Well-supported Monophyletic Clade Research Articles

Multiple, Distinct Intercontinental Lineages but Isolation of Australian Populations in a Cosmopolitan Lichen-Forming Fungal Taxon, Psora decipiens (Psoraceae, Ascomycota).

Multiple drivers shape the spatial distribution of species, including dispersal capacity, niche incumbency, climate variability, orographic barriers, and plate tectonics. However, biogeographic patterns of fungi commonly do not fit conventional expectations based on studies of animals and plants. Fungi, in general, are known to occur across exceedingly broad, intercontinental distributions, including some important components of biological soil crust communities (BSCs). However, molecular data often reveal unexpected biogeographic patterns in lichenized fungal species that are assumed to have cosmopolitan distributions. The lichen-forming fungal species Psora decipiens is found on all continents, except Antarctica and occurs in BSCs across diverse habitats, ranging from hot, arid deserts to alpine habitats. In order to better understand factors that shape population structure in cosmopolitan lichen-forming fungal species, we investigated biogeographic patterns in the cosmopolitan taxon P. decipiens, along with the closely related taxa P. crenata and P. saviczii. We generated a multi-locus sequence dataset based on a worldwide sampling of these taxa in order to reconstruct evolutionary relationships and explore phylogeographic patterns. Both P. crenata and P. decipiens were not recovered as monophyletic; and P. saviczii specimens were recovered as a monophyletic clade closely related to a number of lineages comprised of specimens representing P. decipiens. Striking phylogeographic patterns were observed for P. crenata, with populations from distinct geographic regions belonging to well-separated, monophyletic lineages. South African populations of P. crenata were further divided into well-supported sub-clades. While well-supported phylogenetic substructure was also observed for the nominal taxon P. decipiens, nearly all lineages were comprised of specimens collected from intercontinental populations. However, all Australian specimens representing P. decipiens were recovered within a single well-supported monophyletic clade consisting solely of Australian samples. Our study supports up to 10 candidate species-level lineages in P. decipiens, based on genealogical concordance and coalescent-based species delimitation analyses. Our results support the general pattern of the biogeographic isolation of lichen-forming fungal populations in Australia, even in cases where closely related congeners have documented intercontinental distributions. Our study has important implications for understanding factors influencing diversification and distributions of lichens associated with BSC.

Detection of Latino virus (Arenaviridae: Mammarenavirus) naturally infecting Calomys callidus

Mammarenavirus species are associated with a specific rodent host species, although an increasing number of virus has been associated to more than one host, suggesting that co-evolution is less robust than initially thought. There are few eco-epidemiological studies of South America mammarenaviruses in non-endemic areas of Arenavirus Hemorrhagic Fever, affecting specially our current knowledge about animal reservoirs and virus range and host-virus relations. In Brazil, seven arenavirus species were described in seven different rodent species. Here in we describe a new rodent reservoir species in Brazil related to the previously described Latino mammarenavirus (LATV) MARU strain. Samples of 148 rodents from Mato Grosso state, Brazil were analyzed. Amplification of the glycoprotein precursor gene (GPC) was observed in six Calomys callidus rodents. According to phylogenetic inferences, is observed a well-supported monophyletic clade of LATV from C. callidus and other Clade C mammarenavirus. In addition, the phylogenetic relations of both genes showed a close relation between LATV MARU and Capão Seco strains, two distinct lineages. Additionally, the results obtained in this study point out to a change of scenario and in previously stabilized patterns in the dynamics of South American mammarenaviruses, showing that with more studies in AHF non-endemic or silent areas, more potential hosts for this virus will be discovered.

Phylogeography of Dominican Republic bats and implications for systematic relationships in the Neotropics

The majority (90%) of native terrestrial mammal species living in the Dominican Republic are bats, and two-thirds of these species are endemic to the Caribbean. However, recent molecular studies using DNA barcoding of the mitochondrial cytochrome c oxidase subunit 1 gene have suggested at least a 25% underestimation of biodiversity in bats throughout the world. A recent survey of bats in the Dominican Republic documented 15 of the 18 known species on the island of Hispaniola. Phylogenetic analysis of 132 individuals resulted in well-supported monophyletic species-level clades (maximal bootstrap values) with intraspecific variation ranging from 0% to 4.7% and interspecific variation ranging from 14.1% to 32.5%. A phylogeographic pattern separating the northern and southern Dominican Republic was recovered in 3 species of bats (Macrotus waterhousii, Pteronotus parnellii, and Pteronotus quadridens). The inclusion of broader geographic sampling across the Neotropics indicated that 3 widely distributed species (Eptesicus fuscus, Molossus molossus, and Monophyllus redmani) have high sequence divergence among insular or between insular and continental populations. Further systematic study is needed to identify morphologically cryptic species and their implications for conservation priorities in the Caribbean.

Phylogenetic revision of Camarosporium (Pleosporineae, Dothideomycetes) and allied genera.

A concatenated dataset of LSU, SSU, ITS and tef1 DNA sequence data was analysed to investigate the taxonomic position and phylogenetic relationships of the genus Camarosporium in Pleosporineae (Dothideomycetes). Newly generated sequences from camarosporium-like taxa collected from Europe (Italy) and Russia form a well-supported monophyletic clade within Pleosporineae. A new genus Camarosporidiella and a new family Camarosporidiellaceae are established to accommodate these taxa. Four new species, Neocamarosporium korfii, N. lamiacearum, N. salicorniicola and N. salsolae, constitute a strongly supported clade with several known taxa for which the new family, Neocamarosporiaceae, is introduced. The genus Staurosphaeria based on S. lycii is resurrected and epitypified, and shown to accommodate the recently introduced genus Hazslinszkyomyces in Coniothyriaceae with significant statistical support. Camarosporium quaternatum, the type species of Camarosporium and Camarosporomyces flavigena cluster together in a monophyletic clade with significant statistical support and sister to the Leptosphaeriaceae. To better resolve interfamilial/intergeneric level relationships and improve taxonomic understanding within Pleosporineae, we validate Camarosporiaceae to accommodate Camarosporium and Camarosporomyces. The latter taxa along with other species are described in this study.

A new putative deltapartitivirus recovered from Dianthusamurensis.

Two double stranded RNAs (dsRNA), likely representing the genome of a novel deltapartitivirus, provisionally named carnation cryptic virus 3 (CCV3), were recovered from Dianthus amurensis. The two dsRNAs were 1,573 (dsRNA1) and 1,561 (dsRNA2) bp in size, each containing a single open reading frame (ORF) encoding a 475- and 411-aa protein, respectively. The 475-aa protein contains a conserved RNA dependent RNA polymerase (RdRp) domain which shows significant homology to RdRps of established or putative partitiviruses, particularly those belonging to the genus Deltapartitivirus. However, it shares an amino acid identity of 75% with its closest relative, the RdRp of the deltapartitivirus beet cryptic virus 2 (BCV2), and is <62% identical to the RdRps of other partitiviruses. In a phylogenetic tree constructed with RdRps of selected partitiviruses, CCV3 clustered with BCV2 and formed a well-supported monophyletic clade with known or putative deltapartitiviruses.

Isolation and characterization of microsatellite markers for Sturnira parvidens and cross-species amplification in Sturnira species.

BackgroundSturnira is one of the most species-rich genera in the Neotropics, and it is found from Mexico and the Lesser Antilles to Argentina. This genus forms a well-supported monophyletic clade with at least twenty-one recognized species, as well as several others under taxonomic review. Sturnira parvidens is a widespread frugivorous bat of the deciduous forests of the Neotropics, is highly abundant, and is a major component in fruit dispersal to regenerate ecosystems.MethodsWe used a technique based on Illumina paired-end sequencing of a library highly enriched for microsatellite repeats to develop loci for S. parvidens. We analyzed millions of resulting reads with specialized software to extract those reads that contained di-, tri-, tetra-, penta-, and hexanucleotide microsatellites.ResultsWe selected and tested 14 polymorphic (di, tri, and tetra) microsatellites. All markers were genotyped on 26 different individuals from distinct locations of the distributional area of S. parvidens. We observed medium—high genetic variation across most loci, but only 12 were functionally polymorphic. Levels of expected heterozygosity across all markers were high to medium (mean HE = 0.79, mean HO = 0.72). We examined ascertainment bias in twelve bats of the genus, obtaining null/monomorphic/polymorphic amplifications.DiscussionThe Illumina paired-end sequencing system is capable of identifying massive numbers of microsatellite loci, while expending little time, reducing costs, and providing a large amount of data. The described polymorphic loci for S. parvidens in particular, and for the genus in general, could be suitable for further genetic analysis, including taxonomic inconsistencies, parentage/relatedness analysis, and population genetics assessments.

DNA-Based Identification of Forensically Important Blow Flies (Diptera: Calliphoridae) From India.

Correct species identification is the first and the most important criteria in entomological evidence-based postmortem interval (PMI) estimation. Although morphological keys are available for species identification of adult blow flies, keys for immature stages are either lacking or are incomplete. In this study, cytochrome oxidase subunit 1 (COI) reference data were developed from nine species (belonging to three subfamilies, namely, Calliphorinae, Luciliinae, and Chrysomyinae) of blow flies from India. Seven of the nine species included in this study were found suitable for DNA-based identification using COI gene, because they showed nonoverlapping intra- (0.0-0.3%) and inter-(1.96-18.14%) specific diversity, and formed well-supported monophyletic clade in phylogenetic analysis. The remaining two species (i.e., Chrysomya megacephala (Fabricius) and Chrysomya chani Kurahashi) cannot be distinguished reliably using our database because they had a very low interspecific diversity (0.11%), and Ch. megacephala was paraphyletic with respect to Ch. chani in the phylogenetic analysis. We conclude that the COI gene is a useful marker for DNA-based identification of blow flies from India.

A new putative alphapartitivirus recovered from the powdery mildew fungus Erysiphe palczewskii.

Two double-stranded RNAs (dsRNA) likely representing the genome of a novel alphapartitivirus which we provisionally named Erysiphe palczewskii alphapartitivirus 1 (EpV1) were recovered from the powdery mildew fungus E. palczewskii infecting Sophora japonica in Jingzhou, Hubei province of China. The two dsRNAs, 1955 (dsRNA1) and 1917 (dsRNA2) bp in size, respectively, each contains a single open reading frame (ORF) encoding a 585- and 528-aa protein, respectively. The 585-aa protein contains a conserved RNA-dependent RNA polymerase (RdRp) domain and shows significant homology to RdRps of approved or putative partitiviruses, particularly those belonging to the genus Alphapartitivirus. However, it shares an aa sequence identity lower than 80% with its closest relative, the RdRp of the putative alphapartitivirus Grapevine partitivirus, and lower than 60% with the RdRps of other partitiviruses. In a phylogenetic tree constructed with RdRp aa sequences of selected partitiviruses, the putative virus EpV1 clustered with Grapevine partitivirus and formed a well-supported monophyletic clade with known or putative alphapartitiviruses.

Population dynamics of hepatitis C virus subtypes in injecting drug users on methadone maintenance treatment in China associated with economic and health reform.

The extensive genetic heterogeneity of hepatitis C virus (HCV) requires in-depth understanding of the population dynamics of different viral subtypes for more effective control of epidemic outbreaks. We analysed HCV sequences data from 125 participants in Wuhan, China. These participants were newly infected by subtype 1b (n=13), 3a (n=15), 3b (n=50) and 6a (n=39) while on methadone maintenance treatment (MMT). Bayesian phylogenies and demographic histories were inferred for these subtypes. Participants infected with HCV-1b and 3a were clustered in well-supported monophyletic clades, indicating local subepidemics. Subtypes 3b and 6a strains were intermixed with other Chinese isolates, as well as isolates from other Asian countries, reflecting ongoing across geographic boundary transmissions. Subtypes 1b and 3a declined continuously during the past ten years, consistent with the health and economic reform in China, while subtype 3b showed ongoing exponential growth and 6a was characterized by several epidemic waves, possibly related to the recently growing number of travellers between China and other Asian countries. In conclusion, results of this study suggest that HCV subtype 3b and 6a subepidemics in China are currently not under control, and new epidemic waves may emerge given the rapid increase in international travelling following substantial economic growth.

Esteya Vermicola, a Nematophagous Fungus Attacking the Pine Wood Nematode, Harbors a Bacterial Endosymbiont Affiliated with Gammaproteobacteria.

Symbioses have played pivotal roles in biological, ecological, and evolutionary diversification. Symbiotic bacteria affect the biology of hosts in a number of ways. Esteya vermicola, an endoparasitic nematophagous fungus, has high infectivity in the pine wood nematode (PWN), which causes devastating ecological damage and economic losses in Asia and Europe. An integration of molecular, phylogenetic, and morphological analyses revealed that surface-sterilized E. vermicola with septate hyphae from different geographic locations harbor bacterial endosymbionts. 16S rRNA gene sequences from four fungal strains all clustered in a well-supported monophyletic clade that was the most closely related to Pseudomonas stutzeri and affiliated with Gammaproteobacteria. The existence and intracellular location of endobacteria was revealed by fluorescent in situ hybridization (FISH). Our results showed that endobacteria were coccoid, vertically inherited, as yet uncultured, and essential symbionts. Ultrastructural observations indicated that young and old endobacteria differed in cell size, cell wall thickness, and the degree of reproduction. The results of the present study provide a fundamental understanding of the endobacteria inside E. vermicola and raise questions regarding the impact of endobacteria on the biology, ecology, and evolution of their fungal host.

Identification of Novel Betaherpesviruses in Iberian Bats Reveals Parallel Evolution.

A thorough search for bat herpesviruses was carried out in oropharyngeal samples taken from most of the bat species present in the Iberian Peninsula from the Vespertilionidae, Miniopteridae, Molossidae and Rhinolophidae families, in addition to a colony of captive fruit bats from the Pteropodidae family. By using two degenerate consensus PCR methods targeting two conserved genes, distinct and previously unrecognized bat-hosted herpesviruses were identified for the most of the tested species. All together a total of 42 potentially novel bat herpesviruses were partially characterized. Thirty-two of them were tentatively assigned to the Betaherpesvirinae subfamily while the remaining 10 were allocated into the Gammaherpesvirinae subfamily. Significant diversity was observed among the novel sequences when compared with type herpesvirus species of the ICTV-approved genera. The inferred phylogenetic relationships showed that most of the betaherpesviruses sequences fell into a well-supported unique monophyletic clade and support the recognition of a new betaherpesvirus genus. This clade is subdivided into three major clades, corresponding to the families of bats studied. This supports the hypothesis of a species-specific parallel evolution process between the potentially new betaherpesviruses and their bat hosts. Interestingly, two of the betaherpesviruses’ sequences detected in rhinolophid bats clustered together apart from the rest, closely related to viruses that belong to the Roseolovirus genus. This suggests a putative third roseolo lineage. On the contrary, no phylogenetic structure was detected among several potentially novel bat-hosted gammaherpesviruses found in the study. Remarkably, all of the possible novel bat herpesviruses described in this study are linked to a unique bat species.

Polyphyly of the genus Canoparmelia—uncovering incongruences between phenotype-based classification and molecular phylogeny within lichenized Ascomycota (Parmeliaceae)

Many phenotypical features traditionally used to classify genera in Parmeliaceae and in lichens in general have evolved several times independently, potentially limiting their taxonomic utility. Here, we aim to elucidate evolutionary relationships of Canoparmelia s. lat. among other parmotremoid taxa. A multilocus dataset (ITS, nuLSU and mtSSU rDNA sequences) was gathered and analyzed within a phylogenetic framework. Canoparmelia s. lat. was recovered as highly polyphyletic within the parmelioid clade, and three divergent lineages representing Canoparmelia s. lat. were identified in addition to the previously segregated Crespoa group. Of these, two formed a sister relationship with Parmotrema. However, no apparent diagnostic morphological features were found distinguishing the distinct Canoparmelia s. lat. clades reconstructed in the phylogenetic analyses. As a consequence, we propose to restrict the circumscription of Canoparmelia to clade 1 (i.e. the C. texana group) and to include clades 2 and 3 in Parmotrema. We propose to recognize these well-supported monophyletic clades at subgeneric level. Consequently, the new subgeneric name Parmotrema subgen. Africanae is proposed for clade 3 recovered in this study. Since clade 4, which clusters with the genera Nesolechia and Punctelia, is only represented by a single sequenced specimen, we refrain from proposing any taxonomic changes. The new combinations Parmotrema epileucum, and P. zimbabwense are proposed.

Iningainema pulvinus gen nov., sp nov. (Cyanobacteria, Scytonemataceae) a new nodularin producer from Edgbaston Reserve, north-eastern Australia

A new nodularin producing benthic cyanobacterium Iningainema pulvinus gen nov., sp nov. was isolated from a freshwater ambient spring wetland in tropical, north-eastern Australia and characterised using combined morphological and phylogenetic attributes. It formed conspicuous irregularly spherical to discoid, blue-green to olive-green cyanobacterial colonies across the substratum of shallow pools. Morphologically Iningainema is most similar to Scytonematopsis Kiseleva and Scytonema Agardh ex Bornet & Flahault. All three genera have isopolar filaments enveloped by a firm, often layered and coloured sheath; false branching is typically geminate, less commonly singly. Phylogenetic analyses using partial 16S rRNA sequences of three clones of Iningainema pulvinus strain ES0614 showed that it formed a well-supported monophyletic clade. All three clones were 99.7–99.9% similar, however they shared less than 93.9% nucleotide similarity with other cyanobacterial sequences including putatively related taxa within the Scytonemataceae. Amplification of a fragment of the ndaF gene involved in nodularin biosynthesis from Iningainema pulvinus confirmed that it has this genetic determinant. Consistent with these results, analysis of two extracts from strain ES0614 by HPLC–MS/MS confirmed the presence of nodularin at concentrations of 796 and 1096μgg−1 dry weight. This is the third genus of cyanobacteria shown to produce the cyanotoxin nodularin and the first report of nodularin synthesis from the cyanobacterial family Scytonemataceae. These new findings may have implications for the aquatic biota at Edgbaston Reserve, a spring complex which has been identified as a priority conservation area in the central Australian arid and semiarid zones, based on patterns of endemicity.

Molecular Epidemiology and Spatio-Temporal Dynamics of the H3N8 Equine Influenza Virus in South America

Equine influenza virus (EIV) is considered the most important respiratory pathogen of horses as outbreaks of the disease lead to substantial economic losses. The H3N8 EIV has caused respiratory disease in horses across the world, including South American countries. Nucleotide and deduced amino acid sequences for the complete haemagglutinin gene of the H3N8 EIV detected in South America since 1963 were analyzed. Phylogenetic and Bayesian coalescent analyses were carried out to study the origin, the time of the most recent common ancestors (tMRCA), the demographic and the phylogeographic patterns of the H3N8 EIV. The phylogenetic analysis demonstrated that the H3N8 EIV detected in South America grouped in 5 well-supported monophyletic clades, each associated with strains of different origins. The tMRCA estimated for each group suggested that the virus was circulating in North America at least one year before its effective circulation in the South American population. Phylogenetic and coalescent analyses revealed a polyphyletic behavior of the viruses causing the outbreaks in South America between 1963 and 2012, possibly due to the introduction of at least 4 different EIVs through the international movement of horses. In addition, phylodynamic analysis suggested South America as the starting point of the spread of the H3N8 EIV in 1963 and showed migration links from the United States to South America in the subsequent EIV irruptions. Further, an increase in the relative genetic diversity was observed between 2006 and 2007 and a subsequent decline since 2009, probably due to the co-circulation of different lineages and as a result of the incorporation of the Florida clade 2 strain in vaccines, respectively. The observed data highlight the importance of epidemiological surveillance and the implementation of appropriate quarantine procedures to prevent outbreaks of the disease.

The taxonomic status and sister group relationship of the cardinalfish species Jaydia striatodes (Percomorphaceae: Apogonidae).

Two similar cardinalfish species, Jaydia striatodes and J. striata, were compared morphologically and genetically, using the fragment of cytochrome oxidase subunit I (COI) gene of the mitochondrial DNA. The results confirmed the validity of both species and their sister group relationship. The species formed well-supported monophyletic clades that were distinctly separate with mean sequence divergence of 12.2%. Jaydia striatodes is distinct in having 4-5 + 12-13 gill rakers; 3 + 11-12 developed gill rakers; 9 gill rakers on the first ceratobranchial; 3-9 weak serration at the angle of preopercular edge; and a usually blackish distal half of anal fin. Jaydia striatodes was recorded for the first time from Beibu Gulf, China, and from Vietnam.

Dictyota chalchicueyecanensis sp. nov. (Dictyotales, Phaeophyceae) en el Golfo de México: evidencias moleculares y morfológicas

Background. Species of the Dictyota genus have a high phenotypic plasticity that contrasts with their limited morphological diagnostic characteristics, which has led to some uncertainty in identifying species in the Gulf of Mexico. Recent studies worldwide have focused on Dictyota species phylogenies, which are the basis for contrasting and interpreting the results of this study. Goals. In our research, we found evidence of a new species belonging to the Dictyota genus in three localities in the state of Veracruz, Mexico (Playa Hermosa, Punta Puntillas y Villa Rica). Methods. Phylogenetic and molecular analyses of partial sequences of the psbA and cox1 genes were performed, complemented by morphological analysis. Results. In phylogenetic analysis of partial sequences of psbA and cox1, we established that samples of the new species form a well-supported monophyletic clade. In addition, the genetic divergence values that were obtained are within the interspecific values for the genus, allowing us to segregate the Dictyota chalchicueyecanensis sp. nov. from the rest of the species of this genus. Morphologically this new species is characterized by an erect thallus, light brown and lacking iridescence, stiff to the touch, with a discoid holdfast, rounded apices, dichotomous branching, dentated margins, with minute teeth, triangular to spinose in shape that are directed toward the apices. Sporangia are isolated or grouped on both sides of the thallus and germinate regularly in interdichotomous basal portions. Conclusion. These results allow us to expand the diversity of the genus Dictyota worldwide, incorporating D. chalchicueyecanensis as a probable new species endemic to Mexico.

How many species ofHipposideroshave occurred on Madagascar since the Late Pleistocene?

Populations of the Malagasy Hipposideros commersoni (family Hipposideridae) are threatened by deforestation and hunting. Maximum likelihood and Bayesian analysis of 148 cytochrome b sequences found this species to be paraphyletic and composed of three well-supported monophyletic clades. Clades B and C form a monophyletic lineage that can be referred to H. commersoni; these two clades are separated by 6% sequence variation. Clade A represents a distinct evolutionary lineage separate (9–11% average sequence divergence) from H. commersoni (clades B and C) and is named herein as a new species, Hipposideros cryptovalorona sp. nov. In the phylogeny presented herein, this species is strongly associated with the outgroup taxa Hipposideros gigas and Hipposideros vittatus, both restricted to Africa. External, cranial and dental measurements taken from the same individuals used in the molecular study indicate no clear distinction in morphology amongst these three clades; this includes noseleaf structure and craniodental characteristics. Principal component analyses showed limited separation of the three clades. Comparison to a Quaternary fossil species from north-west Madagascar, Hipposideros besaoka, found little morphological overlap between any of the three clades and this extinct species. Hence, at least three species of Hipposideros have occurred on Madagascar since the Late Pleistocene, two extant (H. commersoni s.s. and H. cryptovalorona sp. nov.) and one extinct (H. besaoka). © 2015 The Linnean Society of London

Phylogenetic relationships of corallinaceae (Corallinales, Rhodophyta): taxonomic implications for reef-building corallines.

A new, more complete, five-marker (SSU, LSU, psbA, COI, 23S) molecular phylogeny of the family Corallinaceae, order Corallinales, shows a paraphyletic grouping of seven well-supported monophyletic clades. The taxonomic implications included the amendment of two subfamilies, Neogoniolithoideae and Metagoniolithoideae, and the rejection of Porolithoideae as an independent subfamily. Metagoniolithoideae contained Harveylithon gen. nov., with H.rupestre comb. nov. as the generitype, and H.canariense stat. nov., H.munitum comb. nov., and H.samoënse comb. nov. Spongites and Pneophyllum belonged to separate clades. The subfamily Neogoniolithoideae included the generitype of Spongites, S.fruticulosus, for which an epitype was designated. Pneophyllum requires reassesment. The generitype of Hydrolithon, H.reinboldii, was a younger heterotypic synonym of H.boergesenii. The evolutionary novelty of the subfamilies Hydrolithoideae, Metagoniolithoideae, and Lithophylloideae was the development of tetra/bisporangial conceptacle roofs by filaments surrounding and interspersed among the sporangial initials.

Molecular evolution of H3N8 equine influenza virus in Argentina

Since 1963, Equine influenza virus (EIV) H3N8 subtype has been responsible for numerous outbreaks of respiratory disease in horses worldwide. The aim of this work was to study the molecular characteristics of H3N8 EIV detected in Argentina, estimate their diversification and infer the potential source of infections. Nucleotide and deduced amino acid (aa) sequences for the complete HA gene of the EIV detected in Argentina between 1985 and 2012 were analyzed along with sequences obtained from the Influenza Research database. Maximum Likelihood (ML) and Bayesian phylogenetic trees were built up. Bayesian coalescent analyses were carried out to estimate the time of the most recent common ancestors (tMRCA) and the dynamic population growth. The phylogenetic trees demonstrate that H3N8 EIV detected in Argentina grouped in 4 well-supported monophyletic clades: Group VIII (G.VIII), South American clade 1 (SA1) and 2 (SA2), and Florida clade 1 (FC1). The viruses circulating in Argentina in 1985 (n1⁄42), together with a Chilean strain, formed G.VIII (tMRCA~1984). The EIVs detected during the period 19931996 (n1⁄45) grouped in the SA1 (tMRCA~1992), composed only by Argentinian strains, and their most probable origin was a Kentucky 1992 strain. The strains circulating in the period 1997-2005 (n1⁄48) grouped in the SA2 (tMRCA~1997), formed a monophyletic group together with a 2006 Chilean strain, and were closely related to California 1997 and Kentucky 1994 strains, feasible ancestors of this group. The viruses detected in 2012 (n1⁄46) grouped within the FC1 (tMRCA~2011) and formed a cluster with the viruses detected in Florida in 2011. The demographic reconstruction showed a remarkable increase in viral diversity in 2006, a slight decrease between 2007 and 2009, and an abrupt decrease since 2009. Compared with A/eq/Fontainebleau/79 (pre-divergent prototype strain), Argentinean strains belonging to G.VIII have 9 aa substitutions: T46I, A93T, R140K, T182S, G222W, V223I, I267V, I187B and L199S, these last two situated at the antigenic site B. In comparison with A/eq/Newmarket/1/93 (American lineage reference strain), the SA 1 Argentinean viruses carried 4 aa substitutions: N132K, Q189N, Q190E and E193K, the last 3 at the antigenic site B, and the SA2 strains showed only one aa substitution at the antigenic site D (I214V). The possible ancestors for SA1 and SA2 also possess the same aa substitutions at the antigenic sites B and D, respectively. EIV Argentinean strains detected in 2012 belonging to FC1 contained 5 aa substitutions in comparison with A/eq/Ohio/01/03 (FC1 reference strain): G7D, R62K, D104N, A138S and V223I. In addition, two different subpopulations can be recognized among them, one with a substitution in K-14A and the other one in K-14T and M70V, this last (M70V) being present only in Argentinean strains. Except for K14A and M70V, the strain considered to be the origin of the group (A/eq/Florida/146609/11) has the same aa substitutions. The obtained results allow us to hypothesize that the outbreaks of equine influenza in Argentina were due to 4 introductions of virus, presumably from North America. The rapid increase in the relative genetic diversity in 2006 could be due to the co-circulation of different lineages, while the abrupt decline observed after 2009 could be related to a subsequent reduction in transmission of EIV H3N8 connected to the incorporation of Florida Clade 2 strains in vaccines.

Tomentella brunneoincrustata, the first described species of the Pisonieae-associated Neotropical Tomentella clade, and phylogenetic analysis of the genus in Mexico

The /tomentella-thelephora lineage is one of the most highly dominant clades among ectomycorrhizal communities worldwide. Despite its importance as a root symbiont, its fruit bodies are inconspicuous and rarely found. Knowledge regarding the diversity of Tomentella in the Neotropics is scarce, and is based largely on environmental samples. Here, we describe a new species, Tomentella brunneoincrustata, including its basidiocarp morphology, mycorrhizal anatomy, and ecology. Because knowledge of Tomentella in Mexico is scarce, we provide the first phylogenetic analysis of this genus in the country. We sequenced the nrITS region of the fungal samples, and sequenced the rbcL and trnL regions to identify the host plant. The phylogenetic analyses were conducted by Bayesian inference. The Bayesian analysis showed that several paraphyletic clades within the lineage /tomentella-thelephora are associated with Pisonieae present across tropical regions of the world. However, the ectomycorrhizae sequences from Puerto Rico, Florida, Dominica, and Mexico constituted a well-supported monophyletic clade that we denote as the “Pisonieae-associated Neotropical Tomentella clade”. Within this clade, T. brunneoincrustata was characterized as follows: a thin crustose, strongly attached to the substrate basidiome; concolorous subiculum, undifferentiated and sterile margin; two types of subiculum hyphae; and small (<8 μm) globose to ellipsoid spores. This species develops in tropical dry forests, where it associates with hosts in the Pisonieae tribe within the Nyctaginaceae. The remaining Tomentella fruit body vouchers collected in temperate forests of Mexico belonged to clades related to T. atramentaria, T. pilosa, T. muricata, T. fuscocinerea, T. stuposa, T. punicea, T. atroarenicolor, T. bryophila, and T. lateritia. Five fruit body vouchers had unique sequences forming independent and unknown clades of Tomentella.