Nt Substitutions Research Articles

Bradyrhizobium japonicum HmuP is an RNA-binding protein that positively controls hmuR operon expression by suppression of a negative regulatory RNA element in the 5' untranslated region.

The hmuR operon encodes proteins for the uptake and utilization of heme as a nutritional iron source in Bradyrhizobium japonicum. The hmuR operon is transcriptionally activated by the Irr protein and is also positively controlled by HmuP by an unknown mechanism. An hmuP mutant does not express the hmuR operon genes nor does it grow on heme. Here, we show that hmuR expression from a heterologous promoter still requires hmuP, suggesting that HmuP does not regulate at the transcriptional level. Replacement of the 5' untranslated region (5'UTR) of an HmuP-independent gene with the hmuR 5'UTR conferred HmuP-dependent expression on that gene. Recombinant HmuP bound an HmuP-responsive RNA element (HPRE) within the hmuR 5'UTR. A 2 nt substitution predicted to destabilize the secondary structure of the HPRE abolished both HmuP binding activity invitro and hmuR expression in cells. However, deletion of the HPRE partially restored hmuR expression in an hmuP mutant, and it rescued growth of the hmuP mutant on heme. These findings suggest that the HPRE is a negative regulatory RNA element that is suppressed when bound by HmuP to express the hmuR operon.

A comprehensive molecular analysis of bovine coronavirus strains isolated from Brazil and comparison of a wild-type and cell culture-adapted strain associated with respiratory disease.

Bovine coronavirus (BCoV) has dual tropisms that can trigger enteric and respiratory diseases in cattle. Despite its global distribution, BCoV field strains from Brazil remain underexplored in studies investigating the virus's worldwide circulation. Another research gap involves the comparative analysis of S protein sequences in BCoV isolates from passages in cell lines versus direct sequencing from clinical samples. Therefore, one of the objectives of our study was to conduct a comprehensive phylogenetic analysis of BCoV strains identified from Brazil, including a respiratory strain obtained during this study, comparing them with global and ancestral BCoV strains. Additionally, we performed a comparative analysis between wild-type BCoV directly sequenced from the clinical sample (nasal secretion) and the cell culture-adapted strain, utilizing the Sanger method. The field strain and multiple cell passage in cell culture (HRT-18) adapted BCoV strain (BOV19 NS) detected in this study were characterized through molecular and phylogenetic analyses based on partial fragments of 1,448 nt covering the hypervariable region of the S gene. The analyses have demonstrated that different BCoV strains circulating in Brazil, and possibly Brazilian variants, constitute a new genotype (putative G15 genotype). Compared with the ancestral prototype (Mebus strain) of BCoV, 33 nt substitutions were identified of which 15 resulted in non-synonymous mutations (nine transitions and six transversions). Now, compared with the wild-type strain was identified only one nt substitution in nt 2,428 from the seventh passage onwards, which resulted in transversion, neutral-neutral charge, and one substitution of asparagine for tyrosine at aa residue 810 (N810Y).

Spencermartinsiella henanensis fa., sp. nov., a novel yeast species isolated from rotting wood.

Two yeast strains (NYNU 211162 and NYNU 211275) were isolated from rotting wood collected in the Baotianman Nature Reserve, Henan Province, central China. Phylogenetic analysis of the D1/D2 domain of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region revealed that the strains represent a phylogenetically distinct species within the genus Spencermartinsiella. The name Spencermartinsiella henanensis fa., sp. nov. is proposed for this species with holotype CICC 33543T (Mycobank MB 851142). S. henanensis sp. nov. differed by only 3 nt (~0.5 %) substitutions from the closest known species S. europaea NCAIM Y.01817T in the D1/D2 domain, but by 33 nt (~6 %) substitutions, 34 nt (~3.8 %) substitutions, 30 nt (~5.6 %) substitutions and 75 nt (~9.9 %) substitutions in the ITS region and the partial TEF1, COXII and RPB2 genes. Additionally, S. henanensis sp. nov. can be physiologically distinguished from S. europaea by its ability to assimilate inulin, inability to assimilate ethylamine and cadaverine, and incapability of growth at 30 °C.

Foot-and-Mouth Disease Virus Serotype O Exhibits Phenomenal Genetic Lineage Diversity in India during 2018-2022.

In India, widespread foot-and-mouth disease (FMD) outbreaks occurred in 2021. The objective of this study was to identify genetic lineages and evaluate the antigenic relationships of FMD virus (FMDV) isolates gathered from outbreaks reported between 2019 and 2022. Our study shows that the lineages O/ME-SA/Ind2001e and the O/ME-SA/Cluster-2018 were both responsible for the FMD outbreaks on an epidemic scale during 2021. This observation is in contrast to earlier findings that suggested epidemic-scale FMD outbreaks in India are often connected to a single genetic lineage. Additionally, we report here the identification of the O/ME-SA/PanAsia-2/ANT10 sub-lineage in India for the first time, which was connected to two intermittent outbreaks in Jammu and Kashmir. The current study demonstrates that the O/ME-SA/ind2001e lineage has a strong presence outside of the Indian subcontinent. Furthermore, the O/ME-SA/Cluster-2018 was observed to have a wider geographic distribution than previously, and like the O/ME-SA/Ind2001d and O/ME-SA/Ind2001e lineages in the past, it may eventually spread outside of its geographic niche. For O/ME-SA/Ind2001e and O/ME-SA/Cluster-2018, the predicted substitution rate for the VP1 region was 6.737 × 10-3 and 8.257 × 10-3 nt substitutions per site per year, respectively. The time of the most recent common ancestor of the O/ME-SA/Ind2001e and O/ME-SA/Cluster-2018 strains suggests that the viruses possibly emerged during 2003-2011 and 2009-2017, respectively. Recent sightings of the O/ME-SA/PanAsia2/ANT10 virus in India and the O/ME-SA/Ind2001e virus in Pakistan point to possible cross-border transit of the viruses. The results of a two-dimensional viral neutralization test revealed that all of the field isolates were antigenically matched to the currently used Indian vaccine strain O INDR2/1975. These results suggest that the serotype O vaccine strain can protect against outbreaks brought on by all three circulating lineages.

Evolution of avian infectious bronchitis vaccine viruses reisolated from vaccinated broilers

Genetic and phenotypic diversity among infectious bronchitis virus (IBVs) is a key element of the epidemiology of avian infectious bronchitis. Virtually, all studies have focused on the evolution of wild viruses. Live-attenuated vaccine strains deserve a special attention. Two strains of IBV, 2T/17 and 16T/17, detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in samples from H120 vaccinated chickens showing respiratory signs, were genetically analysed targeting the S1 gene. The total RNA was extracted by the classical Trizol method. The whole S1 gene was amplified in an end-point RT-PCR using a specific primer pair, before being sequenced. The obtained sequences were compared to those of vaccine strains and reference viruses and finally subjected to a phylogenetic analysis. The two isolates turned out to be vaccine strains as evidenced by the monophyletic cluster they formed with the H120 vaccine strain (100% bootstrap). Also, they shared with this vaccine strain 99.60% and 99.47% of nucleotides (nt) for strain 2T/17 and 16T/17 respectively. The sequence alignment revealed, for each of the two strains, one non-synonymous nt substitution outside the hypervariable regions: C56T and T39G leading to two amino acid (aa) substitutions: Alv19Val and Cys13Trp, respectively for the strain 2T/17 and 16T/17. The second hypervariable region of strain 2T/17 displayed 2 nt substitutions at positions 345 and 347, resulting in an aa substitution Lys116Thr. As for 16T/17 strain, it harboured two non-synonymous nt substitutions (T353G and T380G), leading to two aa substitutions: Val118Gly and Met127Arg. The chromatogram analysis revealed minor and major peaks at all positions where nt changes were observed. Nucleotides associated with minor peaks matched with those observed in the sequence of the H120 vaccine strain at the same positions, indicating subpopulation selection. Only the substitution T353G results in a mutation as evidenced by the absence of minor peak at this position. Viral subpopulations contained in IBV live-attenuated vaccines are susceptible to be in vivo selected and emerge as persistent pathotype with reversion to virulence. This draws attention to the need of monitoring the evolution of IBV vaccine strains.

Genomic and evolutionary characteristics of G9P[8], the dominant group a rotavirus in China (2016-2018).

G9P[8] became the predominant rotavirus A (RVA) genotype in China in 2012. To evaluate its genetic composition at the whole-genome level, 115 G9P[8] RVA strains isolated from children under 5 years old were sequenced and characterized. All 13 strains in 2016 and 2017 and an additional 54 strains in 2018 were genotyped as G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1. The other 48 strains in 2018 were all genotyped as G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1, with the NSP4 gene characterized as a DS-1-like genotype. The time of the most recent common ancestor (tMRCA) and evolution rates of the VP7, VP4, and NSP4 (E1 and E2) genes of these strains were estimated by Bayesian evolutionary dynamics analysis. We estimated the evolution rates (nt substitutions per site per year) as 1.38 × 10–3 [the 95% highest posterior density (HPD) was 1.09–1.72 × 10–3] for VP7, 0.87 × 10–3 (95% HPD: 0.75–1.00 × 10–3) for VP4, 0.56 × 10–3 (95% HPD: 0.41–0.73 × 10–3) for NSP4-E1, and 1.35 × 10–3 (95% HPD: 0.92–1.86 × 10–3) for NSP4-E2. The tMRCA was estimated to be 1935.4 (95% HPD: 1892.4–1961.3) for VP7, 1894.3 (95% HPD: 1850.5–1937.8) for VP4, 1929.4 (95% HPD: 1892.4–1961.3) for NSP4-E1, and 1969.2 (95% HPD: 1942.2–1985.3) for NSP4-E2. The baseline genetic information in this study is expected to improve our understanding of the genomic and evolutionary characteristics of the rotavirus genome. Furthermore, it will provide a basis for the development of next-generation rotavirus vaccines for humans.

Abstract MP264: Heterogeneous Distribution Of Mutations In Myosin Binding Protein-c Paralogs

Rationale: Myosin binding protein-C ( MYBPC ) is a sarcomeric protein which regulates the force of contraction in striated muscles. Mutations in the MYBPC family of genes, slow skeletal( MYBPC1 ), fast skeletal ( MYBPC2 ) and cardiac ( MYBPC3 ) can result in cardiac and skeletal myopathies, yet their evolutionary pattern, pathogenicity and impact on MYBPC structure remain to be elucidated. Objective: To assess conserved and epigenetic patterns of MYBPC family mutations. Methods and Results: Leveraging a machine learning approach, the Genome Aggregation Database (gnomAD) provided 12353, 6227 and 5279 variants in MYBPC1 , MYBPC2 and MYBPC3 genes, respectively, followed by analysis with the Ensembl variant effector predictor. Missense variants comprised 62-68% of total variants in which the first and second nucleotide (nt) positions in the codons were highly altered, with G/A substitution nt substitution the most predominant. Arginine was the most mutated amino acid, important because most disease-causing mutations in MYBPC are arginine in origin. Domains C5 and C6of MYBPC were found to be hotspots for most mutations in the MYBPC family. A high percentage of truncated mutations in MYBPC3 cause cardiomyopathies. Arginine and glutamate were the top hits in MYBPC1 and MYBPC3, respectively, and tryptophan and tyrosine were the most common among the three paralogs changing to premature stop codons and causing protein truncations at the carboxy terminus. Conclusion: A heterogeneous epigenetic pattern was identified among the three MYBPC paralogs. Genomic databases using machine learning approaches can be used to diagnose and design therapeutics to treat muscle disorders caused by MYBPC mutations.

Natural occurrence of mesta yellow vein mosaic virus and DNA-satellites in ornamental sunflower (Helianthus spp.) in Pakistan.

Weeds and ornamental plants serve as a reservoir for geminiviruses and contribute to their dissemination, genome recombination and/or satellite capture. Ornamental sunflower (Helianthus spp.) plants exhibiting mild leaf curl symptoms were subjected to begomovirus and DNA-satellites isolation. The full-length genome of the isolated begomovirus clone (Od1-A) showed 96.8% nucleotide (nt) sequence identity with mesta yellow vein mosaic virus (MeYVMV; accession no. FR772081) whereas, alphasatellite (Od1-a) and betasatellite (Od1-b) clones showed their highest nt sequence identities at 97.4% and 98.2% with ageratum enation alphasatellite (AEA; accession no. FR772085) and papaya leaf curl betasatellite (PaLCuB; accession. no. LN878112), respectively. The evolutionary relationships, average evolutionary divergence and the recombination events were also inferred. The MeYVMV exhibited 9.5% average evolutionary divergence and its CP and Rep had 9.3% and 12.2%, concomitantly; the alphasatellite and the betasatellite had 8.3% and 5.2%, respectively. The nt substitution rates (site-1 year−1) were found to be 6.983 × 10-04 and 5.702 × 10-05 in the CP and Rep of MeYVMV, respectively. The dN/dS ratio and the Tajima D value of MeYVMV CP demonstrated its possible role in host switching. The absolute quantification of the begomovirus demonstrated that mild symptoms might have a correlation with low virus titer. This is the first identification of MeYVMV and associated DNA-satellites from ornamental sunflower in Pakistan. The role of sequence divergence, recombination and importance of MeYVMV along with DNA-satellites in extending its host range is discussed.

Morphological and molecular data of Hepatozoon ursi in two brown bears (Ursus arctos) in Turkey.

Species of Hepatozoon Miller, 1908 are vector-borne parasites that infect domestic and wild animals worldwide. Hepatozoon ursi Kubo, Uni, Agatsuma, Nagataki, Panciera et al., 2008 was reported from bears (Ursidae) in Japan and India. The present study represents the first report of infection with H. ursi in Turkish brown bears (Ursus arctos Linnaeus) by microscopic and molecular analysis. Two dead brown bears were found in Uzundere and Pasinler districts of Erzurum. Blood and visceral organ (spleen and liver) samples were delivered to laboratory by the Nature Conservation and National Parks officers. Detected gamonts were evaluated based on morphological features and confirmed as gamonts of H. ursi. The size of gamonts and parasitemia were 8.2 × 3.5 μm (6.9-8.7 × 3.0-3.9 μm; n = 12) and 0.6% (6/1000 leukocytes), respectively. The blood and visceral organ samples were positive for species of Hepatozoon by PCR targeting partial sequence of 18S rDNA. Sequence analysis of newly obtained sequences of H. ursi showed 98.8-100% identity with previously sequenced isolates of H. ursi. Sequences of H. ursi from Erzurum were identical to each other and showed 100% identity with isolates of H. ursi from ticks Ixodes ricinus (Linnaeus), Rhipicephalus turanicus Pomerantzev and Hyalomma marginatum Koch collected from two brown bears in Turkey (GenBank accession numbers MN463021, MN463022, MN905023). Analysis of partial sequences of the 18S rRNA gene of H. ursi showed that Turkish isolates differ in NT substitutions found at three different positions [72 (A→G), 537 (A→G) and 570 (A→T)]. This study provides morphological and molecular data of H. ursi infection in brown bears from two districts of Erzurum, Turkey. Further studies are needed to elucidate whether brown bears have any eco-epidemiologic importance in the life cycle of H. ursi in wildlife.

Teunia rosae sp. nov. and Teunia rudbeckiae sp. nov. (Cryptococcaceae, Tremellales), two novel basidiomycetous yeast species isolated from flowers.

Three yeast strains isolated from three flower samples were identified as representing two novel species of Teunia based on molecular phylogenetic analysis and phenotypic comparisons. Strains 12A8 and 21S4 with pink cream colonies and subglobose to globose cells had identical sequences in the ITS and LSU D1/D2 regions, which differed from strain X54 with cream colonies and ovoid to ellipsoidal cells by 6 nt substitutions (1 %) and 9 nt mismatches (1.5 %) in the D1/D2 domains and ITS region, respectively. They could also be distinguished from each other in assimilation of glucitol and salicin, growth at 28 °C and cell fibrillar appendages under scanning electron microscopy. The three strains differed from known species of Teunia by more than 8 nt (1.3 %) and 30 nt (5 %) in the D1/D2 domains and ITS region, respectively. Therefore, the names Teunia rudbeckiae sp. nov. (Holotype CGMCC 2.5840, Mycobank MB 835892) and Teunia rosae sp. nov. (Holotype CGMCC 2.5830, MycoBank MB 835891) are proposed to accommodate strain X54, and strains 12A8 and 21S4, respectively.

Detection and molecular characterization of kobuvirus from diarrheic goats in Minnesota.

Kobuvirus infections are common among humans, rodents, carnivores, pigs, and ruminants. We report herein the complete genome sequence of a novel caprine kobuvirus (MN604700) from diarrheic kids in Minnesota. Whole-genome sequencing revealed a kobuvirus genome of 8,139 nt with a single ORF region encoding a polyprotein of 2,480 amino acids. Further analysis revealed nt substitutions along the genome compared with that of the caprine kobuvirus reference strain, with 93% identity. Phylogenetic analysis indicated that the clade of the caprine kobuvirus was most closely related to porcine kobuviruses rather than bovine or ovine kobuviruses. Using primers designed from this genome, caprine kobuvirus was identified in the stools of other goats. Sanger sequencing of PCR products indicated 3D and VP1 gene nucleotides of this latter strain were 95% and 91% identical with those of MN604700, respectively. There were 35 and 101 nt substitutions in 3D and VP1 genes, respectively. Findings of kobuvirus over a 2-y period may indicate an endemic state, which needs further research. In addition, screening for kobuviruses over large geographic areas is needed to identify the evolutionary connections among different strains.

Naganishia floricola sp. nov., a novel basidiomycetous yeast species isolated from flowers of Sorbaria sorbifolia.

Two yeast strains representing a novel species in the basidiomycetous yeast genus Naganishia were isolated from flowers of Sorbaria sorbifolia collected in Beijing Olympic Forest Park, PR China. Results of multi-gene phylogenetic analysis indicated that the two strains were closely related to the type strains of Naganishia bhutanensis (CBS 6294T) and Naganishia antarctica (CBS 7687T). However, the new isolates differed from N. bhutanensis CBS 6294T by 1.79 % sequence divergence in the D1/D2 domain (11 nt substitutions and three indels), and 2.42 % (15 nt differences and one indel) to N. antarctica CBS 7687T. In the ITS region, the new isolates showed 1.15 % divergence (7 nt substitutions and one indel) to N. bhutanensis CBS 6294T and 0.92 % divergence (5 nt substitutions and no indels) to N. antarctica CBS 7687T. A phylogenetic analysis employing the sequences of six genes (D1/D2 domain of large subunit rDNA, ITS, small subunit rDNA, two subunits of the RNA polymerase II and elongation factor-1α) indicated that the novel species belonged to the genus Naganishia and formed a well-supported clade with N. bhutanensis, N. antarctica and N. indica. Moreover, the two strains differed from their closest relatives by the ability to grow on distinct carbon and nitrogen sources and ability to grow at 30 °C. On the basis of these findings, we propose a novel species in the genus Naganishia (Filobasidiales), Naganishia floricola sp. nov. (holotype CGMCC 2.5856).

Ribavirin induces hepatitis C virus genome mutations in chronic hepatitis patients who failed to respond to prior daclatasvir plus asunaprevir therapy.

Ribavirin (RBV) induces nucleotide (nt) substitutions in hepatitis C virus (HCV) genome nonstructural (NS) regions. Although emergence of drug resistance-associated variants is associated with direct-acting antiviral treatment failure, the effect of RBV on genome substitutions in such patients is unknown. Genotype 1b HCV subgenomic replicon cells were treated with RBV for 120 hours. Six patients with chronic genotype 1b with HCV-infected patients who failed to respond to prior daclatasvir plus asunaprevir (DCV/ASV) therapy were treated with 12 weeks of sofosbuvir and ledipasvir plus RBV after 4 weeks of RBV monotherapy. RBV-induced genome mutations in the HCV NS region (nt3493-9301) in replicon cells and in patients during 4 weeks of RBV monotherapy were analyzed by deep sequencing. RBV-associated G-to-A and C-to-U transitions increased in a dose-dependent manner in HCV replicon cells after the RBV treatment. In patients with prior DCV/ASV treatment failures, the median serum HCV RNA level was 6.25 ± 0.31 log IU/mL at the start of RBV therapy and decreased significantly to 5.95 ± 0.4 log IU/mL (P = .03) after 4 weeks of RBV monotherapy. Although predominant HCV genome substitutions rates were similar between nontreatment and RBV-treatment periods (0.042 and 0.031 per base pair, respectively; P = .248), the frequencies of G-to-A and C-to-U transitions significantly increased after RBV monotherapy. These transitions were enriched, particularly within the HCV NS3 region in all patients. RBV treatment induces G-to-A and C-to-U transitions in the HCV genome even in chronic patients with hepatitis C with prior DCV/ASV treatment failures.

Metschnikowia baotianmanensis f.a., sp. nov., a new yeast species isolated from the gut of the rhinoceros beetle Allomyrina dichotoma.

Four strains, NYNU 15610, NYNU 15612, NYNU 15613 and NYNU 15615, of a novel ascomycetous yeast were isolated from the gut of Allomyrina dichotoma (Coleoptera: Scarabeidae) collected from two different localities in Henan Province, Central PR China. The four strains shared identical sequences in both of the D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer regions. Sequence analyses revealed that this novel species represents a member of the genus Metschnikowia. It differed from its closest known species Metschnikowia zobellii, Metschnikowiaaustralis and Metschnikowia bicuspidata, by 8.4-9.2 % sequence divergence (33-40 nt substitutions and 7-12 gaps over 509 bases) in the D1/D2 sequences. The formation of ascospores was not observed on various sporulation media. In contrast to M. zobellii, M. australis and M. bicuspidata, the novel yeast species was unable to assimilate succinate, ethanol, ethylamine, cadaverine and 10 % NaCl plus 5 % glucose, but was able to grow in vitamin-free medium. The name Metschnikowia baotianmanensis f.a., sp. nov. is proposed to accommodate these strains, with NYNU 15613 as the holotype.

Vishniacozyma changhuana sp. nov., and Vishniacozyma taiwanica sp. nov., two novel yeast species isolated from mangrove forests in Taiwan.

Mangroves grow in the intertidal zone, which alternates between fresh water and sea water, with abundant bioresources. In mangrove habitats, yeasts play an important role in the decomposition of organic matter, and such nutrient cycling has high ecological importance. During a study of the diversity of yeast from Taiwan mangroves, seven strains of basidiomycetous yeasts were isolated and these yeasts represent two novel yeast species belonging to the genus Vishniacozyma. According to the sequences of the D1/D2 domain of large subunit (LSU) rRNA and the internal transcribed spacer (ITS) region, these seven strains could be clearly classified into two groups representing two individual, distinct species. Strains HM5L06, HM6L07, HM11L11 and BJ3S01, differed from their closest relative species Vishniacozyma phoenicis by 4-5 nt substitutions (no gaps) in the sequences of the D1/D2 domain of LSU rRNA and by 23 nt substitutions (10 gaps) in the ITS region. Strains HM6L11, HM7L02 and HM8L19, differed from their closest relative species Vishniacozyma penaeus by 7 nt substitutions (one gap) in the sequences of the D1/D2 domain of LSU rRNA and by 27 nt substitutions (12 gaps) in the ITS region. The scientific names of Vishniacozyma taiwanica sp. nov. and Vishniacozyma changhuana sp. nov. are proposed for these strains. The holotypes are Vishniacozyma taiwanica BCRC 23477T (ex-type HM5L06=CBS 16558; MycoBank number MB837428) and Vishniacozyma changhuana BCRC 23478T (ex-type HM6L11=CBS 16556; MycoBank number MB837429).

Hanseniaspora terricola sp. nov., an ascomycetous yeast isolated from Tibet.

Eight apiculate strains isolated from Tibet, PR China, were identified as Hanseniaspora taiwanica and a novel species of Hanseniaspora based on the sequence analysis of the ITS region, the D1/D2 domains of the LSU rRNA and the translation elongation factor 1-a (TEF1) gene. Among them, four strains with identical sequences of D1/D2 and ITS formed a separate branch from the known Hanseniaspora species in the phylogenetic trees, and differed from the known species by at least 17 (3 %) nucleotide (nt) substitutions in the D1/D2 domains and more than 6 % substitutions and inserts/deletes in the ITS region. The phylogenetic analysis indicated that those four strains represent a novel species of Hanseniaspora, for which the names Hanseniaspora terricola sp. nov. (holotype CGMCC 2.6175T; MycoBank MB 834591) is proposed. The other four strains belonging to H. taiwanica produce spherical, void or fusiform ascospores, which differ from the original description that ascospores are absent.

Mrakia hoshinonis sp. nov., a novel psychrophilic yeast isolated from a retreating glacier on Ellesmere Island in the Canadian High Arctic.

Four strains isolated from sediment sampled at the front of a retreating glacier on northern Ellesmere Island in the Canadian high Arctic, namely JCM 32575T, JCM 32576, JCM 32577 and JCM 32578, belong to a novel psychrophilic basidiomycetous yeast species in the genus Mrakia. Molecular phylogenetic analysis indicated that these strains are most closely related to the type strains of Mrakia aquatica and Mrakianic combsii, but with 8-9 and 7-12 nt substitutions in ITS and in the D1/D2 domain of the LSU rRNA gene, respectively. The strains grew at sub-zero temperatures and in vitamin-free media, with lipase and cellulase highly active even at -3 °C. These characteristics likely allow this yeast species to grow and survive in extremely cold, oligotrophic environments, such as the fronts of retreating glaciers in the high Arctic. The name Mrakia hoshinonis sp. nov. is proposed, with type strain JCM 32575T (UAMH 11969) and MycoBank number MB 825484.

Vishniacozyma ellesmerensis sp. nov., a psychrophilic yeast isolated from a retreating glacier in the Canadian High Arctic.

Two strains of a psychrophilic basidiomycetous yeast species belonging to the genus Vishniacozyma were isolated from sediments and soil at the front of a retreating glacier on northern Ellesmere Island in the Canadian Arctic. Analysis of the large subunit D1/D2 region and the internal transcribed spacer (ITS) regions of the rDNA indicated that these strains represented a novel species. The closest relatives of the novel species were Vishniacozyma globispora and V. dimennae, which exhibited sequence similarities of 82.2 and 81.6 %, respectively, to the ITS region of the novel species, and contained 61 and 57 nt substitutions, respectively, in the D1/D2 domain, in comparison with the novel species. Strains of the novel species were able to grow at sub-zero temperatures and in vitamin-free medium. These characteristics allow the strains to survive and grow in perennially cold, oligotrophic habitats in the Canadian High Arctic. The name Vishniacozyma ellesmerensis sp. nov. is proposed. The type strain is JCM 32573T (=UAMH 11973T=G3-4-8T) and the MycoBank number is MB825501.

The male and female complete mitochondrial genomes of the threatened freshwater pearl mussel Margaritifera margaritifera (Linnaeus, 1758) (Bivalvia: Margaritiferidae)

The complete mitogenomes of one (M-)ale (North America), one Hermaphroditic (Europe), and two (F-)emale (North America and Europe) individuals of the freshwater pearl mussel Margaritifera margaritifera were sequenced. The M-type and F-type (Female and Hermaphroditic) mitogenomes have 17,421 and 16,122 nucleotides, respectively. All with the same content: 13 protein-coding genes, 22 transfer RNA, two ribosomal RNA genes, and one sex-related ORF. The M-type is highly divergent (37.6% uncorrected p-distance) from the F-type mitogenomes. North American and European F-type mitogenomes exhibit low genetic divergence (68 nt substitutions), and the Female and Hermaphroditic European mitogenomes are almost identical, and matching sex-related ORFs.

Mucor chuxiongensis sp. nov., a novel fungal species isolated from rotten wood.

Three strains of a novel mucoralean fungus were isolated from samples of decayed wood, which were collected from three locations near the city of Chuxiong, Yunnan province, China. These isolates were identified as a novel species through comparison of sequences in the ITS sequence, the D1/D2 domains of the LSU rRNA gene, and other taxonomic characteristics. The results demonstrated that these isolates represent a novel mucoralean fungus species belonging to the genus Mucor. The ITS sequence of Mucor chuxiongensissp. nov. differed from its closest relative, Mucor guilliermondii CBS 174.27T, by 13.1 % sequence divergence (39 substitutions and 38 gaps), and the D1/D2 sequences of the novel strains differed by 13 nt substitutions and one gap (1.9 %) from the ex-type strain. On potato dextrose agar, malt extract agar and synthetic mucor agar, the isolates grew slowly below 10 °C, rapidly at 25-30 °C, and could not grow well at 35 °C. The holotype strain of Mucor chuxiongensis sp. nov. is NYNU 174111 (CICC 41666T=CBS 143707T).