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Lamiinae is the most diverse subfamily of longhorned beetles, with about 20,000 described species classified into 80 tribes. Most of the tribes of Lamiinae were proposed during the 19th century and the suprageneric classification of the subfamily has never been assessed under phylogenetic criteria. In this study, we present the first tribal-level phylogeny of Lamiinae, inferred from 130 terminals (representing 46 tribes, prioritizing generic type species of the tribes) and fragments of two mitochondrial and three nuclear markers (cox1, rrnL, Wg, CPS and LSU; 5,024 aligned positions in total). Analyses were performed under Maximum Likelihood and Bayesian methods based on two datasets: a dataset including all taxa available for the study, and a reduced dataset with 111 terminals where taxa only contributing with mitochondrial markers were excluded from the matrix. The monophyly of Lamiinae was corroborated in three of the four analyses and 11 of the 35 tribes with more than one species represented in the analyses were consistently recovered as monophyletic. However, 15 tribes were not retrieved as monophyletic, requiring a revision of their boundaries: Acanthocinini, Acanthoderini, Agapanthiini, Apomecynini, Desmiphorini, Dorcaschematini, Enicodini, Hemilophini, Monochamini, Onciderini, Parmenini, Phytoeciini, Pogonocherini, Pteropliini and Saperdini. Based on these results, when strong support values for paraphyly were recovered, we argue a number of tribe synonymies, including Moneilemini as synonym of Acanthocinini; Onocephalini of Onciderini; Dorcadionini, Gnomini, Monochamini and Rhodopinini of Lamiini; and Obereini and Phytoeciini of Saperdini. Other taxonomic changes proposed in this study based on the criterion of monophyly and supported by morphological characters include the transfer of Tricondyloides and Stenellipsis to Enicodini, and of Dylobolus stat. rest., which is removed as subgenus of Mecas and restituted as genus, to Hemilophini. Furthermore, our analyses suggest that Ostedes and Neohoplonotus should be removed from Acanthocinini and Parmenini, respectively, and Colobotheini should be redefined to encompass several genera currently placed in Acanthocinini., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Myxosporeans (Myxozoa) are eukaryotic parasites, primarily of fish, whose classification is in a state of flux as taxonomists attempt to synthesize the traditional morphology-based system with emerging DNA sequence-based phylogenies. The genus Sphaerospora Thélohan, 1892, which includes pathogenic species that cause significant impacts on fisheries and aquaculture, is one of the most polyphyletic taxa and exemplifies the current challenges facing myxozoan taxonomists. The type species, S. elegans, clusters within the Sphaerospora sensu stricto clade, members of which share similar tissue tropism and long insertions in their variable rRNA gene regions. However, other morphologically similar sphaerosporids lie in different branches of myxozoan phylogenetic trees. Herein, we significantly extend taxonomic sampling of sphaerosporids with SSU+LSU rDNA and EF-2 sequence data for 12 taxa including three representatives of the morphologically similar genus Polysporoplasma Sitjà-Bobadilla et Álvarez-Pellitero, 1995. These taxa were sampled from different vertebrate host groups, biogeographic realms and environments. Our phylogenetic analyses and statistical tests of single and concatenated datasets revealed Sphaerospora s. s. as a strongly supported monophyletic lineage, that clustered sister to the whole myxosporean clade (freshwater+marine lineages). Generally, Sphaerospora s. s. rDNA sequences (up to 3.7 kb) are the longest of all myxozoans and indeed metazoans. The sphaerosporid clade has two lineages, which have specific morphological, biological and sequence traits. Lineage A taxa (marine Sphaerospora spp.) have a single binucleate sporoplasm and shorter AT-rich rDNA inserts. Lineage B taxa (freshwater/brackish Sphaerospora spp.+marine/brackish Polysporoplasma spp.) have 2-12 uninucleate sporoplasms and longer GC-rich rDNA inserts. Lineage B has four subclades that correlate with host group and habitat; all Polysporoplasma species, including the type species, cluster together in one of these subclades. We thus suppress the genus Polysporoplasma and the family Polysporoplasmidae and emend the generic diagnosis of the genus Sphaerospora. The combination of morphological, biological and DNA sequence data applied in this study helped to elucidate an important part of the taxonomic puzzle within the phylum Myxozoa., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Relationships among members of the ciliate subclass Scuticociliatia (Ciliophora, Oligohymenophorea) are largely unresolved. Phylogenetic studies of its orders Pleuronematida and Loxocephalida were initially based on small subunit ribosomal RNA gene (SSU-rDNA) analyses of a limited number of taxa. Here we characterized 37 sequences (SSU-rDNA, ITS-5.8S and LSU-rDNA) from 21 taxonomically controversial members of these orders. Phylogenetic trees constructed to assess the inter- and intra-generic relationships of pleuronematids and loxocephalids reveal the following: (1) the order Loxocephalida and its two families Loxocephalidae and Cinetochilidae are not monophyletic when more taxa are added; (2) the core pleuronematids are divided into two fully supported clades, however, the order Pleuronematida is not monophyletic because Cyclidium glaucoma is closer to Thigmotrichida; (3) the family Pleuronematidae and the genus Schizocalyptra are monophyletic, though rDNA sequences of Pleuronema species are highly variable; (4) Pseudoplatynematum and Sathrophilus are closely related to the subclass Astomatia, while Cinetochilum forms a monophyletic group with the subclass Apostomatia; and (5) Hippocomos falls in the order Pleuronematida and is closely related to Eurystomatellidae and Cyclidium plouneouri. Further, in an effort to provide a better resolution of evolutionary relationships, the secondary structures of ITS2 transcripts and the variable region 4 (V4) of the small subunit ribosomal RNA (SSU-rRNA) are predicted, revealing that ITS2 structures are conserved at the order level while V4 region structures are more variable than ITS2 structures., (Copyright © 2013 Elsevier Inc. All rights reserved.)

We examined the usefulness of the D3 domain and flanking core regions (=D3(+)) of the nuclear large subunit (LSU) ribosomal DNA as a genetic marker for species-level identification and the inference of evolutionary relationships of ixodid ticks. Genetic variation was also examined in relation to the secondary structure of the LSU rDNA. The results revealed a lack of sequence difference in the D3(+) among species of Dermacentor and among some species of Ixodes, demonstrating that this gene region is not suitable as a species marker for all species of ixodid ticks. Of the 45 variable nucleotide positions in the sequence alignment of the D3(+), 23 did not alter the secondary structure of the LSU rDNA, because they occurred in unpaired positions, whereas 16 represented partial or full compensatory changes which maintained the secondary structure. Six deletions in the D3(+) sequence of all Ixodes species examined resulted in a shorter d4_1 helix compared with that of other tick species. The results of the phylogenetic analyses also showed that the D3(+) is of limited value in resolving evolutionary relationships among ixodid ticks. In addition, we also demonstrated that the D3(+) of ascomycete fungi could also be amplified along with, or instead of, the D3(+) of some tick species, depending upon the primers used in PCR. Nonetheless, the D3(+) of the fungal contaminants are readily distinguished from the D3(+) of ixodid ticks because of a shorter length and the absence of helix d4_1 in the secondary structure of the LSU rDNA., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The stringent stress response is vital for bacterial survival under adverse environmental conditions. Obligate intracellular Chlamydia lack key stringent response proteins, but nevertheless can interrupt the cell cycle and enter stasis or persistence upon amino acid starvation. A possible key protein retained is YhbZ, a homologue of the ObgE guanosine triphosphatase (GTPase) superfamily connecting the stringent stress response to ribosome maturation. Curiously, chlamydial YhbZ lacks the ObgE C-terminal domain thought to be essential for binding the large ribosomal subunit. We expressed recombinant Chlamydia abortus YhbZ and showed it to be a functional GTPase, with similar activity to other Obg GTPase family members. As Chlamydia are resistant to genetic manipulation, we performed heterologous expression and gradient centrifugation experiments in Escherichia coli and found that, despite the missing C-terminal domain, C. abortus YhbZ co-fractionates with the E. coli 50S large ribosomal subunit. In addition, overexpression of chlamydial YhbZ in E. coli leads to growth defects and elongation, as reported for other Obg members. YhbZ did not complement an E. coli obgE temperature-sensitive mutant, indicating the C-terminal acidic domain may have an additional role. This data supports a role for YhbZ linking the chlamydial stress response to ribosome function and cellular growth., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

In this study we examine the utility of three mitochondrial (COI, COII, 16S) and two nuclear (CAD and EF-1a) markers for estimating lower-level phylogenetic relationships within the dipteran family Chironomidae. As a test case we use species of the genus Micropsectra and the putatively closely related genera Krenopsectra, Parapsectra and Paratanytarsus. We also examine the phylogenetic evidence for the currently accepted species groups within the genus Micropsectra. In our results, highly variable EF-1a sequences within some species indicate the first find of paralogous gene copies in nematocerous Diptera. Among the other genes, COI is found to have the weakest while CAD contains the strongest phylogenetic signal. The resulting phylogeny displays a well-supported, but paraphyletic Micropsectra with regard to Krenopsectra acuta and five Parapsectra species, indicating taxonomic synonymy of these genera with 100% posterior probability. The genus Parapsectra is polyphyletic within Micropsectra while Paratanytarsus remains monophyletic although with low posterior probability. Micropsectra acuta, M. bumasta, M. fallax, M. nohedensis, M. mendli, M. uliginosa, M. chionophila, M. nana, M. styriaca and M. wagneri will all be new combinations as a consequence of the synonymy., (Copyright © 2010 Elsevier Inc. All rights reserved.)

An insertion of one or two amino acids at the monomer-monomer junctions of polyubiquitin is a distinct and highly conserved molecular character that is shared by two very diverse clades of microeukaryotes, the Cercozoa and the Foraminifera. It has been suggested that an insertion consisting of one amino acid, like that found in foraminiferans and some cercozoans, represents an ancestral state, and an insertion consisting of two amino acids represents a derived state. However, the limited number of cercozoan taxa examined so far limits inferences about the number and frequency of state changes associated with this character over deep evolutionary time. Cercozoa include a very diverse assemblage of mainly uncultivated amoeboflagellates, and their tenuous phylogenetic interrelationships have been based largely on small subunit (SSU) rDNA sequences. Because concatenated datasets consisting of both SSU and large subunit (LSU) rDNA sequences have been shown to more robustly recover the phylogenetic relationships of other major groups of eukaryotes, we employed a similar approach for the Cercozoa. In order to reconstruct the evolutionary history of this group, we amplified twelve LSU rDNAs, three SSU rDNAs, and seven polyubiquitin sequences from several different cercozoans, especially uncultured taxa isolated from marine benthic habitats. The distribution of single amino acid insertions and double amino acid insertions on the phylogenetic trees inferred from the concatenated dataset indicates that the gain and loss of amino acid residues between polyubiquitin monomers occurred several times independently. Nonetheless, all of the cercozoans we examined possessed at least one amino acid insertion between the polyubiquitin monomers, which reinforced the significance of this feature as a molecular signature for identifying members of the Cercozoa and the Foraminifera. Our study also showed that analyses combining both SSU and LSU rDNA sequences leads to improved phylogenetic resolution and statistical support for deeper branches within the Cercozoa., (Copyright © 2010 Elsevier Inc. All rights reserved.)

The new animal phylogeny inferred from ribosomal genes some years ago has prompted a number of radical rearrangements of the traditional, morphology based metazoan tree. The two main bilaterian clades, Deuterostomia and Protostomia, find strong support, but the protostomes consist of two sister groups, Ecdysozoa and Lophotrochozoa, not seen in morphology based trees. Although widely accepted, not all recent molecular phylogenetic analyses have supported the tripartite structure of the new animal phylogeny. Furthermore, even if the small ribosomal subunit (SSU) based phylogeny is correct, there is a frustrating lack of resolution of relationships between the phyla that make up the three clades of this tree. To address this issue, we have assembled a dataset including a large number of aligned sequence positions as well as a broad sampling of metazoan phyla. Our dataset consists of sequence data from ribosomal and mitochondrial genes combined with new data from protein coding genes (5139 amino acid and 3524 nucleotide positions in total) from 37 representative taxa sampled across the Metazoa. Our data show strong support for the basic structure of the new animal phylogeny as well as for the Mandibulata including Myriapoda. We also provide some resolution within the Lophotrochozoa, where we confirm support for a monophyletic clade of Echiura, Sipuncula and Annelida and surprising evidence of a close relationship between Brachiopoda and Nemertea.

The three brown banded land snail genus Humboldtiana (Pulmonata: Humboldtianidae) comprises 50 species of insular distribution from southern Texas to central México; its low vagility and dispersal potential are reflected in high levels of endemism and highly restricted distributions. A current scheme of classification recognizes six subgenera: Polyomphala, Oreades, Gymnopallax, Clydonacme, Aglotrochus, and Humboldtiana (the latter containing three species groups). In the present work, mitochondrial 16S rDNA and partial ITS region and ribosomal large subunit (5.8S+ITS-2+28S) sequences from 26 recognized species and 2 populations of unknown identity were studied to assess phylogenetic relationships of Humboldtiana species using parsimony and Bayesian analysis, and AU test was used to compare the different phylogenetic hypotheses. The same four major clades were recovered by both methods (buffoniana, queretaroana, montezumae and fortis), although none of them support the current scheme of classification, with the only exception being the subgenus Gymnopallax included in the clade buffoniana. The results suggest that speciation patterns observed in the genus Humboldtiana could be explained as a morphostatic radiation.