Ribosomal Unit Research Articles

Mitochondrial genome of Trichagalma acutissimae (Hymenoptera: Cynipoidea: Cynipidae) and phylogenetic analysis

Trichagalma acutissimae (Monzen) (Hymenoptera: Cynipidae) is a major pest of Quercus variabilis Blume in the Taihang Mountains in China. In this study, we sequenced and analyzed the mitochondrial genome (mitogenome) of T. acutissimae. This mitogenome was 16,078 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and 2 ribosomal RNA unit genes (rRNAs). The whole mitogenome exhibited heavy AT nucleotide bias (86.2%). Except for nad4L that started with TTG, all other PCGs started with the standard ATN codon. All 13 PCGs terminate with the stop codon TAA. Phylogenetic analysis showed that T. acutissimae got together with Synergus sp. with high support value, indicating the close relationship of these two genus. All five Cynipoidea species constituted a major clade and formed a sister group to Proctotrupoidea and Chalcidoidea.

Description and phylogenetic analyses of ribosomal transcription units from species of Fasciolidae (Platyhelminthes: Digenea).

Many members of Fasciolidae are common trematodes in cattle, buffaloes, sheep, elephants, pigs, with some capable of infecting humans also. In this study, the complete or near-complete sequences of ribosomal transcription unit (rTU or rDNA), each of Fasciola hepatica (Australia), Fascioloides jacksoni (Sri Lanka), Fasciolopsis buski (Vietnam) and three isolates of F. gigantica (Vietnam), were obtained and characterized. The full length of rDNA for each F. hepatica, 'hybrid' Fasciola sp., Fas. jacksoni and Fa. Buski, was 7657 bp, 7966 bp, 7781 bp and 8361 bp, with the complete intergenic spacer region (IGS) (862 bp, 1170 bp, 987 bp and 561 bp), respectively. The rDNA of two 'pure' F. gigantica isolates from Vietnam was 6794 bp with unsequenced IGS. For 28S rRNA genes the Fasciola spp. are equal, 1958 bp for 18S, 160 bp for 5.8S, 3863 bp and 454 bp for ITS1 but ITS2 differ by one nucleotide (Thymine) (359 or 360 bp). The ITS1 of the sensu lato Fa. buski has some distinguishable features, 286 bp for ITS2, 3862 bp for 28S and four repeat units of 356-361 bp each found in ITS1. The 28S rDNA analysis showed the lowest level of divergence (0-0.57%) between F. hepatica and F. gigantica and higher (2.23-2.62%) and highest (6-6.42%) for Fas. jacksoni and Fasciolopsis, respectively. The tree of 43 strains/species clearly produced a well-supported phylogeny, where 18 fasciolids consistently grouped, forming a discrete Fasciolidae clade, distinct from Philophthalmidae, Echinostomatidae and Echinochasmidae in Echinostomatoidea. Fascioloides jacksoni is outside Fasciola spp.: basal with Fas. magna, as previously demonstrated.

Study of Symbiont Bacteria of <i>Acropora digitifera</i> Coral From Ciletuh Bay, Sukabumi by Using Culture and Molecular Approach

Bacteria are one of the prokaryotic microorganisms that are symbiotic with coral reefs. These microorganisms help corals in secreting mucus layers which are used as a place to live for bacteria and control the presence of pathogenic bacteria in corals. The purpose of this study was to determine the diversity of bacterial communities associated with Acropora digitifera corals in Ciletuh bay, West Java, by culture approach. Sampling was carried out at a depth of 4 meters. Morphology-based identification and molecular approach based on sequence of the small ribosomal unit (16S) rRNA gene were used to determine the bacterial species in the samples. A total of six pure isolates were identified based on morphological observation and the molecular DNA characteristics from three of them were identified with 16S rRNA gene sequences. The identification using 16S rRNA gene showed that the isolate ACD.P4.PH7.P had a close relationship with the BF strain and zb strain of Bacillus flexus (acc number MH569560.1) with a similarity of 85.44 %. While ACD.P4.PH9.P isolate has a close relationship with Bacillus sp. c234 (acc number FJ950647.1) with a similarity of 98.50 %. Additionally, ACD.P4.PH9.K isolates closely related to Bacillus sp. of strain 6RM1 (acc number MK134607.1) with a similarity of 94.78 %. This study also revealed that both microscopic identifications by morphological traits and molecular approach using 16S rRNA gene can be used in bacterial diversity assessment.

The nuclear 18S ribosomal DNAs of avian haemosporidian parasites

BackgroundPlasmodium species feature only four to eight nuclear ribosomal units on different chromosomes, which are assumed to evolve independently according to a birth-and-death model, in which new variants originate by duplication and others are deleted throughout time. Moreover, distinct ribosomal units were shown to be expressed during different developmental stages in the vertebrate and mosquito hosts. Here, the 18S rDNA sequences of 32 species of avian haemosporidian parasites are reported and compared to those of simian and rodent Plasmodium species.MethodsAlmost the entire 18S rDNAs of avian haemosporidians belonging to the genera Plasmodium (7), Haemoproteus (9), and Leucocytozoon (16) were obtained by PCR, molecular cloning, and sequencing ten clones each. Phylogenetic trees were calculated and sequence patterns were analysed and compared to those of simian and rodent malaria species. A section of the mitochondrial CytB was also sequenced.ResultsSequence patterns in most avian Plasmodium species were similar to those in the mammalian parasites with most species featuring two distinct 18S rDNA sequence clusters. Distinct 18S variants were also found in Haemoproteus tartakovskyi and the three Leucocytozoon species, whereas the other species featured sets of similar haplotypes. The 18S rDNA GC-contents of the Leucocytozoon toddi complex and the subgenus Parahaemoproteus were extremely high with 49.3% and 44.9%, respectively. The 18S sequences of several species from all three genera showed chimeric features, thus indicating recombination.ConclusionGene duplication events leading to two diverged main sequence clusters happened independently in at least six out of seven avian Plasmodium species, thus supporting evolution according to a birth-and-death model like proposed for the ribosomal units of simian and rodent Plasmodium species. Patterns were similar in the 18S rDNAs of the Leucocytozoon toddi complex and Haemoproteus tartakovskyi. However, the 18S rDNAs of the other species seem to evolve in concerted fashion like in most eukaryotes, but the presence of chimeric variants indicates that the ribosomal units rather evolve in a semi-concerted manner. The new data may provide a basis for studies testing whether differential expression of distinct 18S rDNA also occurs in avian Plasmodium species and related haemosporidian parasites.

Tracing zoonotic parasite infections throughout human evolution

AbstractParasites are useful pathogens to explore human–animal interactions because they have diverse life cycles that often rely on both as hosts. Moreover, some species are not host specific and are transmitted between animals and humans. Today most emerging infections are zoonoses. Here, we take a specific look at the emergence and re‐emergence of zoonotic parasites throughout hominin evolution and consider evolutionary, cultural, and ecological factors involved in this. We combine genetic studies focused on molecular phylogenetic reconstructions, most often using the ribosomal RNA gene unit and mitochondrial genes from modern parasites, archaeological evidence in the form of preserved parasite eggs and antigens in skeletal and mummified remains, and modern epidemiological data to explore parasite infections throughout hominin evolution. We point out the considerably ancient origins of some key zoonotic parasites and their long coevolutionary history with humans and discuss factors contributing to the presence of many zoonotic parasites in the past and today including dietary preferences, urbanization, waste disposal, and the population density of both humans and domesticated animals.

The complete mitochondrial genome of click beetle Agrypnus sp. (Coleoptera: Elateridae) and phylogenetic analysis

Larvae of click beetles are the most important soil-dwelling agricultural pests and are abundant throughout the world. Agrypnus sp. belongs to the subfamily Agrypninae of Elateridae. Here, we sequenced and annotated the complete mitochondrial genome (mitogenome) of Agrypnus sp., the new representative of the mitogenome of the subfamily. This mitogenome was 16,056 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and two ribosomal RNA unit genes (rRNAs). Gene order was conserved and identical to most other previously sequenced Elateridae. The nucleotide composition biases towards A and T, which together made up 70.8% of the entirety. Phylogenetic analysis based on 13 PCGs sequences showed that Agrypnus sp. got together with the same subfamily species Pyrophorus divergens, Pyrearinus termitilluminans, Ignelater luminosus, and Hapsodrilus ignifer with high support value.

Characterization and phylogenetic analysis of the complete mitochondrial genome of Aphis spiraecola (Hemiptera: Aphididae)

Aphis spiraecola is an important pest of citrus and transmits a number of plant viruses. Here, we sequenced and annotated the complete mitochondrial genome (mitogenome) of A. spiraecola. This mitogenome was 15,465 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and 2 ribosomal RNA unit genes (rRNAs). Gene order was conserved and identical to that of Drosophila yakuba and to most other previously sequenced Aphididae. The whole mitogenome exhibited heavy AT nucleotide bias (81.8%). All 13 PCGs were initiated by the ATN (ATG, ATT and ATA) codon. Except for cox1 and nad4 which end with the incomplete codon T−, all PCGs terminated with the stop codon TAA. Phylogenetic analysis showed that A. spiraecola got together with the same genus species Aphis gossypii and Aphis craccivora with high support value, and Aphis had a close relationship with Schizaphis and Rhopalosiphum.

Mitochondrial genome of Tamarixia radiata (Hymenoptera: Chalcidoidea: Eulophidae) and phylogenetic analysis

Tamarixia radiata plays an important role in biological control of the psyllid Diaphorina citri Kuwayama, vector of the huanglongbing (HLB). In this study, we sequenced and analyzed the mitochondrial genome (mitogenome) of T. radiata, the first mitogenome of species in the family Eulophidae. This mitogenome was 14,752 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs) and two ribosomal RNA unit genes (rRNAs). All 13 PCGs were initiated by the ATN (ATG, ATT, and ATA) codon. Twelve PCGs terminate with the stop codon TAA or TAG except for nad1 which end with the incomplete codon T−. Phylogenetic analysis showed that T. radiata got together with three Pteromalidae species, indicating the close relationship of Eulophidae and Pteromalidae.

Characterisation and phylogenetic analysis of the complete mitochondrial genome of Ctenoplusia albostriata (Lepidoptera: Noctuidae: Plusiinae)

Ctenoplusia albostriata is a pest of composite plants such as Calendula officinalis L. and Dahlia pinnata. In this study, we sequenced and analyzed the complete mitochondrial genome (mitogenome) of C. albostriata. This mitogenome was 15,284 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and 2 ribosomal RNA unit genes (rRNAs). Gene order was conserved and was found to be identical to most other previously sequenced Noctuidae. The whole mitogenome exhibited heavy AT nucleotide bias (80.9%). Except for cox1 started with CGA, all other PCGs started with the standard ATN codons. Most of the PCGs terminated with the stop codon TAA, whereas cox1, cox2 and nad4 end with the incomplete codon T––. Phylogenetic analysis showed that the mitogenome of C. albostriata was similar to C. agnata and C. limbirena, and the subfamily Plusiinae was close to Acronictinae, Heliothinae, Amphipyrinae, Noctuinae, and Hadeninae.

Mitochondrial genome of Diaphorencyrtus aligarhensis (Hymenoptera: Chalcidoidea: Encyrtidae) and phylogenetic analysis

Diaphorencyrtus aligarhensis is an important natural enemy of the psyllid Diaphorina citri Kuwayama, a vector of the huanglongbing (HLB). Here, we sequenced and annotated the mitochondrial genome (mitogenome) of D. aligarhensis. This mitogenome was 16,264 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and two ribosomal RNA unit genes (rRNAs). All 13 PCGs were initiated by the ATN (ATG, ATT, ATA, and ATC) codon. All PCGs terminate with the stop codon TAA except for cox2 and nad1 which end with the incomplete codon T−. Phylogenetic analysis showed that D. aligarhensis got together with the same family species Encyrtus infelix, and Encyrtidae had a close relationship with Agaonidae.

Phylogenetic relationship and characterization of the complete mitochondrial genome of Milionia basalis (Lepidoptera: Geometridae)

Larvae of the geometrid moth Milionia basalis feed exclusively on the podocarp tree, Podocarpus macrophyllus. In this study, we sequenced and analyzed the complete mitochondrial genome (mitogenome) of M. basalis. This mitogenome was 15,901 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and two ribosomal RNA unit genes (rRNAs). The whole mitogenome exhibited heavy AT nucleotide bias (82.2%). Gene order was conserved and identical to most other previously sequenced Geometridae. Most PCGs of M. basalis had the conventional start codons ATN, with the exception of nad1 (TTG) and cox1 (CGA). Except for four genes (cox1, cox2, nad5, and nad4) end with the incomplete stop codon T––, all other PCGs terminated with the stop codon TAA. Phylogenetic analysis showed that M. basalis got together with Apocheima cinerarius, Jankowskia athlete, and four Biston species (B. panterinaria, B. perclara, B. suppressaria, and B. thibetaria).

Complete mitochondrial genome sequence of Tapinoma melanocephalum (Hymenoptera: Formicidae)

Tapinoma melanocephalum is a ubiquitous invasive species and widely distributed in subtropical and tropical regions around the world. Here, we sequenced and annotated the complete mitochondrial genome (mitogenome) of T. melanocephalum. This mitogenome was 15,499 bp long and encoded 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and 2 ribosomal RNA unit genes (rRNAs). Compared to other Formicidae species, gene order of T. melanocephalum was not conserved and one tRNA cluster trnW-trnC-trnY converted to trnW-trnY-trnC. The whole mitogenome exhibited heavy AT nucleotide bias (79.5%). All PCGs started with the standard ATN codons. Except for cox1 and nad5 end with the incomplete codon T−, all PCGs terminated with the stop codon TAA. Phylogenetic analysis showed that T. melanocephalum got together with three same subfamily Dolichoderinae species and one Dorylinae species, indicating the close relationship of Dolichoderinae and Dorylinae.

Insights into the molecular phylogeny of Rhaphidophoridae, an ancient, worldwide lineage of Orthoptera

We investigated the molecular phylogenetic divergence and historical biogeography of cave crickets belonging to the family Rhaphidophoridae (Orthoptera, Ensifera). We used taxa representative of most of the regions embraced by the family, considering samples of Macropathinae from Gondwana land (i.e., Tasmania, Australia, New Zealand, South Africa, and South America); Aemodogryllinae and Rhaphidophorinae from Southern-eastern Asia (i.e., India, Bhutan, China, Philippines and the Sulawesi islands); Dolichopodainae and Troglophilinae from the Mediterranean region and Ceuthophilinae from North America. Based on previous papers, we carried out an analysis of both mitochondrial and nuclear DNA sequences considering the ribosomal RNA units 12S, 16S, 18S, and 28S. To reconstruct phylogeny, we use cladistics, Maximum Likelihood (ML), and Bayesian analyses. All phylogenetic analyses showed the same highly supported topology generally congruent with the classical systematic arrangement at the level of each sub-family but strongly disagree with previous affinity hypotheses between sub-families based on morphological characters. Our results reveal a close affinity between Asiatic and Gondwanian taxa from one hand and between North American and Mediterranean ones from the other hand. Dating estimates indicated that Rhaphidophoridae originated in the Cretaceous period during the Mesozoic era with the ancestral area located both in the northern and southern hemisphere. A possible biogeographic scenario, reconstructed using S-DEC with RASP software, suggested that the current distribution of Rhaphidophoridae might be explained by a combination of both dispersal and vicariance events occurred especially in the ancestral populations. The radiation of Rhaphidophoridae started within the Pangaea, where the ancestor of Rhaphidophoridae occurred throughout an ancestral area including Australia, North America, and the Mediterranean region. The opening of the Atlantic Ocean promoted the divergence of North American and Mediterranean lineages while the differentiation of the southern lineages, spread from Australia, appears to be related to the fragmentation of Gondwana land.

Prevalence and molecular characterisation of Sarcocystis miescheriana and Sarcocystis suihominis in wild boars (Sus scrofa) in Italy.

A sample of the diaphragm was collected from each of 100 wild boars legally hunted in the Val Grande National Park in north-western Italy and examined for the presence of Sarcocystis infection by histological and molecular methods. In histological sections, thick-walled sarcocysts consistent with those of Sarcocystis miescheriana were detected in 32 wild boars. Genomic DNA extracted from diaphragm samples was initially subjected to PCR amplification of the internal transcribed spacer 1 (ITS1) region, and 97 wild boars were found to harbour a Sarcocystis infection at this screening. Selected DNA samples were then subjected to PCR amplification and sequencing of the ITS1 region and the 18S and 28S ribosomal RNA (rRNA) genes of the nuclear ribosomal DNA unit, while all positive samples were subjected to PCR amplification of the mitochondrial cytochrome c oxidase subunit I (cox1) gene. S. miescheriana was identified in 97 wild boars (97%), while the zoonotic Sarcocystis suihominis was identified in one wild boar (1%), which also harboured S. miescheriana. Intra-specific sequence variation was found in all four DNA regions of S. miescheriana examined and in the 18S rRNA gene and ITS1 region of S. suihominis. The partial cox1 gene was amplified and sequenced from 72 isolates of S. miescheriana, yielding 43 haplotypes with pairwise sequence identities of 97.6-99.9%. These haplotypes were 79.1-79.8% identical with the cox1 sequence of S. suihominis. Phylogeny based on cox1 sequences placed S. miescheriana and S. suihominis as sister species within a clade comprising mainly Sarcocystis spp. of ruminants with felids as known or presumed definitive hosts. The same was true for the phylogeny based on 18S rRNA gene sequences.

Basic, simple and extendable kinetic model of protein synthesis.

Protein synthesis is one of the most fundamental biological processes. Despite existence of multiple mathematical models of translation, surprisingly, there is no basic and simple chemical kinetic model of this process, derived directly from the detailed kinetic scheme. One of the reasons for this is that the translation process is characterized by indefinite number of states, because of the structure of the polysome. We bypass this difficulty by applying lumping of multiple states of translated mRNA into few dynamical variables and by introducing a variable describing the pool of translating ribosomes. The simplest model can be solved analytically. The simplest model can be extended, if necessary, to take into account various phenomena such as the limited amount of ribosomal units or regulation of translation by microRNA. The introduced model is more suitable to describe the protein synthesis in eukaryotes but it can be extended to prokaryotes. The model can be used as a building block for more complex models of cellular processes. We demonstrate the utility of the model in two examples. First, we determine the critical parameters of the synthesis of a single protein for the case when the ribosomal units are abundant. Second, we demonstrate intrinsic bi-stability in the dynamics of the ribosomal protein turnover and predict that a minimal number of ribosomes should pre-exists in a living cell to sustain its protein synthesis machinery, even in the absence of proliferation.

Chip-SIP: Stable Isotope Probing Analyzed with rRNA-Targeted Microarrays and NanoSIMS.

Chip-SIP is a stable isotope probing (SIP) method for linking microbial identity and function in mixed communities and is capable of analyzing multiple isotopes (13C, 15N, and 18O) simultaneously. This method uses a high-density microarray to separate taxon-specific 16S (or 18S) rRNA genes and a high sensitivity magnetic sector secondary ion mass spectrometer (SIMS) to determine the relative isotope incorporation of the rRNA at each probe location. Using a maskless array synthesizer (MAS), we synthesize multiple unique sequences to target hundreds of taxa at the ribosomal operational taxonomic unit (OTU) level on an array surface, and then analyze it with a NanoSIMS 50, using its high-spatial resolution imaging capability to generate isotope ratios for individual probes. The Chip-SIP method has been used in diverse systems, including surface marine and estuarine water, rhizosphere, and peat soils, to quantify taxon-specific relative incorporation of different substrates in complex microbial communities. Depending on the hypothesis and experimental design, Chip-SIP allows the user to compare the same community incorporating different substrates, different communities incorporating the same substrate(s), or quantify how a community responds to treatment effects, such as temperature or nutrient concentrations.

Phylogeographic structuring of the amphidromous shrimp Atya scabra (Crustacea, Decapoda, Atyidae) unveiled by range-wide mitochondrial DNA sampling

Species with biological traits favourable to long-distance marine larval dispersal might show a phylogeographic structuring over broad regions, even when they are genetically connected within smaller scales. Here, we evaluated this hypothesis by using the widespread amphidromous shrimp Atya scabra, predicting a genetic discontinuity across biogeographical barriers throughout the Western Gulf of Mexico (WGM), Caribbean Sea (CS), south-western Atlantic (SWA) and eastern Atlantic (EA). Using cytochrome oxidase subunit 1 (COI) and 16S ribosomal unit (16S) gene fragments, we did a phylogeographic assessment and genetic characterisation with Bayesian clustering, AMOVA, haplotype networks and demographic analyses. As predicted, three discrete genetic groups, corresponding to the regions WGM, CS and EA, were uncovered by COI, as well an unpredicted SWA+CS group. The 16S fragment detected a low genetic variation, probably owing to a recent lineage differentiation, which was estimated by the COI molecular clock. We evaluated the role of the biological traits of A. scabra, as well as the consequences of Panama Isthmus closure and Pleistocene glaciation cycles in the lineage isolation of WGM and EA, as well as the genetic connectivity shown within regions and between CS and SWA. Our results highlighted that amphidromous species genetically connected over large scales should be genetically characterised in their wide distribution to provide more comprehensive systematics and to assist decision-making in biological conservation.

Mitochondrial genome of Amphimallon solstitiale (Coleoptera: Scarabaeidae: Melolonthinae) and phylogenetic analysis

To better understand the diversity and phylogeny of Scarabaeidae, we sequenced and annotated the nearly complete mitochondrial genome (mitogenome) of Amphimallon solstitiale (Coleoptera: Scarabaeidae: Melolonthinae). This mitogenome was 13,755 bp long and encoded 13 protein-coding genes (PCGs), 19 transfer RNA genes (tRNAs) and two ribosomal RNA unit genes. The A. solstitiale mitogenome with an A + T content of 69.45% presented a positive AT-skew (0.052) and a negative GC-skew (−0.236). Eleven PCGs started with a typical ATN codon, whereas the remaining two PCGs used CAC (cox1) and GTT (cox2) as the initial codon. All tRNAs had a typical secondary cloverleaf structure, except for trnS1 which lacked the dihydrouridine arm. The Bayesian phylogenetic analysis based on mitogenomic data recovered a phylogeny within Scarabaeidae as (Echirinae + (Scirtifomia + (Aphodiinae + (Melolonthinae + (Rutelinae + Cetoniinae))))).

Sequence clustering in bioinformatics: an empirical study.

Sequence clustering is a basic bioinformatics task that is attracting renewed attention with the development of metagenomics and microbiomics. The latest sequencing techniques have decreased costs and as a result, massive amounts of DNA/RNA sequences are being produced. The challenge is to cluster the sequence data using stable, quick and accurate methods. For microbiome sequencing data, 16S ribosomal RNA operational taxonomic units are typically used. However, there is often a gap between algorithm developers and bioinformatics users. Different software tools can produce diverse results and users can find them difficult to analyze. Understanding the different clustering mechanisms is crucial to understanding the results that they produce. In this review, we selected several popular clustering tools, briefly explained the key computing principles, analyzed their characters and compared them using two independent benchmark datasets. Our aim is to assist bioinformatics users in employing suitable clustering tools effectively to analyze big sequencing data. Related data, codes and software tools were accessible at the link http://lab.malab.cn/∼lg/clustering/.

A molecular phylogeny of the tribe Ochthebiini (Coleoptera, Hydraenidae, Ochthebiinae)

AbstractOchthebiinae, with c. 650 species distributed worldwide, are the second most speciose subfamily of the aquatic beetle family Hydraenidae. They are ecologically the most diverse hydraenid subfamily, with terrestrial species as well as species in almost all types of aquatic habitats, including hypersaline waters. Ochthebiinae include the tribes Ochtheosini (four species in three genera) and Ochthebiini. We provide here the first comprehensive phylogeny of the tribe Ochthebiini, based on 186 species and four subspecies from most genera, subgenera and species groups. We obtained sequence data for a combination of mitochondrial and nuclear gene fragments including the 5′ and 3′ ends of the cytochrome c oxidase subunit 1, the 5′ end of 16S RNA plus the leucine tRNA transfer plus 5′ end of NADH dehydrogenase subunit I, and internal fragments of the large and small ribosomal units. The analyses with maximum likelihood (ML) and Bayesian probabilities consistently recovered a generally well supported phylogeny, with most currently accepted taxa and species groups as monophyletic. We provide a new classification of the tribe based on our phylogenetic results, with six genera: Meropathus Enderlein, Ochthebius Leach, Protochthebius Perkins, Prototympanogaster Perkins, Tympallopatrum Perkins and Tympanogaster Janssens. The genus Ochthebius is here divided into nine subgenera in addition to Ochthebius s.s.: (1) O. (Angiochthebius) Jäch & Ribera; (2) O. (Asiobates) Thomson; (3) O. (Aulacochthebius) Kuwert; (4) O. (Cobalius) Rey; (5) O. (Enicocerus) Stephens; (6) O. (Gymnanthelius) Perkins comb.n.; (7) O. (Gymnochthebius) Orchymont; (8) O. (Hughleechia) Perkins comb.n.; and (9) O. (Micragasma) Sahlberg. Within Ochthebius s.s., 17 species groups are proposed, five of them newly established (3, 9, 11, 13 and 16): (1) andraei; (2) atriceps; (3) corrugatus; (4) foveolatus; (5) kosiensis; (6) lobicollis; (7) marinus; (8) metallescens; (9) nitidipennis; (10) notabilis; (11) peisonis; (12) punctatus; (13) quadricollis; (14) rivalis; (15) strigosus; (16) sumatrensis; and (17) vandykei. We elevated to species rank two subspecies of Ochthebius: O. fallaciosus Ganglbauer stat.n. (former subspecies of O. viridis Peyron) and O. deletus Rey stat.rest. (former subspecies of O. subpictus Wollaston).