Complete Mitogenome Research Articles

Sequencing and characterization of complete mitogenome DNA of worldwide turkey (Meleagris gallopavo) populations

The history of turkey (Meleagris gallopavo) domestication can be traced back to the period between 700 and 200 BC in Mexico. This process involved multiple contributors and resulted in the development of modern local turkey breeds. This research investigates the complete mitochondrial diversity across a diverse range of local turkeys. Seventy-three turkeys were sampled from various populations, including autochthonous Italian breeds, an American breed (Narragansett), as well as wild turkeys from the USA and Mexico. The mitochondrial DNA (mtDNA) was employed as a powerful tool for biodiversity and breed phylogeny investigation. An analysis of the entire mtDNA was conducted to identify breed-specific unique traits, mitochondrial-specific characteristics, and the phylogenetic relationship among turkey populations. A total of 44 polymorphic sites were identified. Brianzolo and Narragansett birds were characterized as genetically homogeneous populations. Thirty-two different haplotypes were identified when our samples were compared with mtDNA D-loop of 96 online available turkeys from various geographical countries. H1 and H2, differing for one mutation, were the most abundant, comprising 132 of the 185 sequences. H1 included samples coming from every region, while H2 was predominantly characterized by Italian samples. USA and Mexican samples appear to be more variable in their mtDNA than the other populations.

Integrative taxonomic revision of the grasshopper genera Parapetasia Bolívar, 1884, and Loveridgacris Rehn, 1954 (Orthoptera, Pyrgomorphidae), with description of a new species of Loveridgacris

The taxonomic status of the Pyrgomorphid genera Parapetasia Bolívar, 1884, and Loveridgacris Rehn, 1954 is complex and challenging. Here, we use a combination of morphological, distributional, and genetic data to revise the two genera and provide new information on their diversity. We describe a new species, Loveridgacris tectiferussp. nov., from Tanzania and formally resurrect the status of Parapetasia rammei as a valid species within Parapetasia, resulting in two species in Parapetasia (P. femorata and P. rammei) and two in Loveridgacris (L. impotens and L. tectiferussp. nov.). We also sequenced the COI and 16S genes of 10 Pyrgomorphidae species and provided the first phylogeny of the group. Our data show that all species are clearly distinct and represent molecular operational taxonomic units (mOTUs), with the exceptions of L. impotens and L. tectiferussp. nov., which are morphologically clearly distinct but for which the concatenated sequence alignments of the two individual gene datasets (COI and 16S) do not provide sufficient information. In addition, high interspecific distances were found between Parapetasia and Loveridgacris. Moreover, the complete mitogenomes of L. impotens and L. tectiferussp. nov. were sequenced using next-generation sequencing technology. The total lengths of the assembled mitogenomes were 15,592 bp and 15,737 bp, representing 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and one D-loop region, respectively. To aid in identification, we present a key for the two genera, including a key to species. This study provides insights into the morphology, distribution, and phylogeny of Pyrgomorphidae in Africa.

Complementing aculiferan mitogenomics: comparative characterization of mitochondrial genomes of Solenogastres (Mollusca, Aplacophora).

With the advances in high-throughput sequencing and bioinformatic pipelines, mitochondrial genomes have become increasingly popular for phylogenetic analyses across different clades of invertebrates. Despite the vast rise in available mitogenomic datasets of molluscs, one class of aplacophoran molluscs - Solenogastres (or Neomeniomorpha) - is still neglected. Here, we present six new mitochondrial genomes from five families of Solenogastres (Amphimeniidae, Gymnomeniidae, Proneomeniidae, Pruvotinidae, Simrothiellidae), including the first complete mitogenomes, thereby now representing three of the four traditional orders. Solenogaster mitogenomes are variable in size (ranging from approximately 15,000bp to over 17,000bp). The gene order of the 13 protein coding genes and two rRNA genes is conserved in three blocks, but considerable variation occurs in the order of the 22 tRNA genes. Based on phylogenetic analyses and reconstruction of ancestral mitochondrial genomes of Aculifera, the position of (1) trnD gene between atp8 and atp6, (2) trnT and P genes between atp6 and nad5, and (3) trnL1 gene between G and E, resulting in a 'MCYWQGL1E'-block of tRNA genes, are all three considered synapomorphies for Solenogastres. The tRNA gene block 'KARNI' present in Polyplacophora and several conchiferan taxa is dissolved in Solenogastres. Our study shows that mitogenomes are suitable to resolve the phylogenetic relationships among Aculifera and within Solenogastres, thus presenting a cost and time efficient compromise to approach evolutionary history in these clades.

Plio‐Pleistocene decline of mesic forest underpins diversification in a clade of Australian Panesthia cockroaches

AbstractThe progressive aridification of the Australian continent, and coincident decline of mesic forest, has been a powerful driver of allopatric and environmental speciation in native species. The relictual mesic forests of the eastern seaboard now harbour a diverse group of endemic fauna, including the wood‐feeding cockroaches of the genus Panesthia, which reached the continent via two separate invasions from Melanesia. The more recent of these colonization events gave rise to a group of five recognized species, occurring in mainland woodlands, sclerophylls and rainforests, as well as the forests and grasslands of the Lord Howe Island Group. Due to limited sampling in molecular studies and doubt regarding the standing taxonomy, there is little certainty about relationships among the species and poor understanding of the effects of ancient climatic change upon their evolution. We undertook a comprehensive phylogenetic analysis of the clade, using complete mitogenomes and nuclear ribosomal markers from nearly all known morphospecies and populations. Our time‐calibrated phylogenetic analyses reveal an additional six unrecognized, highly divergent lineages and suggest that these have arisen primarily through vicariance as rainforests fragmented during Plio‐Pleistocene glacial cycles (2–5 million years ago). Ancestral niche estimations also evidence a tropical rainforest origin for the group, followed by at least three niche transitions into drier forest, including one associated with the singular colonization of the Lord Howe Island Group. Finally, we find evidence of frequent, parallel wing reduction, in potential association with the contraction of forest habitats into small refugia. Our results reiterate the far‐reaching role of ancient aridification in driving speciation, niche expansion and morphological evolution in Australian fauna.

Three Mitochondrial Genomes of Chrysochroinae (Coleoptera, Buprestidae) and Phylogenetic Analyses

Three mitochondrial genomes of Chrysochroinae (Buprestidae) were sequenced and analyzed. The mitogenomes of the genera Catoxantha and Nipponobuprestis are first reportedand Chrysochroa opulenta is a first record for China. The complete mitogenomes of Catoxantha luodiana, Nipponobuprestis guangxiensis and Chrysochroa opulenta exhibit striking similarities in their lengths and composition. Specifically, their lengths are 15,594 bp, 15,775 bp and 15,587 bp, respectively. Each of these genomes encodes 37 typical mitochondrial genes. The overwhelming majority of protein-coding genes (PCGs) have the typical ATN (ATT, ATA, ATG or ATC) as the start codon and terminate with TAR (TAA or TAG) as the stop codon or an incomplete stop codon T-. Among the three mitogenomes, Leu2, Ser2 and Phe were the most frequently encoded amino acids. In the PCGs, the Ka/Ks ratio of cox1 is the lowest, whereas atp6 has the highest value. This suggests that cox1 can be used as a molecular barcode for species delimitation and phylogeny in Chrysochroinae. The phylogenetic results showed that C. luodiana and two Chrysochroa species formed a clade. Based on the topology of the phylogenetic tree, the genus Catoxantha should be reassigned as a subgenus of Chrysochroa.

The terrestrial flatworm Microplana scharffi (Geoplanidae, Microplaninae): mitochondrial genome, phylogenetic proximity to the Bipaliinae and genes related to regeneration

A genome skimming approach of sequencing was undertaken on a subfamily of terrestrial flatworms that had been neglected in genomic studies until now, namely the Microplaninae as represented here by Microplana scharffi. A single run of short-read sequencing enabled retrieval of the complete mitogenome, the two paralogous versions of the 18S gene, the elongation factor gene EF1α, plus two genes involved in the regeneration process, namely those coding for ß-CATENIN-1 and adenomatous polyposis coli (APC). The 15,297 bp mitogenome lacks a functional tRNA-Ala and has a mandatory alternative TTG start codon in its cox1 gene. The multiprotein phylogeny, inferred from mitogenome proteins, positions M. scharffi as sister-group to the Bipaliinae with maximum support, although the organisation of the mitogenomes shows features previously never observed among Bipaliinae, such as the conserved 32 bp overlap between ND4 and ND4L. Similarly to what has been observed in recent publications on other species of Geoplanidae, the two types of 18S genes display strongly different coverages and are only 90.57% identical. Additionally, alien DNA was identified in the pool of contigs in the form of the complete mitochondrial genome of Lumbricus rubellus, confirming previous observations on the feeding habits of M. scharffi.

Characterization of the complete mitochondrial genome of the Sunda stink-badger (Mydaus javanensis) from the island of Borneo.

The Mephitidae is a family of skunks and stink-badgers that includes 12 extant species in four genera, namely, Mydaus, Conepatus, Mephitis and Spilogale. Mydaus is the only genus within Mephitidae found outside the American continent, with its distribution limited to the islands of Borneo, Indonesia and Philippines. There are two extant species of Mydaus i.e., javanensis and marchei. Currently, complete mitogenomes are unavailable for either species. Here, we present the characterization of the first complete mitogenome for the Sunda stink-badger (Mydaus javanensis) from the island of Borneo. Muscle tissue was obtained and the DNA was sequenced using a combination of Illumina Barcode Tagged Sequence (BTSeq) and Sanger sequencing techniques. The genome was annotated with MITOS and manually checked for accuracy. A circular map of the mitogenome was constructed with Proksee. Relative synonymous codon usage (RSCU) and codon frequency were calculated using MEGA-X. The protein coding genes (PCGs) were aligned with reference sequences from GenBank and used for the construction of phylogenetic trees (maximum liklihood (ML) and Bayesian inference (BI)). Additionally, due to the lack of available complete genomes in public databases, we constructed another tree with the cyt b gene. The complete circular mitogenome was 16,391 base pairs in length. It comprises the typical 13 protein-coding genes, 22 tRNAs, two ribosomal RNA genes, one control region (CR) and an L-strand replication origin (OL). The G+C content was 38.1% with a clear bias towards A and T nucleotides. Of the 13 PGCs, only ND6 was positioned in the reverse direction, along with five other tRNAs. Five PCGs had incomplete stop codons and rely on post-transcriptional polyadenylation (TAA) for termination. Based on the codon count, Leucine was the most common amino acid (589), followed by Threonine (332) and Isoleucine (325). The ML and BI phylogenetic trees, based on concatenated PCGs and the cyt b gene, respectively, correctly clustered the species with other members of the Mephitidae family but were unique enough to set it apart from Conepatus, Mephitis and Spilogale. The results confirm Mydaus as a member of the mephitids and the mitogenome will be useful for evolutionary analysis and conservation of the species.

Genetic Insights into the Giant Keyhole Limpet (Megathura crenulata), an Eastern Pacific Coastal Endemic: Complete Mitogenome, Phylogenetics, Phylogeography, and Historical Demography

Background: The giant keyhole limpet Megathura crenulata is a gastropod mollusk (Fissurella superfamily) that is endemic to the eastern Pacific coast from southern California, USA, to Baja California Sur, Mexico. M. crenulata is socioeconomically important as it produces a potent immune-stimulating protein, called Keyhole Limpet Hemocyanin, which is extracted in vivo and utilized for vaccine development. However, ecological studies are scarce and genetic knowledge of the species needs to be improved. Our objectives were to assemble and annotate the mitogenome of M. crenulata, and to assess its phylogenetic relationships with other marine gastropods and to evaluate its population genetic diversity and structure. Methods: Samples were collected for mitogenome assembly (n = 3) spanning its geographic range, Puerto Canoas (PCA) and Punta Eugenia (PEU), Mexico, and California (CAL), USA. Total DNA was extracted from gills sequenced using Illumina paired-end 150-bp-read sequencing. Reads were cleaned, trimmed, assembled de novo, and annotated. In addition, 125 samples from eight locations were analyzed for genetic diversity and structure analysis at the 16s rRNA and COX1 genes. Results: The M. crenulata mitogenomes had lengths of 16,788 bp (PCA) and 16,787 bp (PEU) and were composed of 13 protein-coding regions, 22 tRNAs, two rRNAs, and the D-Loop region. In terms of phylogeographic diversity and structure, we found a panmictic population that has experienced recent demographic expansion with low nucleotide diversity (0.002), high haplotypic diversity (0.915), and low φST (0.047). Conclusions: Genetic insights into the giant keyhole limpet provides tools for its management and conservation by delimiting fishing regions with low genetic diversity and/or genetically discrete units.

Genomics of historical museum collections clarifies species diversity in Cuban hutias (Capromys)

Abstract Specimen-based taxonomic research is essential for understanding diversity and setting management frameworks for threatened mammal faunas, and ancient DNA techniques are increasingly used to extract information from taxonomically relevant historical specimens. The largest survivors of the depleted Caribbean mammal fauna are hutias in the genus Capromys, which is usually interpreted as containing a single species, C. pilorides. Previous studies have demonstrated genetic differentiation of Capromys populations across Cuba, but infrageneric species diversity and nomenclature remain unclear. We conducted ancient DNA analysis of historical Capromys samples using cytochrome b and complete mitogenome data sets, and including the 19th-century holotypes of 2 species now considered synonyms, C. fournieri and C. geayi. Our analyses identify distinct western and central/eastern Capromys clades that diverged 1.75 Mya based upon mitogenome data. These clades are separated by the Havana–Matanzas Channel, which represented a barrier to dispersal throughout the Neogene–Quaternary. Divergence date comparisons with other hutia species provide support for interpreting divergence between Capromys populations as species-level differentiation. Although we were unable to yield amplifiable DNA from the C. fournieri holotype, our analyses confidently assign the C. geayi holotype to the western Capromys clade. We therefore recognize 2 extant Capromys species: C. geayi (western Cuba) and C. pilorides (central/eastern Cuba and Cayman Islands).

Comparative analysis of mitochondrial genomes of invasive weed Mikania micrantha and its indigenous congener Mikania cordata

Mikania micrantha and Mikania cordata are two distinct species in China. The former is notorious as one of the top 100 worst invasive species, whereas the latter is an indigenous species harmless to native plants or the environment. They form an ideal congener pair for comparative studies aimed at deeply understanding the invasion mechanisms of the exotic weed. In this study, we have assembled and annotated the mitogenomes of both species using Illumina and PacBio sequencing data and compared their characteristic differences. The complete mitogenome of M. micrantha is a double-stranded DNA with a length of 336,564 bp, while the mitogenome of M. cordata exhibits a branching structure, consisting of two small circular molecules and six linear molecules, with a combined length totaling 335,444 bp. Compared to M. cordata, M. micrantha has less SSRs, tandem repeats, dispersed repeats, mitochondrial protein coding genes (PCGs). The two plants show similar codon usage patterns. This comparative study has revealed the structure and function of the mitogenomes of the two species and laid a solid foundation for investigating the effects of gene loss and duplication on the development of invasive traits in M. micrantha.

Mitogenomic Architecture of Atlantic Emperor Lethrinus atlanticus (Actinopterygii: Spariformes): Insights into the Lineage Diversification in Atlantic Ocean.

The evolutionary history of emperors, particularly in the Atlantic and Indo-West Pacific Oceans, remains largely unmapped. This study explores the maternal lineage evolution of Lethrinids by examining the complete mitogenome of Lethrinus atlanticus, which is endemic to the Eastern Atlantic Ocean. Utilizing advanced next-generation sequencing, we found that the mitogenome spans 16,789 base pairs and encompasses 37 genes, including 13 protein-coding genes (PCGs), two ribosomal RNAs, 22 transfer RNAs, and an AT-rich control region (CR). Our analysis indicates a preference for AT base pairs in the L. atlanticus mitogenome (53.10%). Most PCGs begin with the ATG codon, except for COI, which starts with GTG. Relative synonymous codon usage reveals high frequencies for alanine, leucine, proline, serine, and threonine. The ratio of nonsynonymous to synonymous substitutions suggests strong negative selection across all PCGs in Lethrinus species. Most transfer RNAs exhibit typical cloverleaf structures, with the exception of tRNA-serine (GCT), which lacks a dihydrouracil stem. Comparative analysis of conserved sequence blocks across the CRs of three Lethrinus species shows notable differences in length and nucleotide composition. Phylogenetic analysis using concatenated PCGs clearly distinguishes all Lethrinus species, including L. atlanticus, and sheds light on the evolutionary relationships among Spariformes species. The estimated divergence time of approximately 20.67 million years between L. atlanticus and its Indo-West Pacific relatives provides insights into their historical separation and colonization during the late Oligocene. The distribution of Lethrinids may be influenced by ocean currents and ecological factors, potentially leading to their speciation across the Eastern Atlantic and Indo-West Pacific. This study enhances our understanding of the genetic diversity and phylogenetic relationships within Lethrinus species. Further exploration of other emperor fish mitogenomes and comprehensive genomic data could provide vital insights into their genetic makeup, evolutionary history, and environmental adaptability in marine ecosystems globally.

The complete mitogenome of Alternaria tenuissima (Kunze) Wiltshire 1933 (Pleosporaceae), a fungus causing apple leaf blotch disease

Alternaria tenuissima (Kunze) Wiltshire 1933 is a plant pathogenic fungus mainly causing leaf blotch disease. Here, we de novo assembled mitochondrial genome of A. tenuissima isolate AT-1224. The total mitogenome size is 57,475 bp with 29.00% G + C content. The genome contained 12 coding genes and 15 hypothetical proteins, 34 transfer RNA (tRNA) genes and 2 ribosomal RNA (rRNA). There are 227 SSR repeats, range from 2 to 4 base pairs, most five repeats were AT (144), AAT (54), AG (33), AC (13) and AAG (5). The results also found 13 tandem repeats (>100 bp), the largest repeat were forward 2 times located from 13,405 to 20,024 bp and 25,549 to 32,168 bp. Phylogenetic analysis based on 17 species complete mitogenomes indicated that A. tenuissima mitogenome was closest to 2 species, A. solani and A. alternata, sister clade to 6 species, representing Curvularia clavate, Exserohilum rostratum, Exserohilum turcicum, Bipolaris cookie, Bipolaris oryzae and Bipolaris sorokiniana. Further analysis among common fungus in local apple orchards using mitochondrial protein-coding genes revealed A. tenuissima were closing to 2 Alternaria fungi and a fungus representing Phoma sp. These results provide a basic reference for identification and evolution studies of A. tenuissima on apple trees.

Complete mitochondrial genome of Saldoida armata Horváth, 1911 (Heteroptera: Saldidae) and phylogenetic analysis

The complete mitochondrial genome of Saldoida armata (Heteroptera: Saldidae) is 16,049 bp in length, comprising 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs) and a control region. All the PCGs are initially encoded by ATN, TTG or GTG, and terminated coding with TAA or a single T. With the exception of trnS(AGN), all tRNAs exhibit a typical cloverleaf secondary structure. Phylogenetic analysis reveals the sister relationship of S. armata with other Saldidae members. The complete mitogenome of S. armata will provide useful genetic information for species identification, phylogenetic analysis and conservation of this species.

Intraspecies mitogenomic variation in the Black-throated Tit (Aegithalos concinnus): conserved structure, concerted evolution of duplicate control regions and multiple distinct evolutionary lineages

The mitochondrial genome is a prominent research topic due to its indispensable role in organisms and its application in many research disciplines. However, few studies have investigated intraspecies mitogenomic variation. In this study, 69 mitogenomes of the Black-throated Tit (Aegithalos concinnus) were assembled and annotated from a large number of short reads generated using high-throughput sequencing technology. Comparative analyses revealed that mitogenomic characteristics such as length, gene and nucleotide composition, codon usage, and duplicated control regions were relatively conserved despite substantial intraspecies morphological changes. Yet, all the individuals from the subspecies A. c. iredalei had one more nucleotide in the 12S rRNA than the other studied subspecies. Phylogenetic analyses showed five distinct lineages based on the complete mitogenomes and the 13 combined protein-coding genes, whereas only four lineages were observed when using the duplicate control regions. Most interestingly, each lineage had both copies of the control regions of the comprising individuals, indicating that the paralogous control regions were more similar than the orthologous sequences from the distinct lineages. This suggested the control regions had undergone concerted evolution. The Black-throated Tit has complex evolutionary history and needs further investigating the taxonomic status of these lineages, as well as the underlying evolutionary processes. Our findings call for more research on intraspecies mitogenomic variation.

Compacted hair in broken teeth reveals dietary prey of historic lions.

With recent advances, nuclear genome data for phylogenomic analyses can now be sequenced from minuscule quantities of DNA1 and from specimens that are more than a million years old.2 DNA analysis from hair is a well-established approach3 widely used in forensic science4 and wildlife conservation.5 Hairsamples can be effectively decontaminated6 and can be used to identify the mammalian speciesfrom which the hair was shed.7,8 We aimed to use advances optimized for degraded DNA to systematically identify dietary prey species from hair compacted in the teeth of two Tsavo lions that livedduring the 1890s in Kenya (see description of samples in the STAR Methods and Patterson9 andKerbis Peterhans and Gnoske10 for background on the Tsavo "man-eaters"). Analysis of hair DNA identified giraffe, human, oryx, waterbuck, wildebeest, and zebra as prey and also identified hair thatoriginated from lion. DNA preservation allowed for analyses of complete mitogenome profiles of zebra, giraffe, and lion. Giraffe mitogenomes are phylogeographically partitioned, and we found thatthe lions ate at least two individuals that belong to a subspecies of Masai giraffe (Giraffa tippelskirchi tippelskirchi) typically found in southeast Kenya. The lion mitogenome from a hair sample was identical tothe Tsavo lion endogenous mitogenome and most closely matched other East African lions from Kenyaand Tanzania. Our approach enables a better understanding of the hunting behaviors, diets, and ecology of historical individuals, populations, and species and holds promise for extinct populations and species.

Complete mitochondrial genomes of two Phoretic mites (Parasitidae: Poecilochirus) on burying beetle: Genome characterization and phylogenetic analysis

A pair of intriguing phoretic instances are the burying beetle and Poecilochirus mites. Their interaction is environment-dependent and can be either mutualistic or antagonistic hostile depending on various biological and physical circumstances. In this study, we sequenced the complete mitogenomes of two Poecilochirus mite species, P. davydovae and P. mrciaki, which are associated with burying beetles, using Illumina HiSeq sequencing technology. Each mitochondrial genome comprised 37 typical genes, including 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and one control region (CR). The size of these mitogenomes were determined to be 14,489 bp and 14,517 bp, respectively. These two mitochondrial genomes contain a notable AT bias in their nucleotide composition, with positive AT and negative GC skews in the deflection index of mitochondrial genomes, and a unique incomplete stop codon T- in both protein-coding genes. The DHU arm of tRNA-S1 (AGN) only forms a simple loop structure, while the secondary structures of 21 tRNA genes in both species showed the conventional cloverleaf structures, like those of most metazoans. The phylogenetic tree of Gamasina was constructed using Maximum likelihood and Bayesian inference based on two rRNA genes and 13 PCGs. Based on the existing data, the Parasitidae forms a monophyletic, and Poecilochirus and Parasitus are closely related, possibly belonging to the same genus.

Comparative characterization and phylogenetic analysis of complete mitogenome of three taxonomic confused groupers and the insight to the novel gene tandem duplication in Epinephelus

Epinephelus bilobatus, Epinephelus maculatus and Epinephelus longispinis are three groupers that share common morphological characteristics and coloration patterns and have been morphologically confused and misidentified with each other for a long time. Complete mitochondrial genomes of the three groupers were determined and analyzed in this study. Mitogenomes of E. bilobatus, E. maculatus and E. longispinis were 17, 354 bp, 17, 066 bp and 17, 221 bp in size respectively and consisted of 13 protein-coding genes, two ribosomal RNA genes and one control region. However, different from most teleosts, which contain canonical 22 tRNAs, more numbers of tRNAs were identified in the three groupers with 27 tRNAs in E. bilobatus and E. longispinis and 25 tRNAs in E. maculatus. The increased number of tRNAs was due to the presence of tandemly duplicated tRNA-Asp genes that were located between tRNA-Ser and COII genes (six duplications in E. bilobatus and E. longispinis, four duplications in E. maculatus). Intact gene tandem duplication was an uncommon feature that was found in the typical teleost mitogenomes. The phylogenetic trees of the 32 groupers (genus Epinephelus) that were constructed based on 12 protein-coding genes revealed that Epinephelus species with tandemly duplicated tRNA-Asp genes were clustered into one monophyletic group, distinct from other Epinephelus species without any duplication features, which indicated that tandemly duplicated tRNA-Asp genes may be the particular linage-specific characteristics that evolve from a common ancestor and have the ability to distinguish them from other Epinephelus species. The results of the mitogenomes comparative analyses of the three groupers revealed the genetic distance of mitogenomes between each two species to be 0.062 (E. bilobatus vs E. maculatus), 0.091 (E. bilobatus vs E. longispinis) and 0.087 (E. maculatus vs E. longispinis). All values were far greater than the minimum value of 0.020 for species identification, which shows that they were three independent species at molecular level. Regarding the relationships between the three groupers, E. bilobatus was found to be more closely related to E. maculatus in comparison to E. longispinis. The results provide valuable molecular data for the species identification and phylogenetic analyses on E. bilobatus, E. maculatus and E. longispinis, and also provided a new insight into the tandem gene duplication features of Epinephelus mitogenomes.

Unraveling the Mitogenomic Characteristics and Phylogenetic Implications of Leuciscus merzbacheri (Zugmayer, 1912), an Endangered Fish in the Junggar Basin of Xinjiang, Northwest China.

Leuciscus merzbacheri is a rare and endangered fish in Xinjiang, China. As a representative species of the fauna in the Junggar Basin, it is of high economic and scientific value. The genetic data are still limited, and the mitochondrial genomic characteristics remain unexplored. A high-throughput sequencing method was used to obtain the complete mitogenome of L. merzbacheri. The full length of the circular DNA was 16,609 bp, and it consisted of 13 protein-coding genes (PCGs), 22 tRNAs, 2 rRNAs and 2 non-coding regions. The overall nucleotide compositions of both the mitogenome and PCGs showed an obvious AT preference with percentages of 54.20% and 53.60%, respectively. Three commonly used amino acids were Leu (16.43%), Ala (8.95%) and Thr (7.85%) in turn. All tRNAs could form the typical clover structures excluding tRNA-Ser AGY. The presumed secondary structures of two rRNAs contained several stem-loop domains, and the structure of 12S rRNA seemed to be more stable than that of 16S rRNA. Extended termination sequence regions (ETASs), central conserved regions (CSB-F, CSB-E and CSB-D), and conserved sequence regions (CSB-1, CSB-2 and CSB-3) were identified in the control region. The phylogenetic tree showed that L. merzbacheri was recovered with strong supports as a sister to the other members of the genus. The location in the outermost branch implied that it might be a relatively ancient species among its congeners. This study would complement the genetic data on L. merzbacheri and contribute to a better understanding of molecular evolution in Leuciscus as well.

Mitogenome-Based Phylogeny with Divergence Time Estimates Revealed the Presence of Cryptic Species within Heptageniidae (Insecta, Ephemeroptera).

Heptageniidae are known for their flat heads and bodies and are divided into three subfamilies. Despite the extensive diversity within this group and considerable efforts made to understand their evolutionary history, the internal classifications and origin time of Heptageniidae remains controversial. In this study, we newly sequenced 17 complete mitogenomes of Heptageniidae to reconstruct their phylogenetic positions within this family. Because of the ambiguous time of origin, our study also estimated the divergence time within Heptageniidae based on five fossil calibration points. The results of BI and ML trees all highly supported the monophyly of Heptageniidae and three subfamilies. The phylogenetic relationship of Rhithrogeninae + (Ecdyonurinae + Heptageniinae) was also recovered. The divergence time showed that Heptageniidae originated from 164.38 Mya (95% HPD, 150.23-181.53 Mya) in the mid-Jurassic, and Rhithrogeninae originated from 95.54 Mya (95% HPD, 73.86-120.19 Mya) in the mid-Cretaceous. Ecdyonurinae and Heptageniinae began to diverge at 90.08 Mya (95% HPD, 68.81-113.16 Mya) in the middle Cretaceous. After morphological identification, analysis of the mitogenome's composition, genetic distance calculation, phylogenetic analysis, and divergence time calculation, we suggest that two different populations of Epeorus montanus collected from Aksu, Xinjiang Uygur Autonomous Region (40°16' N, 80°26' E) and Xinyuan, Xinjiang Uygur Autonomous Region (43°20' N, 83°43' E) in China are cryptic species of E. montanus, but further detailed information on their morphological characteristics is needed to fully identify them.

Sampling poorly studied lineages improves resolution in mitogenome-based phylogenies of leafhoppers (Hemiptera: Cicadellidae)

Abstract Cicadellidae (leafhoppers) may be the most diverse and abundant insect herbivores in terrestrial ecosystems worldwide, with more than 23,000 described species in 20 extant subfamilies. Although prior studies have supported the monophyly of most recognized subfamilies, relationships among these groups remain inconsistently resolved and previous molecular phylogenies have failed to support the monophyly of a few subfamilies. To help address this deficiency, we combined 24 new complete mitogenomes representing 6 previously unsequenced subfamilies and 13 tribes with previously available data to create a dataset of 79 cicadellid taxa representing 16 subfamilies for phylogenetic analysis. The new analyses confirm that leafhopper mitochondrial genomes are highly conservative in overall structure, with only a few rare rearrangements of tRNAs. All phylogenetic analyses consistently supported the monophyly of previously recognized cicadellid tribes for which more than 1 representative was included and all of the included subfamilies were also consistently recovered as monophyletic with the exception of Aphrodinae and Eurymelinae. Bayesian inference and Maximum Likelihood analyses with site-homogeneous models are only slightly affected by nucleotide compositional heterogeneity and yield more stable phylogenetic relationships of family-group taxa than analyses with a heterogeneous-site mixture model using PhyloBayes. Molecular divergence time estimates indicate that the main lineages of Cicadellidae, most corresponding to recognized subfamilies, diverged during the Cretaceous period (93–121 Mya).