Single Subclade Research Articles

Phytohabitans maris sp. nov., isolated from marine sediment.

A novel actinobacterium, strain ZYX-F-186T, was isolated from marine sediment sampled on Yongxing Island, Hainan Province, PR China. Based on the results of 16S rRNA gene sequence analysis, strain ZYX-F-186T belongs to the genus Phytohabitans, with high similarity to Phytohabitans kaempferiae KK1-3T (98.3 %), Phytohabitans rumicis K11-0047T (98.1 %), Phytohabitans flavus K09-0627T (98.1 %), Phytohabitans houttuyneae K11-0057T (97.9 %), Phytohabitans suffuscus K07-0523T (97.7 %), and Phytohabitans aurantiacus RD004123T (97.7 %). Phylogenetic analysis of 16S rRNA gene sequences showed that the strain formed a single subclade in the genus Phytohabitans. The novel isolate contained meso-diaminopimelic acid, d-glutamic acid, glycine, d-alanine, and l-lysine in the cell wall. The whole-cell sugars were xylose, arabinose, ribose, and rhamnose. The predominant menaquinones were MK-9(H8), MK-9(H6), and MK-9(H4). The characteristic phospholipids were phosphatidylethanolamine, phosphatidylinositol, phosphatidylmethylethanolamine, phosphatidylglycerol, and an unknown phospholipid. The major fatty acids (>5 %) were iso-C16 : 0, anteiso-C17 : 0, and iso-C18 : 0. Genome sequencing showed a DNA G+C content of 71.9 mol%. Low average nucleotide identity, digital DNA-DNA hybridization, and average amino acid identity values demonstrated that strain ZYX-F-186T could be readily distinguished from its closely related species. Based on its phylogenetic, chemotaxonomic, and physiological characteristics, strain ZYX-F-186T represents a novel species of the genus Phytohabitans, for which the name Phytohabitans maris sp. nov. is proposed. The type strain is ZYX-F-186T (=CGMCC 4.8025T=CCTCC AA 2023025T=JCM 36507T).

High-throughput nanopore sequencing of Treponema pallidum tandem repeat genes arp and tp0470 reveals clade-specific patterns and recapitulates global whole genome phylogeny.

Sequencing of most Treponema pallidum genomes excludes repeat regions in tp0470 and the tp0433 gene, encoding the acidic repeat protein (arp). As a first step to understanding the evolution and function of these genes and the proteins they encode, we developed a protocol to nanopore sequence tp0470 and arp genes from 212 clinical samples collected from ten countries on six continents. Both tp0470 and arp repeat structures recapitulate the whole genome phylogeny, with subclade-specific patterns emerging. The number of tp0470 repeats is on average appears to be higher in Nichols-like clade strains than in SS14-like clade strains. Consistent with previous studies, we found that 14-repeat arp sequences predominate across both major clades, but the combination and order of repeat type varies among subclades, with many arp sequence variants limited to a single subclade. Although strains that were closely related by whole genome sequencing frequently had the same arp repeat length, this was not always the case. Structural modeling of TP0470 suggested that the eight residue repeats form an extended α-helix, predicted to be periplasmic. Modeling of the ARP revealed a C-terminal sporulation-related repeat (SPOR) domain, predicted to bind denuded peptidoglycan, with repeat regions possibly incorporated into a highly charged β-sheet. Outside of the repeats, all TP0470 and ARP amino acid sequences were identical. Together, our data, along with functional considerations, suggests that both TP0470 and ARP proteins may be involved in T. pallidum cell envelope remodeling and homeostasis, with their highly plastic repeat regions playing as-yet-undetermined roles.

Phylogenetic comparative analysis: Chemical and biological features of caseins (alpha-S-1, alpha-S-2, beta- and kappa-) in domestic dairy animals.

Caseins determine the physicochemical, physiological, and biological characteristics of milk. Four caseins—alpha-S-1, alpha-S-2, beta, and kappa—were analyzed phylogenetically and in silico and characterized regarding chemical, antimicrobial, and antioxidant features in five dairy animals: Arabian camels, sheep, goats, cattle, and water buffalos. The sequence of full-length amino acids of the four caseins for the five species was retracted from the NCBI GenBank database. Multiple sequence alignment is used to examine further the candidate sequences for phylogenetic analysis using Clustal X and NJ-Plot tools. The results revealed that sheep and goats possess strong similarities (98.06%) because of their common ancestor. The same was observed with cattle and water buffalos (96.25%). The Arabian camel was located in a single subclade due to low similarity in casein residues and compositions with other dairy animals. Protein modeling showed that alpha-S1- and alpha-S2-caseins possess the highest number of phosphoserine residues. The in silico computed chemical properties showed that β-casein recorded highest hydrophobicity index and lowest basic amino acid content, while α-S2-casein showed the opposite. The computed biological parameters revealed that α-S2-casein presented the highest bactericidal stretches. Only Arabian camel β-casein and k-casein showed one bactericidal stretches. The analysis also revealed that β-casein, particularly in Arabian camels, possesses the highest antioxidant activity index. These results support the importance of the bioinformatics resources to determine milk casein micelles' chemical and biological activities.

The Evolution of a Specialized, Highly Virulent Fish Pathogen through Gene Loss and Acquisition of Host-Specific Survival Mechanisms.

ABSTRACTPhotobacterium damselae comprises two subspecies, P. damselae subsp. damselae and P. damselae subsp. piscicida, that contrast remarkably despite their taxonomic relationship. The former is opportunistic and free-living but can cause disease in compromised individuals from a broad diversity of taxa, while the latter is a highly specialized, primary fish pathogen. Here, we employ new closed curated genome assemblies from Australia to estimate the global phylogenetic structure of the species P. damselae. We identify genes responsible for the shift from an opportunist to a host-adapted fish pathogen, potentially via an arthropod vector as fish-to-fish transmission was not achieved in repeated cohabitation challenges despite high virulence for Seriola lalandi. Acquisition of ShdA adhesin and of thiol peroxidase may have allowed the environmental, generalist ancestor to colonize zooplankton and to occasionally enter in fish host sentinel cells. As dependence on the host has increased, P. damselae has lost nonessential genes, such as those related to nitrite and sulfite reduction, urea degradation, a type 6 secretion system (T6SS) and several toxin-antitoxin (TA) systems. Similar to the evolution of Yersinia pestis, the loss of urease may be the crucial event that allowed the pathogen to stably colonize zooplankton vectors. Acquisition of host-specific genes, such as those required to form a sialic acid capsule, was likely necessary for the emergent P. damselae subsp. piscicida to become a highly specialized, facultative intracellular fish pathogen. Processes that have shaped P. damselae subsp. piscicida from subsp. damselae are similar to those underlying evolution of Yersinia pestis from Y. pseudotuberculosis.IMPORTANCE Photobacterium damselae subsp. damselae is a ubiquitous marine bacterium and opportunistic pathogen of compromised hosts of diverse taxa. In contrast, its sister subspecies P. damselae subsp. piscicida (Pdp) is highly virulent in fish. Pdp has evolved from a single subclade of Pdd through gene loss and acquisition. We show that fish-to-fish transmission does not occur in repeated infection models in the primary host, Seriola lalandi, and present genomic evidence for vector-borne transmission, potentially via zooplankton. The broad genomic changes from generalist Pdd to specialist Pdp parallel those of the environmental opportunist Yersinia pseudotuberculosis to vector-borne plague bacterium Y. pestis and demonstrate that evolutionary processes in bacterial pathogens are universal between the terrestrial and marine biosphere.

Genetic variability of Begonia longifolia Blume from Indonesia based on Nuclear DNA internal transcribed spacer (ITS) sequence data

Abstract.Ningrum WDA, Atmaja MB, Daryono BS, Purnomo. 2020. Genetic variability of Begonia longifolia Blume from Indonesia based on Nuclear DNA internal transcribed spacer (ITS) sequence data. Biodiversitas 21: 5778-5785. Begonia longifolia Blume is a member of the Begoniaceae family which has the widest distribution in Asia including Indonesia. Diversity studies on B.longifolia from Indonesia were mainly based on morphological characters. The genetic diversity analysis of this pantropical species in Indonesia has never been reported yet. This research aimed to study genetic variability of B. longifolia in Indonesia based on nuclear DNA internal transcribed spacer (ITS) sequence data. Twelve living specimens were from Sumatra (Lampung), Java (Sukabumi, Purwokerto), Bali (Buleleng, Gianyar, Jembrana, Tabanan), and Lombok. The 831-839 length sequence was aligned using Mesquite software. Reconstruction of the phylogenetic tree was done by MEGA7 software using Neighbor-Joining method with Kimura-2-Parameter. The 827 fixed sites showed a genetic distance ranged from 0 to 1.6%. Among the 827 sites compared, 802 sites were invariable, 19 sites were variable (11 singleton variable sites and 8 parsimony informative sites) and 6 sites were gaps. The phylogenetic tree showed 1 clade and 6 sub-clades. Specimens from Sumatra and Java formed a single sub-clade, separate with five sub-clades from Bali. Specimen from Lombok belong to one of the sub-clades of the Bali specimens. All specimens can be divided into 2 groups: Sumatra-Java and Bali-Lombok group.

Systematics of a survivor: the Cambrian kingstoniid trilobite Blountia Walcott,1916 across the Marjuman-Steptoean (Guzhangian-Paibian) extinction intervalin Laurentian North America.

The Cambrian (Marjuman-Steptoean; Guzhangian-Paibian) kingstoniid trilobite Blountia Walcott, 1916 is distributed widely in shelf strata of Laurentian North America. Species known from Marjuman formations were lost at the mass extinction at the end of that stage. New species entered the succession during and after the extinction interval, only to disappear within the Aphelaspis Zone of the lower part of the Steptoean Stage. Steptoean species and several uppermost Marjuman (Crepicephalus Zone) species are treated in this monograph. New collections and revision of type and other archival material increase the number of species in Steptoean strata from two to six. Phylogenetic analysis supports monophyly of Blountia and Maryvillia Walcott, 1916; Blountina Lochman, in Lochman Duncan, 1944 is retained as a monotypic taxon. Steptoean species do not form a single subclade within the cladogram, so there is no evidence for a simple monophyletic radiation following the end-Marjuman extinction. New species are Blountia angelae, B. morgancreekensis, B. nevadensis, B. newfoundlandensis, and B. tennesseensis.

The nervous and circulatory systems of a Cretaceous crinoid: preservation, palaeobiology and evolutionary significance

AbstractFeatherstars, comatulid crinoids that shed their stalk during their ontogeny, are the most species‐rich lineage of modern crinoids and the only ones present in shallow water today. Although they are of considerable palaeontological interest as a ‘success story’ of the Mesozoic Marine Revolution, their fossil record is relatively species‐poor and fragmentary. New Spanish fossils of the Cretaceous featherstar Decameros ricordeanus preserve the shape and configuration of nervous and circulatory anatomy in the form of infilled cavities, which we reconstruct from CT scans. The circulatory system of D. ricordeanus was relatively extensive and complex, implying a pattern of coelomic fluid flow that is unique among crinoids, and the peripheral parts of the nervous system include linkages both to the circulatory system and to the surface of the body. A phylogenetic analysis (the first to include both living and fossil featherstars and which includes characters from internal anatomy) recovers D. ricordeanus among the lineage of featherstars that includes Himerometroidea, Tropiometra and ‘Antedonoidea’, among others. D. ricordeanus is larger than almost any modern featherstar, and its elaborate coelomic morphology appears to be a consequence of positive allometry. All featherstars with coelomic diverticula are shown to belong to a single comatulid subclade, and this feature may constitute a synapomorphy of that group. Some preservation of cavities corresponding to soft tissue is probably not exceptional in fossil crinoids, providing an opportunity to study the diversity and evolution of extinct anatomical systems typically only preserved in Lagerstätten.

Klebsiella quasipneumoniae Provides a Window into Carbapenemase Gene Transfer, Plasmid Rearrangements, and Patient Interactions with the Hospital Environment.

Several emerging pathogens have arisen as a result of selection pressures exerted by modern health care. Klebsiella quasipneumoniae was recently defined as a new species, yet its prevalence, niche, and propensity to acquire antimicrobial resistance genes are not fully described. We have been tracking inter- and intraspecies transmission of the Klebsiella pneumoniae carbapenemase (KPC) gene, blaKPC, between bacteria isolated from a single institution. We applied a combination of Illumina and PacBio whole-genome sequencing to identify and compare K. quasipneumoniae from patients and the hospital environment over 10- and 5-year periods, respectively. There were 32 blaKPC-positive K. quasipneumoniae isolates, all of which were identified as K. pneumoniae in the clinical microbiology laboratory, from 8 patients and 11 sink drains, with evidence for seven separate blaKPC plasmid acquisitions. Analysis of a single subclade of K. quasipneumoniae subsp. quasipneumoniae (n = 23 isolates) from three patients and six rooms demonstrated seeding of a sink by a patient, subsequent persistence of the strain in the hospital environment, and then possible transmission to another patient. Longitudinal analysis of this strain demonstrated the acquisition of two unique blaKPC plasmids and then subsequent within-strain genetic rearrangement through transposition and homologous recombination. Our analysis highlights the apparent molecular propensity of K. quasipneumoniae to persist in the environment as well as acquire carbapenemase plasmids from other species and enabled an assessment of the genetic rearrangements which may facilitate horizontal transmission of carbapenemases.

Monitoring of the field application of Metarhizium anisopliae in Brazil revealed high molecular diversity of Metarhizium spp in insects, soil and sugarcane roots

The use of Metarhizium against sugarcane spittlebugs in Brazil is one of the most successful and long lasting biological control programs using entomopathogenic fungus in the world. However, studies to monitor the fate of this fungus on the sugarcane agroecosystem are rare, especially with respect to its persistence, efficacy in pest control and impact on the local populations of Metarhizium. The present study aimed at documenting the efficacy and persistence of M. anisopliae strain ESALQ1604 in a sugarcane field by using microsatellite molecular markers. The species diversity of Metarhizium was characterized in insects, soil and sugarcane roots in a sprayed and an unsprayed plot. Although the infection rates were not very high (≤ 50%), the applied strain was recovered from spittlebugs after 7, 30 and 60 days’ post-application, but accounted for only 50%, 50% and 70.5% of all insects killed by M. anisopliae, respectively. All haplotypes from spittlebug were associated with a single subclade of M. anisopliae. The highest haplotype diversity was found in soil (h = 0.989) and in the smallest in spittlebug (h = 0.779). Metarhizium robertsii, M. anisopliae, M. brunneum; one taxonomically unassigned lineage was found in soil and only M. brunneum and M. anisopliae were isolated from roots. This study revealed the great diversity of Metarhizium spp. in the sugarcane agroecosystem and the importance of the local population of M. anisopliae on spittlebugs management.

Phylogenetic relationships among the Rangeomorpha: the importance of outgroup selection and implications for their diversification

The Rangeomorpha are the oldest, most diverse, and most disparate clade of Ediacaran macrofossils. Easily identifiable by their self-similar branching pattern, they occupied epibenthic niche space ranging from the lowest-tiered and recumbent taxa up to metre-long upright fronds. A phylogenetic analysis using the largest and most complete character set known for this group scored for 14 separate taxa was undertaken to resolve their internal relationships and test previous hypotheses of their evolutionary and ecological history. Owing to the lack of consensus on the relationship amongst Ediacaran clades, several permutations with different potential outgroup taxa were performed. Across these analyses, there is a strong signal for an upright frondose ancestral state for this clade, likely displaying primary branches that were double-sided, nonrotated, with the lower-tiered and recumbent forms being derived members of a single subclade. This has implications on the life history reconstruction as well as taxonomic implications for this clade and the origins of large multicellular life in the late Ediacaran.

The invasion biology of the invasive earwig, Forficula auricularia in Australasian ecosystems

The European earwig, Forficula auricularia, is a cosmopolitan insect endemic to Europe, West Asia and North Africa, which has invaded many temperate regions of the world including Australia and New Zealand. F. auricularia has been shown to be a complex of morphologically identical, reproductively isolated lineages that possess two distinct clades of mitochondrial DNA. Entomological collection data, historical literature and further field collections were used to develop a greater understanding of Australian F. auricularia’s invasion biology and its current distribution. Genetic analysis of F. auricularia collected from Australia and New Zealand using two mitochondrial genes (COI and a fragment overlapping parts of the COI -COII genes) was also undertaken. To identify the possible source populations of the Australasian invasion these sequences were compared to those from 16 locations within Britain and continental Europe. All Australasian populations were shown to be of the clade B lineage. Tasmanian and New Zealand populations consist of a single subclade comprised of only 4 and 1 haplotypes respectively. The Australian mainland populations also contained a second subclade consisting of up to 11 haplotypes indicating that multiple introductions possibly occurred on the Australian mainland. Comparison of mitochondrial genomes from Australasian and European populations showed the Australian mainland subclade to be most closely related to Portuguese haplotypes, and the Tasmanian and New Zealand clade closely related to those in Brittany, France. No European haplotypes perfectly matched the Australasian sequences. Therefore, the original source populations are still to be identified with harbours on the Iberian Peninsula’s western coast and those on the English Channel likely candidate areas.

Phenotypic Innovation and Adaptive Constraints in the Evolutionary Radiation of Palaeozoic Crinoids

To better understand the patterns and processes shaping large-scale phenotypic diversification, I integrate palaeobiological and phylogenetic perspectives to investigate a ~200-million-year radiation using a global sample of Palaeozoic crinoid echinoderms. Results indicate the early history of crinoid diversification is characterized by early burst dynamics with decelerating morphologic rates. However, in contrast with expectation for a single “early burst” model, morphospace continued to expand following a slowdown in rates. In addition, I find evidence for an isolated peak in morphologic rates occurring late in the clade’s history. This episode of elevated rates is not associated with increased disparity, morphologic novelty, or the radiation of a single subclade. Instead, this episode of elevated rates involved multiple subclade radiations driven by environmental change toward a pre-existing adaptive optimum. The decoupling of morphologic disparity with rates of change suggests phenotypic rates are primarily shaped by ecologic factors rather than the origination of morphologic novelty alone. These results suggest phenotypic diversification is far more complex than models commonly assumed in comparative biology. Furthermore, palaeontological disparity patterns are not a reliable proxy for rates after an initial diversifying phase. These issues highlight the need for continued synthesis between fossil and phylogenetic approaches to macroevolution.

Phylogenetic structure and ancestry of Korean clawed salamander, Onychodactylus koreanus (Caudata: Hynobiidae)

Onychodactylus koreanus, a hynobiid salamander species endemic to the Korean Peninsula, can be regarded as a strict ecological specialist, probably vulnerable to anthropogenic environmental modifications and climate change. We used mitochondrial cytochrome b gene to analyze the genetic diversity and phylogenetic structure of O. koreanus from 19 populations collected in an attempt to cover its major distribution within South Korea. A total of 76 haplotypes of O. koreanus obtained in our analyses could be subdivided into three phylogenetic clades, KR, NE and SE. Clade KR haplotypes occur in most of the regions throughout the Korean Peninsula with four distinct subclades (KR I–IV). Clade NE and SE haplotypes were only observed in two populations YY and YS, respectively. Haplotype sharing was scarce even among populations in geographical proximity, and most of the populations were represented by a single clade or subclade, indicating the low level of gene flow among populations. O. koreanus likely originated from the historical southward dispersal of its ancestral lineages following divergence from Chinese O. zhaoermii that was recovered as the sister of O. koreanus in our phylogenetic analysis. Our results have critical implications for the taxonomic status of O. koreanus and its long-term management plan.

Evolutionary Origin of Euglena.

Euglenids (Excavata, Discoba, Euglenozoa, Euglenida) is a group of free-living, single-celled flagellates living in the aquatic environments. The uniting and unique morphological feature of euglenids is the presence of a cell covering called the pellicle. The morphology and organization of the pellicle correlate well with the mode of nutrition and cell movement. Euglenids exhibit diverse modes of nutrition, including phagotrophy and photosynthesis. Photosynthetic species (Euglenophyceae) constitute a single subclade within euglenids. Their plastids embedded by three membranes arose as the result of a secondary endosymbiosis between phagotrophic eukaryovorous euglenid and the Pyramimonas-related green alga. Within photosynthetic euglenids three evolutionary lineages can be distinguished. The most basal lineage is formed by one mixotrophic species, Rapaza viridis. Other photosynthetic euglenids are split into two groups: predominantly marine Eutreptiales and freshwater Euglenales. Euglenales are divided into two families: Phacaceae, comprising three monophyletic genera (Discoplastis, Lepocinclis, Phacus) and Euglenaceae with seven monophyletic genera (Euglenaformis, Euglenaria, Colacium, Cryptoglena, Strombomonas, Trachelomonas, Monomorphina) and polyphyletic genus Euglena. For 150 years researchers have been studying Euglena based solely on morphological features what resulted in hundreds of descriptions of new taxa and many artificial intra-generic classification systems. In spite of the progress towards defining Euglena, it still remains polyphyletic and morphologically almost undistinguishable from members of the recently described genus Euglenaria; members of both genera have cells undergoing metaboly (dynamic changes in cell shape), large chloroplasts with pyrenoids and monomorphic paramylon grains. Model organisms Euglena gracilis Klebs, the species of choice for addressing fundamental questions in eukaryotic biochemistry, cell and molecular biology, is a representative of the genus Euglena.

Disentangling Phylogenetic Relationships in a Hotspot of Diversity: The Butterworts (Pinguicula L., Lentibulariaceae) Endemic to Italy.

The genus Pinguicula (Lentibulariaceae) consists of about 100 carnivorous species, also known as butterworts. Eleven taxa are endemic to Italy, which represents a biodiversity hotspot for butterworts in Europe. The aim of our study was to provide a phylogenetic framework for the Italian endemics, in order to: a) investigate the relationships between species in this group; b) evaluate their actual taxonomic value. To achieve this, we analysed all the taxa endemic to Italy, along with several other species, by means of ITS nrDNA analysis. Our results clarify the relationships between Italian endemics and other Pinguicula taxa identifying a basal polytomy defined by five clades. All of the Italian endemics (with the exception of P. lavalvae) fall within a single large clade, which includes P. vulgaris and allied species. Among them, P. poldinii represents the most isolated lineage. Other taxa show strong molecular similarities and form a single subclade, although their taxonomic ranks can be retained. Pinguicula lattanziae sp. nov., seemingly endemic to Liguria (NW Italy), is also described.

Low symbiont diversity as a potential adaptive strategy in a marginal non-reefal environment: a case study of corals in Hong Kong

Symbiosis with genetically diverse Symbiodinium has been shown to affect host coral physiological responses to environmental stresses. Hong Kong, located in a subtropical region, is a marginal environment for coral growth largely due to its wide annual temperature fluctuation with low mean winter sea water temperature (~16 °C) and variable salinity conditions. The symbiont diversity in Hong Kong corals is therefore worth investigating to enrich our understanding on symbioses in marginal and fluctuating environments. Examination of 56 scleractinian coral species and five soft coral species using denaturing gel gradient electrophoresis of the internal transcribed spacer region 2 found only five distinct subclades of Symbiodinium with C1 the dominant type occurring in all but one scleractinian coral and all soft coral species investigated. C15 and C21 Symbiodinium were found in Porites spp. and Montipora peltiformis, respectively, both of which are vertical transmitters. D8-12 was found in Oulastrea crispata, a stress-tolerant species, and D1 in a single sample of Goniastrea aspera. No spatial differences in Symbiodinium composition were found among different regions of Hong Kong. Seasonal monitoring of tagged Platygyra acuta and Porites spp. colonies also revealed no changes in their symbiont types despite wide ranges of in situ temperature fluctuation. Hong Kong scleractinian corals hosted a remarkably low symbiont diversity compared with corals in the surrounding regions. The predominance of a single subclade, C1 Symbiodinium, suggests that this subclade is best acclimatized to local fluctuating conditions and/or low winter temperature. Forming symbiosis with the best acclimatized symbiont, instead of with a diverse group of symbionts with different physiological performances, either sequentially or simultaneously, may be a strategy used by Hong Kong corals to cope with stressful conditions.

Genetic changes found in a distinct clade of Enterovirus D68 associated with paralysis during the 2014 outbreak.

Enterovirus D68 (EV-D68) caused a severe respiratory illness outbreak in the United States in 2014. Reports of acute flaccid myelitis (AFM)/paralysis (AFP) in several independent epidemiological clusters of children with detectable EV-D68 have raised concerns that genetic changes in EV-D68 could be causing increased disease severity and neurological symptoms. To explore the potential link between EV-D68 genetic variations and symptom changes, we performed a series of comparative genomic analyses of EV-D68 2014 outbreak isolate sequences using data and analytical tools in the Virus Pathogen Resource (ViPR; www.viprbrc.org). Our results suggest that (1) three distinct lineages of EV-D68 were co-circulating in 2013 and 2014; (2) isolates associated with AFM/AFP belong to a single phylogenetic subclade – B1; (3) the majority of isolates from the B1 subclade have 21 unique substitutions that distinguish them from other isolates, including amino acid substitutions in the VP1, VP2, and VP3 capsid proteins and the 3D RNA-dependent RNA polymerase, and nucleotide substitutions in the internal ribosome entry sequence (IRES); (4) at 12 of these positions, B1 isolates carry the same residues observed at equivalent positions in paralysis-causing enteroviruses, including poliovirus, EV-D70 and EV-A71. Based on these results, we hypothesize that unique B1 substitutions may be responsible for the apparent increased incidence of neuropathology associated with the 2014 outbreak.

Molecular epidemiological analyses of Cryptosporidium parvum virus 1 (CSpV1), a symbiotic virus of Cryptosporidium parvum, in Japan

We show that Cryptosporidium parvum virus 1 (CSpV1), a member of the family Partitiviridae, genus Cryspovirus that can infect Cryptosporidium parvum, is a new candidate for high-resolution tool for tracing C. parvum. CSpV1 was detected in all C. parvum-positive samples tested. Phylogenetic analysis of dsRNA1 sequence from CSpV1 can distinguish infected areas of C. parvum on the national level. Sequences detected in samples from Iwate prefecture and other islands (Tanegashima, and Okinawa) belonged to a single clade. This system can differentiate the samples from Hokkaido and south part of Japan as well as from other countries. Samples from Iwate, Tanegashima, and Okinawa belonged to a single subclade, respectively. Therefore, the CSpV1 dsRNA sequences reflect the regional distribution of their host and have potential as a high-resolution tool to trace C. parvum IIaA15G2R1 subtype.

Phylogenetic diversity of Brazilian Metarhizium associated with sugarcane agriculture

Biological control of spittlebugs with Metarhizium (Hypocreales: Clavicipitaceae) in Brazilian sugarcane is an example of effective pest management. However, little is known about the richness, distribution and ecology of Metarhizium species in Brazilian agroecosystems. We investigated Metarhizium diversity within a collection of 96 Brazilian isolates from spittlebugs and other insects, strains used for spittlebug control and soil isolates from sugarcane and other field crops and pristine habitats. A multilocus phylogenetic analysis of 5′-TEF and nuclear intergenic loci MzFG543igs and MzIGS3 yielded robust support for current species limits of the two most abundant taxa, Metarhizium anisopliae and Metarhizium robertsii, and the resolution of two lineages that lie beyond currently recognized species limits in this complex. With a single exception, all isolates from insects belong to a single subclade of M. anisopliae. These data will serve as resources about Metarhizium biodiversity for insect biological control initiatives in South America.

Re-identifying Grateloupia yangjiangensis (Rhodophyta, Halymeniaceae) based on morphological observations, life history and rbcL sequence analyses

On the basis of morphological observations, life history and molecular phylogeny, Grateloupia yangjiangensis, which is similar to G. filicina, G. orientalis, G. catenata, and G. ramosissima in appearance, was re-examined. The results are as follows: (1) the auxiliary-cell ampullae of G. yangjiangensis were of Grateloupia type, thalli was fleshy and gelatinous in texture, and the erect axes were compressed; the cortex was 0.25–0.30 mm thick, consisting of five to seven outer layers, and there were five inner layers of triangular or stellate cells; (2) there was no filamentous stage in the development of the carpospores; (3) the ribulose-1,5-bisphosphate carboxylase/oxygenase gene (rbcL) sequence of four G. yangjiangensis examined showed that there was no intergeneric divergence among them, and for the phylogenetic tree, four sequences of G. yangjiangensis formed a single monophyletic subclade within the large Grateloupia clade of Halymeniaceae. In conclusion, G. yangjiangensis was a single species within the genus Grateloupia. This research provided criterion for identification and cultivation of G. yangjiangensis.