High Nodal Support Research Articles

The complete mitochondrial genome of Brachytarsina amboinensis (Diptera: Hippoboscoidea: Streblidae) provides new insights into phylogenetic relationships of Hippoboscoidea.

The family Streblidae is a significant grouping of dipteran insects within the superfamily Hippoboscoidea, which parasitizes the body surface of bats. With the global spread of bat-related pathogens in recent years, Streblidae has gained increasing attention due to its potential for pathogen transmission. A sample of Brachytarsina amboinensis was sequenced on the B. amboinensis were obtained, compared with available Streblidae mitogenomes, and the phylogeny of Hippoboscoidea was reconstructed. The results indicate that the mitochondrial genome of B. amboinensis exhibits a relatively high degree of conservation, with an identical gene count, arrangement, and orientation as the ancestral insect's genome. Base composition analysis revealed a strong bias towards A and T in the base composition. Selection pressure analysis indicated strong purifying selection acting on cox1. Pairwise genetic distance analysis showed that cox1 evolved at a relatively slow rate. Regarding phylogenetic relationships, the constructed phylogenetic trees using Bayesian inference and Maximum Likelihood methods supported the monophyly of the Hippoboscoidea, Glossinidae, Hippoboscidae, and Nycteribiidae clades, with high nodal support values. Our research confirmed the paraphyly of the families Streblidae. In the familial relations between Nycteribiidae and Streblidae, New World Streblidae share a closer kinship with Nycteribiidae. This contrasts with prior findings which indicated that Old World Streblidae share a closer kinship with Nycteribiidae. This study not only enhances the molecular database for bat flies but also provides a valuable reference for the identification and phylogenetic analysis of Streblidae.

Assessment of nine markers for phylogeny, species and haplotype identification of Kappaphycus species and Eucheuma denticulatum (Solieriaceae, Rhodophyta)

ABSTRACT Molecular studies have contributed to the taxonomy of carrageenan-producing Kappaphycus spp. and Eucheuma denticulatum. However, unresolved species complexes and the lack of standardization in the use of genetic markers impede the identification of specimens and the delineation of a robust taxonomic framework. Here, nine molecular markers (cox1, cox2–3 spacer, cox2, cox3, COB, ITS, psbA, UPA and rbcL) were used to generate a multilocus phylogeny for 113 fresh eucheumatoid samples and four herbarium specimens. Analyses of species delineation and genetic distances confirmed the monophyly of currently accepted taxa. These analyses suggest that clades previously reported as K. striatus KS1 and KS2 are conspecific, and that E. denticulatum EDA ‘spinosum’ and EDB ‘endong/cacing’ are also conspecific. The results also unveiled possible new taxa from Hawaii and Indonesia. Each molecular marker and combinations thereof were assessed with regard to species identification, ease of amplification and sequencing, and haplotype characterization. All genetic markers recorded at least 94% success in the amplification and sequencing of fresh specimens, with cox1 being the most phylogenetically informative. Automatic partitioning, phylogenetic and tree-based assessments showed cox1, cox2–3 spacer, cox2 and rbcL were able to correctly identify species while cox1+ rbcL, COB+rbcL, cox2+ rbcL or cox1+ COB+rbcL trees best represented the phylogeny with consistently high nodal support. Among individual markers, cox1 identified the greatest number of haplotypes, while UPA, partial rbcL (750 bp), ITS, cox3 and cox2–3 spacer were able to retrieve information from herbarium specimens of 12–16 years of age. These molecular results provide a basis for a database essential for the taxonomic framework, cultivar development and germplasm conservation of eucheumatoids.

Morphological observation, molecular identification and evolutionary analysis of Hydatigera kamiyai found in Neodon fuscus from the Qinghai-Tibetan plateau

Hydatigera kamiyai (H. kamiyai) is a new species within Hydatigera that has recently been resurrected. Voles and cats are hosts of H. kamiyai and have a certain impact on its health and economy. Moreover, the Qinghai-Tibetan plateau (QTP) is a research hotspot representing Earth's biodiversity, as its unique geographical environment and climatic conditions support the growth of a variety of mammals and provide favorable conditions for various parasites to complete their life history. The aim of this study was to reveal the phylogenetic relationships and divergence times of H. kamiyai strains isolated from Neodon fuscus on the QTP using morphological and molecular methods. In this study, we morphologically observed H. kamiyai and sequenced the whole mitochondrial genome. Then, we constructed phylogenetic trees with the maximum likelihood (ML) and Bayesian inference (BI) methods. The GTR alternative model was selected for divergence time analysis. These data demonstrated that the results were consistent with the general morphological characteristics of Hydatigera. The whole genome of H. kamiyai was 13,822 bp in size, and the A + T content (73%) was greater than the G + C content (27%). The Ka/Ks values were all <1, indicating that all 13 protein-coding genes (13 PCGs) underwent purifying selection during the process of evolution. The phylogenetic tree generated based on the 13 PCGs, cytochrom oxidase subunit I (COI), 18S rRNA and 28S rRNA revealed close phylogenetic relationships between H. kamiyai and Hydatigera, with high node support for the relationship. The divergence time based on 13 PCGs indicated that H. kamiyai diverged approximately 11.3 million years ago (Mya) in the Miocene. Interestingly, it diverged later than the period of rapid uplift in the QTP. We also speculated that H. kamiyai differentiation was caused by host differentiation due to the favorable living conditions brought about by the uplift of the QTP. As there have been relatively few investigations on the mitochondrial genome of H. kamiyai, our study could provide factual support for further studies of H. kamiyai on the QTP. We also emphasized the importance of further studies of its hosts, Neodon fuscus and cats, which will be important for further understanding the life cycle of H. kamiyai.

Integrative Taxonomy of Prosogonotrema bilabiatum Vigueras, 1940 (Digenea: Sclerodistomidae): A Parasite in Atlantic Spadefish Chaetodipterus faber (Broussonet, 1782) (Acanthuriformes: Ephippidae) from Brazil.

The present work aims to expand the knowledge of the digenean species Prosogonotrema bilabiatum (Sclerodistomidae), a parasite of Chaetodipterus faber (Acanthuriformes) from Brazil, with an integrative taxonomic approach, using light microscopy, scanning electron microscopy, histology, and molecular biology. Forty-one digenean specimens were stained with hydrochloric carmine for morphological studies. Eleven parasites were dehydrated through a graded ethanol series, critical point dried with carbon dioxide, and coated with gold for scanning electron microscopy analysis. Four specimens were processed following histological routine and stained with hematoxylin and eosin and Gomori trichrome. DNA extracted was amplified using 28S partial primer D1-D3. Maximum likelihood and Bayesian inference were performed for phylogenetic analysis. Morphometric and morphological data of the specimens studied ranged in accordance as observed in previous descriptions of the species. Observations from scanning electron microscopy and histology corroborated with those observed in stained whole mounts. Molecular analysis showed that specimens of P. bilabiatum from Brazil clustered with another two sequences of this species fromdifferent hosts and localities, with a high node support value. The integrative taxonomic approach allowed to record and describe new characteristics of P. bilabiatum related to the tegument, the structure and the arrangement of its tissues. The use of molecular markers confirmed that specimens identified as P. bilabiatum from different hosts and localities are all conspecific. Further studies, mainly molecular with less conserved genetic markers, should be carried out to better understand the phylogenetic relationships of Prosogonotrema with Hemiuroidea.

First report of the complete mitochondrial genome of 3 beetles (Coleoptera: Scarabaeidae) harming Gastrodia elata (Asparagales: Orchidaceae).

Gastrodia elata Blume, a valuable traditional Chinese medicine with significant clinical and nutritional importance, is a fungal heterotrophic orchid. We present the first report of the mitochondrial genome structure and characteristics of 3 Scarabaeidae pests affecting G. elata: Sophrops peronosporus Gu & Zhang, Anomala rufiventris Kollar & Redtenbacher, and Callistethus plagiicollis Fairmaire. Each mitogenome contained 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), 2 ribosomal RNAs (rRNAs), and a control region, with no gene rearrangements observed. All 21 tRNAs, except trnS1 that lacks a dihydrouridine, had a stable cloverleaf secondary structure. Maximum likelihood and Bayesian inference analyses based on the 13 PCGs produced 2 topologically similar phylogenetic trees, both of with high nodal support. Larvae of these Scarabaeidae pests cause substantial damage by gnawing on the tubers and roots of G. elata, leading to reduced yield and compromised quality. These findings contribute to phylogenetic studies of Scarabaeidae, expand knowledge of G. elata pests, and offer valuable reference materials for their identification and control.

Phylogenomic analysis of Tachinidae (Diptera: Calyptratae: Oestroidea): a transcriptomic approach to understanding the subfamily relationships.

Tachinidae is the second most species-rich family of Diptera. It comprises four subfamilies, and all of its members have parasitoid habits. We present the first phylogenomic analysis of Tachinidae using transcriptomic data, based on 30 species. We constructed four datasets: three using translated data at the amino acid level (100% coverage, with 106 single-copy protein-coding genes; 75% coverage, with 1359 genes; and 50% coverage, with 1942 genes). The trees were estimated by analysing four matrices using maximum likelihood and maximum parsimony inferences, and only minor differences were found among them. Overall, our topologies are well resolved, with high node support. Polleniidae is corroborated as a sister group to Tachinidae. Within Tachinidae, our results confirm the hypothesis (Phasiinae + Dexiinae) + (Tachininae + Exoristinae). Phasiinae, Dexiinae and Exoristinae are recovered as monophyletic, and Tachininae as polyphyletic. Once again, the tribe Myiophasiini (Tachininae) composes a fifth lineage, clade sister to all the remaining Tachinidae. The Neotropical tribe Iceliini, formerly in Tachininae, is recovered within Exoristinae, sister to Winthemiini. In general, our results are congruent with recent phylogenetic studies that include tachinids, with the important confirmation of the subfamilial relationships and the existence of a fifth lineage of Tachinidae.

Characterization of the complete mitochondrial genome of the longhorn beetle, Batocerahorsfieldi (Coleoptera, Cerambycidae) and its phylogenetic analysis with suitable longhorn beetles.

Mitochondrial genome analysis is an important tool for studying insect phylogenetics. The longhorn beetle, Batocerahorsfieldi, is a significant pest in timber, economic and protection forests. This study determined the mitochondrial genome of B.horsfieldi and compared it with the mitochondrial genomes of other Cerambycidae with the aim of exploring the phylogenetic status of the pest and the evolutionary relationships among some Cerambycidae subgroups. The complete mitochondrial genome of B.horsfieldi was sequenced by the Illumina HiSeq platform. The mitochondrial genome was aligned and compared with the existing mitochondrial genomes of Batoceralineolata and B.rubus in GenBank (MF521888, MW629558, OM161963, respectively). The secondary structure of transfer RNA (tRNA) was predicted using tRNAScan-SE server v.1.21 and MITOS WebSever. Thirteen protein-coding genes (PCGs) and two ribosomal RNA gene sequences of 21 longhorn beetles, including B.horsfieldi, plus two outgroups, Dryopsernesti (Dryopidae) and Heterocerusparallelus (Heteroceridae), were analyzed. The phylogenetic tree was constructed using maximum likelihood and Bayesian inference methods. In this study, we successfully obtained the complete mitochondrial genome of B.horsfieldi for the first time, which is 15 425 bp in length. It contains 37 genes and an A + T-rich region, arranged in the same order as the recognized ancestor of longhorn beetles. The genome of B.horsfieldi is composed of 33.12% A bases, 41.64% T bases, 12.08% C bases, and 13.16% G bases. The structure, nucleotide composition, and codon usage of the new mitochondrial genome are not significantly different from other longhorn mitochondrial genomes. Phylogenetic analyses revealed that Cerambycidae formed a highly supported single clade, and Vesperidae was either clustered with Cerambycidae or formed a separate clade. Interestingly, B.horsfieldi, B.rubus and B.lineolata were clustered with Monochamus and Anoplophora species in both analyses, with high node support. Additionally, the VesperidaeSpiniphilusspinicornis and Vesperussanzi and the 19 Cerambycidae species formed a sister clade in the Bayesian analysis. Our results have produced new complete mitogenomic data, which will provide information for future phylogenetic and taxonomic research, and provide a foundation for future relevant research.

An 1896 specimen helps clarify the phylogenetic placement of the Mexican endemic Hooper’s deer mouse

Hooper’s deer mouse, Peromyscus hooperi, is the sole member of the Peromyscus hooperi species group. This species is endemic to México where it is restricted to the grassland transition zone in the states of Coahuila, Zacatecas, and San Luis Potosí. Previous studies using mitochondrial and nuclear genes (Cytb, Adh1-I2, Fgb-I7 and Rbp3) did not resolve the phylogenetic relationships of this relatively poorly known species. It was hypothesized that P. hooperi is sister to P. crinitus, and these two taxa are related to P. melanotis, P. polionotus, P. maniculatus, P. keeni, P. leucopus, P. gossypinus, P. eremicus, P. californicus, and Osgoodomys banderanus. Based on morphological characters, karyotypes, and allozymes, P. hooperi does not align with either subgenera Haplomylomys or Peromyscus. However, its unique characteristics (e. g., phallus, karyotype) have been recognized, and therefore it has been retained as its own species group. To better resolve the phylogenetic placement of P. hooperi, we performed target-enrichment and high-throughput sequencing and obtained several thousand nuclear ultraconserved elements and a complete mitogenome from a specimen collected in 1896 by Nelson and Goldman in Coahuila, México. We compared these data with 21 other species of neotomines using genome-wide data. Contrary to previous studies, we found high nodal support for the placement of P. hooperi as sister to a clade that includes Podomys floridanus, Neotomodon alstoni, Habromys simulatus, H. ixtlani, Peromyscus mexicanus, P. megalops, P. melanophrys, P. perfulvus, P. aztecus, P. attwateri, P. pectoralis, and P. boylii. We dated a Pliocene divergence of P. hooperi from its sister group at approximately 3.98 mya, and after the split of P. crinitus at ca. 4.31 mya from other peromyscines. We demonstrated that genome-wide data improve the phylogenetic signal, independently of taxon sampling, for a phylogenetically problematic species such as P. hooperi. We recommend that future genomic studies expand taxon sampling, including members of the subgenus Haplomylomys, to confirm the phylogenetic relationships of P. hooperi and the genetic status of its populations.

Tetrathyridia in an endemic lizard from Chile: molecular evidence for South America.

Mesocestoides is a controversial tapeworm with significant lack of data related to systematics and life cycles. This helminth has an indirect life cycle with vertebrates, mostly carnivorous mammals, as definitive hosts. Theoretically, a coprophagous arthropod would be the first intermediate host, and herptiles, mammals, and birds, which prey on these insects, would represent the second intermediate hosts. However, recent evidence suggests that this life cycle would require only two hosts, with no arthropods involved. In the Neotropics, although there are records of mammals and reptiles as hosts for Mescocestoides, no molecular analyses have been performed. This work aimed to record an additional intermediate host and molecularly characterize the isolated larvae. Thus, 18 braided tree iguanas (Liolaemus platei) from Northern Chile were collected and dissected during 2019. One lizard was parasitized by three morphotypes of larvae compatible with tetrathyridia of Mescocestoides. To achieve its specific identity, a molecular approach was performed: 18S rRNA and 12S rRNA loci were amplified through cPCR. The inferred phylogenies confirmed the morphological diagnosis and stated that all morphotypes were conspecifics. The sequences for both loci formed a monophyletic clade with high nodal support, representing a sister taxon to Mescocestoides clade C. This study represents the first molecular characterization of any taxon of Mescocestoides from the Neotropics. Future surveys from potential definitive hosts would help to elucidate its life cycle. Furthermore, an integrative taxonomic approach is required in additional studies from the Neotropical region, which would contribute to a better understanding of the evolutionary relationships of this genus.

Complete mitochondrial genomes from museum specimens clarify millipede evolution in the Eastern Arc Mountains

AbstractThe Eastern Arc Mountains in Tanzania represent a hotspot for biological diversity of global importance. The level of endemism is high, and Eastern Arc biodiversity has been studied extensively in vertebrates and invertebrates, including millipedes. However, millipede evolution is vastly understudied at the molecular level. Therefore, we used next-generation ‘shotgun’ sequencing to obtain mitochondrial genome sequences of 26 museum specimens, representing six genera and 12 millipede species found across the Eastern Arc Mountains. Bayesian and maximum likelihood methods yielded consistent topologies with high node support, confirming a high level of congruence between molecular and morphological analyses. The only exception was a Tropostreptus sigmatospinus individual from Zanzibar, which was placed outside an otherwise monophyletic group consisting of mainland individuals of the same assumed species. For two species with a distribution across several mountain blocks (Tropostreptus sigmatospinus and Tropostreptus hamatus), each mountain population represents a distinct monophyletic lineage. In contrast, we also observe that distinct species exist sympatrically in the same montane forests, indicative of older speciation events that are not defined by current forest distribution. Our results are important for understanding speciation mechanisms in montane rain forests and highlight that ethanol-preserved invertebrates exhibit a tremendous potential for genomic analyses.

Sphaeromyxa cornuti n. sp., a New Species of Myxosporean Infecting the Gallbladder of the Moorish Idol, Zanclus cornutus (Linnaeus, 1758) from Lakshadweep Waters.

The present study describes a new species of myxosporean, Sphaeromyxa cornuti n. sp. infecting the gallbladder of the Moorish idol, Zanclus cornutus (Linnaeus 1758) collected from Lakshadweep waters of the Arabian Sea. Fish were collected using traps and cages. The morphology of mature spores recovered from the gallbladder of Z. cornutus was studied under Nomarski Differential Interference Contrast (DIC) optics. The molecular and phylogenetic analyses were based on SSU rDNA. Sphaeromyxa cornuti n. sp. is characterized by arcuate myxospores with tapering extremities and round ends in valvular, and slightly sigmoid in sutural views (19.2-24.7 µm × 4.1-5.7µm). The two polar capsules are unequally elongate-ovoid in shape and positioned at opposite ends of the spore (6.2-9.7µm × 1.7-2.6µm). Each encloses an irregularly folded, ribbon-like polar tubule, which is oriented parallel to polar capsule axis. In molecular and phylogenetic analyses, the present myxosporean revealed significant differences with related forms and clustered together with S. hellandi within the 'incurvata' group of the Sphaeromyxa clade with high nodal support. Morphological, morphometric, molecular and phylogenetic differences between our material and previously described species of Sphaeromyxa, along with host and geographic variations indicate that the present myxosporean is unique and the name Sphaeromyxa cornuti n. sp. is proposed. This forms the first report of a myxosporean parasite-infecting Z. cornutus.

Phylogenetic Implication of Large Intergenic Spacers: Insights from a Mitogenomic Comparison of Prosopocoilus Stag Beetles (Coleoptera: Lucanidae).

Simple SummaryInsect mitochondrial genomes (mitogenomes) show high diversity in some lineages. In the mitogenome of some Coleoptera species, a large intergenic spacer (IGS) has been identified. However, very little is known about mitogenomes of lucanid beetles. In this work, to provide further insight into the phylogenic relationships among species in lucanid beetles (genus Prosopocoilus), two Prosopocoilus species (Prosopocoilus castaneus and Prosopocoilus laterotarsus) were newly sequenced and comparatively analyzed. Significantly, the two newly sequenced Prosopocoilus species contained a large IGS located between trnI and trnQ. Our phylogenomic analyses showed that P. castaneus and P. laterotarsus were clustered in a clade with typical Prosopocoilus species (Prosopocoilus confucius, Prosopocoilus blanchardi, and Prosopocoilus astacoides). These results provide valuable data for the future study of the phylogenetic relationships in this genus.To explore the characteristics of mitogenomes and discuss the phylogenetic relationships within the genus Prosopocoilus, the mitogenomes of two species (P. castaneus and P. laterotarsus) were newly sequenced and comparatively analyzed. The arrangement of the mitogenome in these two lucanid beetles was the same as that in the inferred ancestral insect, and the nucleotide composition was highly biased towards A + T as in other lucanids. The evolutionary rates of 13 protein-coding genes (PCGs) suggested that their evolution was based on purifying selection. Notably, we found evidence of the presence of a large IGS between trnI and trnQ genes, whose length varied from 375 bp (in P. castaneus) to 158 bp (in P. laterotarsus). Within the large IGS region, a short sequence (TAAAA) was found to be unique among these two species, providing insights into phylogenomic reconstruction. Phylogenetic analyses were performed using the maximum likelihood (IQ-TREE) and Bayesian (PhyloBayes) methods based on 13 protein-coding genes (PCGs) in nucleotides and amino acids (AA) from published mitogenomes (n = 29). The genus Prosopocoilus was found to constitute a distinct clade with high nodal support. Overall, our findings suggested that analysis of the characteristics of the large IGS (presence or absence, size, and location) in mitogenomes of the genus Prosopocoilus may be informative for the phylogenetic and taxonomic analyses and for evaluation of the genus Prosopocoilus, despite the dense sampling materials needed.

Complete mitochondrial genome of the earthworm Amynthas seungpanensis (Clitellata: Megascolecidae)

Amynthas seungpanensis Song and Paik, 1970 (Clitellata: Megascolecidae) is an endemic and ecologically important species in Korea; however, knowledge regarding its genetic characteristics is lacking. In this study, we sequenced the complete mitochondrial genome (mitogenome) of A. seungpanensis. The mitogenome is 15,085-bp long and contains 13 protein-coding genes (PCGs), 2 rRNA genes, 22 tRNA genes, and 1 major non-coding region, the control region. This mitogenome has an arrangement identical to that observed in the mitogenomes of most earthworms, and all 37 genes are transcribed from the same directional strand. All 13 PCGs have the same start codon (ATG). Six PCGs (COI, COII, ND6, CytB, ATP6, and ND4) end with TAA and ND4 ends with TAG, whereas the remaining genes exhibit an incomplete stop codon, T. The A/T content of the complete mitogenome is 65.8%; however, it varies among regions and genes as follows: control region, 77.8%; srRNA genes, 65.8%; lrRNA genes, 68.8%; tRNA genes, 64.2%; and PCGs, 65.0%. Results of phylogenetic analyses suggested the family Megascolecidae to be a monophyletic group with the highest nodal support, whereas the genus Amynthas, which belongs to Megascolecidae, was determined to be non-monophyletic.

Comparison of mitogenomes of three Petalocephala species (Hemiptera: Cicadellidae: Ledrinae) and their phylogenetic analysis.

Ledrinae is a unique group of leafhoppers with a distinct appearance. Petalocephala is the largest Ledrinae genus that is difficult to identify except by dissecting the male genitals. To date, research on Ledrinae is relatively less compared with other leafhoppers. Therefore, to better understand this group, we sequenced and analyzed three complete Petalocephala mitochondrial genomes. We comparatively analyzed these general Petalocephala genomic features (including size, AT content, AT/GC skew, 13 protein-coding gene nucleotide compositions, etc.), and predicted 22 transferRNA secondary structures. We obtained highly consistent phylogenetic results within Cicadellidae based on mitogenomic data using the maximum likelihood and Bayesian methods. Our results showed that all subfamilies were monophyletic and had a high node support rate, and there was a sister group relationship between Ledrinae and all other leafhopper groups. Furthermore, treehoppers were found to originate from leafhoppers and showed sister group relationships with Megophthalminae. Within Ledrinae, all phylogenetic trees supporting phylogenetic relationships were as follows: ([P. dicondylica + P. gongshanensis] + [Tituria pyramidata + [Ledra auditura + P. gongshanensis]]) Based on the complete mitogenome phylogenetic analysis and the comparison of morphological characteristics, we propose that Petalocephala is not monophyletic.

Comparative Mitochondrial Genome Analysis of Mamestra configurata (Lepidoptera: Noctuoidea: Noctuidae) and Other Noctuid Insects Reveals Conserved Genome Organization and Phylogeny

Abstract Mitochondrial genomes (mitogenomes) advance our understanding of molecular evolution and phylogenetic relationships. In this study, the mitogenome of Mamestra configurata was assembled, annotated, and analyzed. The total length of its genome was 15,322 bp; the A+T content accounted for 81.2%. Both the AT and GC skew values (−0.003 and −0.182) in the mitogenome of M. configurata were negative. Compared with their ancestral insects (trnI-trnQ-trnM), the gene sequence showed a typical trnm rearrangement (trnM-trnI-trnQ). Except for cox1, all the protein-coding genes were initiated with ATN. All tRNAs except trnS1 were of the four-leaf grass type. According to phylogenetic analysis, we found that M. configurata and Protegira songi clustered together in one clade with high node support values. Based on Bayesian inference and maximum likelihood methods, the phylogenetic results revealed that M. configurata was closely related to Protegira songi, the Erebidae was sister to (Nolidae + [Euteliidae + Noctuidae]), Doidae and Notodontidae is sister to the other families of Noctuoidea, respectively. Our results may provide a useful reference for the intensive study of the phylogeny of Noctuidae.

Evolving in the darkness: Phylogenomics of Sinocyclocheilus cavefishes highlights recent diversification and cryptic diversity

Troglomorphism—any morphological adaptation enabling life to the constant darkness of caves, such as loss of pigment, reduced eyesight or blindness, over-developed tactile and olfactory organs—has long intrigued biologists. However, inferring the proximate and ultimate mechanisms driving the evolution of troglomorphism (stygomorphism) in freshwater fish requires a sound understanding of the evolutionary relationships between surface and stygomorphic lineages. We use Restriction Site Associated DNA Sequencing (RADseq) to better understand the evolution of the Sinocyclocheilus fishes of China. With a remarkable array of derived stygomorphic traits, they comprise the largest cavefish diversification in the world, emerging as a multi-species model system to study evolutionary novelty. We sequenced a total of 120 individuals throughout the Sinocyclocheilus distribution. The data comprised a total of 646,497 bp per individual, including 4378 loci and 67,983 SNPs shared across a minimum of 114 individuals at a given locus. Phylogenetic analyses using either the concatenated RAD loci (RAxML) or the SNPs under a coalescent model (SVDquartets, SNAPP) showed a high degree of congruence with similar topologies and high node support (>95 for most nodes in the phylogeny). The major clades recovered conform to a pattern previously established using Sanger-based mt-DNA sequences, with a few notable exceptions. We now recognize six major clades in this group, elevating the blind cavefish S. tianlinensis and the micro-eyed S. microphthalmus as two new distinct clades due to their deep divergence from other clades. PCA plots of the SNP data also support the recognition of six major clusters of species congruent with the identified clades in ordination space. A Bayes factor delimitation (BFD) analysis showed support for 21 species, recognizing 19 previously described species and two putative new cryptic ones. Two species whose identities were previously disputed, S. furcodorsalis and S. tianeensis, are supported here as distinct species. In addition, our multi-species calibrated tree in SNAPP suggests that the genus Sinocyclocheilus originated around 10.16 Mya, with most speciation events occurring in the last 2 Mya, likely favored by the uplift of the Qinghai-Tibetan Plateau and cave occupation induced by climate-driven aridification during this period. These results provide a firm basis for future comparative studies on the evolution of Sinocyclocheilus and its adaptations to cave life.

Complete mitochondrial genome of Ricania shantungensis Chou & Lu, 1977 (Hemiptera: Ricaniidae)

Ricania shantungensis Chou & Lu, 1977 (Hemiptera: Ricaniidae), is an invasive pest that attacks forest as well as agricultural trees. We sequenced the 15,358 bp long complete mitochondrial genome (mitogenome) of this species; it consists of a typical set of genes (13 protein-coding genes, 2 rRNA genes, and 22 tRNA genes) and one major non-coding AT-rich region. The orientation and gene order of the R. shantungensis mitogenome are identical to that of the ancestral type found in majority of the insects. Bayesian inference (BI) phylogeny placed the R. shantungensis examined in our study, together with Ricania spp. in a group with the highest nodal support, forming the family Ricaniidae to which R. shantungensis belongs.

Two mitochondrial genomes of Taiwanese rhinoceros beetles, Oryctes rhinoceros and Eophileurus chinensis (Coleoptera: Scarabaeidae)

Two mitochondrial genomes of the dynastine beetles, Oryctes rhinoceros (Linnaeus, 1758) and Eophileurus chinensis (Faldermann, 1835), were assembled via high-throughput sequencing (HTS). Each of the mitogenomes has 37 genes, showing standard gene order and annotation as the other insects, except for the transfer genes, presenting tQ-tI-tM order. To examine their phylogenetic positions, 118 public mitogenomes of Scarabaeidae were used to infer a ML tree. Overall, our scarabaeid phylogeny reveals clear relationships with high nodal supports, and the two rhinoceros beetles are both grouped with the subfamily Dynastinae. The feeding habit of the two clades seems to represent coprophagous and phytophagous types. However, polyphyletic relationships were observed in the subfamily Melolothinae and in the tribes of Onthophagini and Oniticellini. Further systematic revision is needed.

The complete mitochondrial genome of Monopis longella Walker, 1863 (Lepidoptera: Tineidae)

The complete mitochondrial genome (mitogenome) of Monopis longella Walker, 1863 (Lepidoptera: Tineidae) comprises 15,541 bp and contains a typical set of genes and one non-coding region. The gene arrangement of M. longella is unique for Lepidoptera in that it has a trnI-trnM-trnQ sequence in the A + T-rich region and ND2 junction. Unlike most other lepidopteran insects, in which the COI gene has CGA as the start codon, M. longella COI has an ATT codon. Phylogenetic analyses based on the concatenated sequences of 13 protein-coding genes and two rRNA genes, using the Bayesian inference (BI) method, placed M. longella in the Tineidae, sister in position to the cofamilial species, Tineola bisselliella, with the highest nodal support. Tineidae, represented by three species including M. longella, formed a monophyletic group with high support (Bayesian posterior probability = 0.99). Within Tineoidea the sister relationship between Tineidae and Meessiidae was obtained with the highest support, leaving Psychidae occupying the basal lineage of the two families.

The complete mitogenome sequence of the hawk moth, Theretra latreillii subsp. lucasii (Lepidoptera: Sphingidae) from Zhejiang Province, China

The sphingid, Theretra latreillii subsp. lucasii is a common hawk moth distributed in southeast Asia and Australian regions. Although barcode analyses have been published, its complete mitogenome sequence has not been deciphered. In this study, the complete mitogenome of T. latreillii lucasii (GeneBank accession no. MW539688) was sequenced using Illumina HiSeq X Ten system for mitogenome-based phylogenetic analysis. The mitogenome was 15,354 bp in length and comprises 13 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, and 22 transfer RNAs (tRNAs) with the typical gene order and orientation of Sphingidae mitogenomes. The nucleotide composition of majority strand is 41.2% for A, 7.4% for G, 12.0% for C, and 39.4% for T, with an A + T content of 80.6%. Phylogenetic analysis using the 13 PCGs fully resolved T. latreillii lucasii in a clade with T. japonica, Macroglossum stellatarum, and Ampelophaga rubiginosa, with high nodal support both by Bayesian inference and maximum-likelihood methods, forming the Macroglossini monophyletic group.