate the similarity among populations. We identified several lineages that correspond to putatively different species of Megabalanus and uncovered nomenclature discrepancies among GenBank samples and undocumented lineages from our own collections. However, we found that the majority of samples were indeed M. coccopoma. Among populations of M. coccopoma, levels of within-population genetic diversity were not significantly different (p _h  = 0.131, p _π  = 0.129) between native (h = 0.970, π = 0.00708) and non-native populations (h = 0.950, π = 0.00605) and analysis of molecular variance analyses revealed that 98.34 % of the genetic variation was partitioned within populations with a significant global Φ _ST  = 0.017. Our results revealed that invasions in at least the southeastern United States and Brazil are composed of multiple lineages; however, we found that most of the global invasion occurred from a single lineage, M. coccopoma, and that no significant genetic differentiation exists between native and non-native populations of this species.     

Comparative studies on phosphate utilization of two bloom-forming Microcystis spp. (cyanobacteria) isolated from Lake Taihu (China)

Microcystis is a common freshwater bloom-dominating cyanobacterial genus. However, the crucial factors that affect the seasonal succession and extent of dominance of different species remain largely unknown. This study investigated inorganic phosphorus (P) uptake, growth, and P utilization of two dominant Microcystis species from Lake Taihu (China) at different P concentrations and temperatures. Compared with Microcystis wesenbergii, Microcystis flos-aquae had higher maximum uptake rate and luxury storage coefficient. However, M. flos-aquae also had a higher P demand for growth. Under P-rich conditions (64.5 to 174.5 μM), M. flos-aquae had a higher growth rate (0.16 to 0.21 d^?1) than M. wesenbergii (0.06 to 0.19 d^?1) at 22 to 30 °C. M. wesenbergii, with higher phosphate affinity, had a lower P demand to sustain its growth, and yielded a higher growth rate of 0.10 d^?1 at low P concentration (6.5 μM) at 34 °C, whereas M. flos-aquae had a lower growth rate of 0.03 d^?1. Therefore, M. flos-aquae was dominant in late spring under P-rich conditions, whereas M. wesenbergii prevailed in hot summer after M. flos-aquae decreased under P-limited conditions. These results agree well with their succession patterns in the field. Overall, succession and dominance of M. flos-aquae and M. wesenbergii in Lake Taihu are regulated by P concentration and water temperature.     

A new thermophilic species of Myceliophthora from China

Myceliophthora guttulata sp. nov. is described and illustrated based on strains isolated from soil in China. This species is thermophilic with optimal growth temperature of 40–45 °C, and minimum growth temperature of 25 °C. Morphologically, this species is characterized by smooth, guttulate, pyriform to obovoid blastoconidia born directly on the side of hyphae, on long or short pedicels or in groups of 1–4 on ampulliform swellings. Phylogenetic analysis based on multi-locus alignment of internal transcribed spacer (ITS), elongation factor 1-alpha (EF1-α) and RNA polymerase II subunit (RPB2) regions showed that M. guttulata clustered within the genus Myceliophthora, and is closely related to four thermophilic species, i.e. M. fergusii, M. thermophila, M. heterothallic, and M. hinnulea.     

Xylodon subflaviporus sp. nov. (Hymenochaetales,Basidiomycota) from East Asia

Xylodon subflaviporus (Hymenochaetales, Basidiomycota) is described as a new species collected from tropical and subtropical regions of East Asia. This new species is characterized by resupinate basidiocarps, a poroid hymenophore, pseudodimitic hyphal system, nodose-septate hyphae, four types of cystidia (capitate, acicular to cylindrical, subulate to ventricose, and apically-encrusted), and thin-walled, colorless, smooth and ellipsoid basidiospores. Bayesian, Maximum Likelihood and Maximum Parsimony algorithms were used to construct phylogenies inferred from the internal transcribed spacer (ITS) region of rDNA. Morphological and molecular studies, along with intercompatibility tests, confirmed the independent status of this species. Also, based on morphological and molecular evidence, six new combinations of Xylodon are proposed to accommodate species originally classified under Hyphodontia s.l., i.e. X. bubalinus, X. chinensis, X. mollissimus, X. nongravis, X. reticulatus and X. subtropicus. A key to known species of Hyphodontia s.l. with poroid, irpicoid or raduloid hymenophores is provided for the convenience in identification.