The complete mitochondrial genome of Eriocheir japonica sinensis (Decapoda: Varunidae) and its phylogenetic analysis

The complete mitochondrial genome (mitogenome) can provide novel insights into understanding the mechanisms underlying mitogenome evolution. In this study, the complete mitogenome of Eriocheir japonica sinensis (Decapoda: Varunidae) was determined to be 16,378 bp, including 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and a D-loop region. The AT skew of the E. j. sinensis mitogenome was slightly negative (−0.016), indicating a higher number of T compared with A nucleotides. The nucleotide composition of the mitogenome was also biased toward A + T nucleotides (71.6%). All PCGs were initiated by ATN codons. Eight of the 13 PCGs harbored the incomplete termination codon by T, or TA. All other tRNA genes displayed a typical clover-leaf structure of mitochondrial tRNA. The D-loop region of the E. j. sinensis mitogenome was 918 bp in length. Based on 13 PCGs, phylogenetic analysis confirmed the placement of E. j. sinensis within the Varunidae.     

The complete mitochondrial genome of Pardosa pusiola (Araneae,Lycosidae) and its phylogenetic implications

The mitochondrial genome (mitogenome) is useful for identification and phylogenetic analyses among arthropods, but there are no sufficient mitogenome data for wolf spiders. To enrich the mitogenome database of wolf spiders, the complete mitogenome of Pardosa pusiola was sequenced by high-throughput sequencing. It is 14,284 bp, comprising 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and a control region (CR). It represents a high bias toward A and T nucleotides with an A + T content of 76.49%. The mitogenome exhibited a negative AT skew (−0.13) and a positive GC skew (0.32). Most PCGs started with ATN codons and ended with TAA, TAG, or an incomplete T. In addition, most tRNAs had aberrant secondary structures with the absence of DHU arm or TΨC arm. Analysis performed with CREx software demonstrated that large-scale rearrangements of tRNAs were observed in the mitogenome of P. pusiola as compared with the putative ancestral mitogenome. The Bayesian inference (BI) and maximum likelihood (ML) phylogenetic trees based on the 13 PCGs of 25 spiders had the same topology, which could be presented as (Araneidae + (Agelenidae + (Dictynidae + Desidae)) + (Salticidae + (Thomisidae + (Oxyopidae + (Pisauridae + Lycosidae))))). This study offers a useful genetic resource for the taxonomy and phylogeny of spiders.     

Characterization of the complete mitochondrial genome of Phodopus roborovskii (Rodentia: Cricetidae) and systematic implications for Cricetinae phylogenetics

Phodopus roborovskii (subfamily Cricetinae) is widely distributed in the northern arid regions of China. This study reports its complete mitochondrial genome (mitogenome) for the first time. The complete sequence was 16,273 bp long, including 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and 1 major noncoding region. The base composition and codon usage were described. The putative origin of replication for the light strand (O_L) of P. roborovskii was approximately 45 bp long and was highly conserved in the stem-loop and adjacent sequences, but the starting sequence of replication varied between genera among Rodentia. We analyzed the three domains of the D-loop region, and the results indicated that the central domain had higher G + C content and lower A + T content than two peripheral domains. Phylogenetic analyses indicated high resolution in four main divergent clades using mitogenomes data within Cricetidae. Within Cricetinae clade, P. roborovskii was at basal position which was in line with previous researches, and it shared a common ancestor with other extant hamsters. This work validated previous molecular and karyotype researches using mitogenomes data, and provided a set of useful data on phylogeny and molecular evolution in Cricetidae species.     

The complete mitochondrial genome of Parasesarma pictum (Brachyura: Grapsoidea: Sesarmidae) and comparison with other Brachyuran crabs

Mitochondrial DNA (mtDNA) is an extrachromosomal genome which can provide important information for evolution and phylogenetic analysis. In this study, we assembled a complete mitogenome of a crab Parasesarma pictum (Brachyura: Grapsoidea: Sesarmidae) from next generation sequencing reads at the first time. P. pictum is a mudflat crab, belonging to the Sesarmidae family (subfamily Sesarminae), which is perched on East Asia. The 15,716 bp mitogenome covers 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and one control region (CR). The control region spanns 420 bp. The genome composition was highly A+T biased 75.60% and showed negative AT-skew (−0.03) and negative GC-skew (−0.47). Compared with the ancestor of Brachyura, the gene order of Sesarmidae has several differences and the gene order of P. pictum is typical for mitogenomes of Sesarmidae. Phylogenetic tree based on nucleotide sequences of mitochondrial 13 PCGs using BI and ML determined that P. pictum has a sister group relationship with Parasesarma tripectinis and belongs to Sesarmidae.     

Complete mitochondrial genome of the stonefly Cryptoperla stilifera Sivec (Plecoptera: Peltoperlidae) and the phylogeny of Polyneopteran insects

We present the complete mitogenome of a stonefly, Cryptoperla stilifera Sivec (Plecoptera; Peltoperlidae). The mitogenome was a circular molecule consisting of 15,633 nucleotides, 37 genes and a A + T-rich region. C. stilifera mitogenome was similar to Pteronarcys princeps mitogenome (Plecoptera; Pteronarcyidae). All transfer RNA genes (tRNAs) had typical cloverleaf secondary structures except for trnSer (AGN), where the stem-loop structure of the dihydrouridine (DHU) arm was missing. The A + T-rich region of C. stilifera had two stem-loops and each had two interlink. Three conserved sequence blocks (CSBs) were present in the A + T-rich regions of C. stilifera, Peltoperla tarteri and Peltoperla arcuata. Moreover, many polynucleotide stretches (Poly N, N = A, T and C) in the A + T-rich region of C. stilifera Phylogenetic relationships of Polyneopteran species were constructed based on the nucleotide sequences of 13 protein coding genes (PCGs). Both maximum likelihood (ML) and Bayesian inference (BI) analyses supported Grylloblattodea as the sister group to Plecoptera + Dermaptera and Embiidina and Phasmatodea as sister groups.     

The complete mitochondrial genome sequence of the cutlassfish Trichiurus japonicus (Perciformes: Trichiuridae): Genome characterization and phylogenetic considerations

Mitochondrial genome sequence and structure analysis has become a powerful tool for studying molecular evolution and phylogenetic relationships. To understand the systematic status of Trichiurus japonicus in suborder Scombroidei, we determined the complete mitochondrial genome (mitogenome) sequence using the long-polymerase chain reaction (long-PCR) and shotgun sequencing method. The entire mitogenome is 16,796 bp in length and has three unusual features, including (1) the absence of tRNA^Pro gene, (2) the possibly nonfunctional light-strand replication origin (O_L) showing a shorter loop in secondary structure and no conserved motif (5'-GCCGG-3'), (3) two sets of the tandem repeats at the 5' and 3' ends of the control region. The three features seem common for Trichiurus mitogenomes, as we have confirmed them in other three T. japonicus individuals and in T. nanhaiensis. Phylogenetic analysis does not support the monophyly of Trichiuridae, which is against the morphological result. T. japonicus is most closely related to those species of family Scombridae; they in turn have a sister relationship with Perciformes members including suborders Acanthuroidei, Caproidei, Notothenioidei, Zoarcoidei, Trachinoidei, and some species of Labroidei, based on the current dataset of complete mitogenome. T. japonicus together with T. brevis, T. lepturus and Aphanopus carbo form a clade distinct from Lepidopus caudatus in terms of the complete Cyt b sequences. T. japonicus mitogenome, as the first discovered complete mitogenome of Trichiuridae, should provide important information on both genomics and phylogenetics of Trichiuridae.     

The complete mitochondrial genome of Plesiastrea versipora (Scleractinia,Plesiastreidae) sheds light on its phylogeny and taxonomy of the family Plesiastreidae

The genus Plesiastrea used to be a member of the traditional family Faviidae, falling into the challenging ‘Bigmessidae’ clade, and was re-established until recent molecular phylogenies published. The entire mitogenome of the symbiotic coral Plesiastrea versipora (Lamarck, 1816), the type species of the family Plesiastreidae, was sequenced. The length of the mitochondrial genome is 15,320 bp and it includes thirteen protein-coding genes (PCGs), two rRNAs and two tRNAs. The nucleotide composition of GC is 32%. We perform phylogenetic reconstruction based on maximum likelihood (ML) and Bayesian analysis(BI) using all PCGs. Our result indicates that P. versipora clusters closely with species which belong to Mussidae, Merulinidae and Lobophylliidae. Our phylogenetic analyses provide solid evidence for phylogenetic placement of P. versipora and the evolutionary relationships among different families within the traditional robust clade of Scleractinia. In addition, the mitogenome data provide useful information for further molecular systematic investigations on Plesiastreidae as well as conservation biology research of P. versipora.     

Novel gene rearrangement in the mitochondrial genome of Coenobita brevimanus (Anomura: Coenobitidae) and phylogenetic implications for Anomura

The complete mitochondrial genomes (mitogenomes) can indicate phylogenetic relationships among organisms, as well as useful information about the process of molecular evolution and gene rearrangement mechanisms. However, knowledge on the complete mitogenome of Coenobitidae (Decapoda: Anomura) is quite scarce. Here, we describe in detail the complete mitogenome of Coenobita brevimanus, which is 16,393 bp in length, and contains 13 protein-coding genes, two ribosomal RNA, 22 transfer RNA genes, as well as a putative control region. The genome composition shows a moderate A + T bias (65.0%), and exhibited a negative AT-skew (−0.148) and a positive GC-skew (0.183). Five gene clusters (or genes) involving eleven tRNAs and two PCGs were found to have rearranged with respect to the pancrustacean ground pattern gene order. Duplication-random loss and recombination models were determined as most likely to explain the observed large-scale gene rearrangements. Phylogenetic analysis placed all Coenobitidae species into one clade. The polyphyly of Paguroidea was well supported, whereas the non-monophyly of Galatheoidea was inconsistence with previous findings on Anomura. Taken together, our results help to better understand gene rearrangement process and the evolutionary status of C. brevimanus and lay a foundation for further phylogenetic studies of Anomura.     

Characterization of the complete mitogenome of Trachylophus sinensis (Coleoptera: Cerambycidae: Cerambycinae), the type species of Trachylophus and its phylogenetic implications

Complete mitochondrial genomes (mitogenomes) have long been proved as reliable markers for phylogenetic reconstruction among diverse animal groups, especially benefited from recent rapid development of sequencing techniques. However, the mitogenomes of many important clades remain poorly represented, which restricted the understanding of macroscale evolutionary history of these groups. Here, we sequenced and characterized the complete mitogenome of Trachylophus sinensis, a type species of the Trachylophus genus, which also represents the first sequenced mitogenome in this genus. The complete circular mitogenome was 15,746 bp in length, containing 37 typical genes and one noncoding AT-rich control region. The nucleotide composition of the mitogenome was highly A + T biased, accounting for 70.07 % of the whole mitogenome with a slightly positive AT skewness (0.106). The 13 Protein coding genes (PCGs) used ATN as their start codons, except nad1 which used TTG. All tRNA genes were predicted with a characteristic cloverleaf secondary structure except trnS1^(AGN), whose dihydrouridine (DHU) arm was replaced by a simple loop. Phylogenetic analyses recovered Cerambycinae as a monophyletic group with high node supports and the sister relationship between T. sinensis and Nadezhdiella cantori. However, we found that deeper nodes showed not strong support, which may be caused by limited taxa sampling in our study. More mitogenomes should be sequenced representing various taxonomic levels, especially closely related species, which will enhance our understanding of phylogenetic relationships among Cerambycinae.     

The complete mitochondrial genome of Nephrotoma scalaris parvinotata (Brunetti 1918) (Diptera: Tipulidae) and insights into the phylogeny of Tipulomorpha

Craneflies (Tipuloidea or Tipulidae sensu lato) are one of the most diverse groups of true flies (Insecta, Diptera). The Tipuloidea and perhaps the Trichoceridae formed the infraorder Tipulomorpha, which is traditionally considered the most basal group of Diptera. Relationships among Tipulomorpha and the phylogenetic position of this infraorder within the whole Diptera remain to be settled. A mitochondrial genome (mitogenome) phylogeny of Diptera was produced to test the relationships within Tipulomorpha and its phylogenetic position. A complete mitogenome of Nephrotoma scalaris parvinotata (Tipuloidea, Tipulidae) was firstly sequenced with a next-generation sequencing approach. Compared with the published mitogenomes of Tipuloidea, the new mitogenome had a larger genome size (17,862 nt), due to a longer non-coding control region. The gene order was identical to Drosophila yakuba. Phylogenetic reconstructions using different inference methods recovered Tipulomorpha as monophyletic. And the Tipulomorpha was retrieved in a relatively basal position in Diptera. Within Tipulomorpha, the Tipuloidea and the Trichoceridae were strongly supported as reciprocally monophyletic. Relationships within Tipuloidea were resolved as (Pedicidae + (Limoniidae + (Cylindrotomidae + Tipulidae))). Well supported relationships include: Pedicidae was the sister group of (Limoniidae + (Tipulidae + Cylindrotomidae)); Limoniidae was paraphyletic with respect to (Tipulidae + Cylindrotomidae); Cylindrotomidae was the sister group of Tipulidae. The newly sequenced N. scalaris parvinotata clustered with other two Nephrotoma species at a derived position in Tipuloidea.     

The complete mitochondrial genome of Diaphorina citri (Hemiptera: Psyllidae) and phylogenetic analysis

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is the most serious pest of citrus as the vector of Huanglongbing (HLB), the citrus greening disease. In this study, the complete mitochondrial genome (mitogenome) of D. citri has been sequenced and annotated, and a comparative analysis is provided with known Psylloidea species. The mitogenome of D. citri is a typical circular molecular of 15,038 bp in length with an A + T content of 74.56%, contains the typical 37 genes and the gene order is identical to the other Psylloidea mitogenomes. The nucleotide composition and codon usage of D. citri are similar to the four Psylloidea species. All protein-coding genes (PCGs) use standard initiation codons (TAN), stop with TAA and TAG except ND2 and ND5 which stop with incomplete termination codon T. All tRNAs have the typical clover-leaf structure, with the exception of trnS1 lacking the dihydrouridine (DHU) arm. The control region is located between rrnL and the trnI gene with the highest A + T content among the five Psylloidea species. Phylogenetic analysis is conducted based on the 13 PCGs or/and 2 rRNAs of 23 Sternorrhycha mitogenomes. Both maximum likelihood (ML) and Bayesian inference (BI) analysis suggest a clear relationship of Psylloidea, Aleyrodoidea and Aphidoidea within Sternorrhycha, which support the traditional morphological classification.     

Two mitogenomes in Gruiformes (<Emphasis Type="Italic">Amaurornis akool</Emphasis>/<Emphasis Type="Italic">A. phoenicurus</Emphasis>) and the phylogenetic placement of Rallidae

Rallidae, with 34 genera including 142 species, is the largest family in the Gruiformes, the phylogenetic placement of this family was still in debate. The complete mitochondrial genomes (mitogenomes), with many advantageous characters, have become popular markers in phylogenetic analyses. We sequenced the mitogenomes of brown crake (Amaurornis akool) and white-breasted waterhen (Amaurornis phoenicurus), analyzed the genomic characters of mitogenomes in Rallidae, and explored the phylogenetic relationships between Rallidae and other four families in Gruiformes based on mitogenome sequences of 32 species with Bayesian method. The mitogenome of A. akool/A. phoenicurus was 16,950/17,213 bp in length, and contained 37 genes typical to avian mitogenomes and one control region, respectively. The genomic characters of mitogenomes in Rallidae were similar. The phylogenetic results indicated that, among five families, Rallidae had closest relationship with Heliornithidae, which formed a sister taxa to Gruidae, while Rhynochetidae located in the basal lineage. Within Rallidae, Rallina was ancestral clade. Gallirallus & Rallus and Aramides were closely related, Gallicrex & Amaurornis and Fulica & Gallinula had close relationships, and these two taxa formed a sister clade to Porphyrio & Coturnicops. Our phylogenetic analyses provided solid evidence for the phylogenetic placement of Rallidae and the evolutionary relationships among different genus within this family. In addition, the mitogenome data presented here provide useful information for further molecular systematic investigations on Gruiformes as well as conservation biology research of these species.     

Complete mitochondrial genome sequence of the Himalayan Griffon,Gyps himalayensis (Accipitriformes: Accipitridae): Sequence,structure, and phylogenetic analyses

This is the first study to describe the mitochondrial genome of the Himalayan Griffon, Gyps himalayensis, which is an Old World vulture belonging to the family Accipitridae and occurring along the Himalayas and the adjoining Tibetan Plateau. Its mitogenome is a closed circular molecule 17,381 bp in size containing 13 protein‐coding genes, 22 tRNA coding genes, two rRNA‐coding genes, a control region (CR), and an extra pseudo‐control region (CCR) that are conserved in most Accipitridae mitogenomes. The overall base composition of the G. himalayensis mitogenome is 24.55% A, 29.49% T, 31.59% C, and 14.37% G, which is typical for bird mitochondrial genomes. The alignment of the Accipitridae species control regions showed high levels of genetic variation and abundant AT content. At the 5′ end of the domain I region, a long continuous poly‐C sequence was found. Two tandem repeats were found in the pseudo‐control regions. Phylogenetic analysis with Bayesian inference and maximum likelihood based on 13 protein‐coding genes indicated that the relationships at the family level were (Falconidae + (Cathartidae + (Sagittariidae + (Accipitridae + Pandionidae))). In the Accipitridae clade, G. himalayensis is more closely related to Aegypius monachus than to Spilornis cheela. The complete mitogenome of G. himalayensis provides a potentially useful resource for further exploration of the taxonomic status and phylogenetic history of Gyps species.     

The complete mitochondrial genome of the sycamore lace bug Corythucha ciliata (Hemiptera: Tingidae)

The complete mitochondrial genome of the sycamore lace bug, Corythucha ciliata, was sequenced in this study. It represents the first sequenced mitogenome of family Tingidae in Heteroptera. The mitogenome of C. ciliata is 15,257 bp and contains 37 genes including 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes and a large non-coding region. Gene arrangement, nucleotide content, codon usage, and amino acid composition and asymmetry indicate a high degree of conservation with six other species of Cimicomorpha. The 13 PCGs initiated with ATN as the start codon and terminated with TAA, TA or T as stop codon. The evolutionary rate of each PCG was different, among which ATP8 showed the highest rate while ATP6 indicated the lowest rate. The 22 tRNAs genes apparently fold into a typical cloverleaf structure; however, the anticodon (TTC) of trnSer (AGN) differs from other Heteropteran insects. Secondary structure modeling of rRNA genes revealed similarity to other insects, except for two incomplete helices (H1648 and H2735) in lrRNA. The predicted secondary structure of lrRNA indicates 45 helices in six domains, whereas srRNA has 27 helices in three domains. Three potential stem–loops and two tandem repeats (–TCTAAT–) were identified in the A+T-rich region. Phylogenetic analysis indicated that C. ciliata is a sister group to other Heteroptera species based on analysis of the 13 PCGs.     

Complete mitochondrial genome of the forensically important carrion beetle,Necrodes nigricornis (Coleoptera: Silphidae)

The carrion beetle, Necrodes nigricornis Harold, 1875 (Coleoptera: Silphidae) is one of the most frequently encountered insect species among Silphidae in animal cadavers in Korea. Consequently, the postmortem colonization of the species can provide useful information regarding the time-since-death. In this study, we report the mitochondrial genome (mitogenome) characteristics of N. nigricornis with the aim of increasing the genomic data pools of the family Silphidae, which is of considerable forensic importance. The full-length circular genome is 18,503 bp, with 75.60 % A/T content. It contains a typical set of 37 metazoan genes, but the non-coding A + T-rich region is unusually long at 3,818 bp in length, containing two similar-sized repeats (539 and 542 bp) with a slight length and nucleotide variation. The gene arrangement of the species is identical to that of the ancestral arrangement found in the majority of insects. The biased A/T content in the genome is also reflected in the form of biased codon usage; six codons among 62 comprised solely of A/T nucleotides (TTA, ATT, TTT, ATA, TTA, and TAT) were highly used, accounting for 41.28 % of the total. The maximum-likelihood tree built using 12 PCG (excluding ND2) and lrRNA revealed monophyletic Silphidae, Silphinae, and Nicrophorinae, with relatively higher nodal supports (bootstrap support ≥ 80 %). N. nigricornis, belonging to Silphinae, was placed as the sister to congeneric N. littoralis with the highest nodal support. We believe this mitogenome sequences of the carrion beetle can become an important baseline information for future studies on phylogeny at various levels of taxonomic diversity, intra-specific variation, as well as species identification.     

The Entire Mitochondrial Genome of Macrophthalmus abbreviatus Reveals Insights into the Phylogeny and Gene Rearrangements of Brachyura

Brachyuran crabs comprise the most species-rich clades among extant Decapoda and are divided into several major superfamilies. However, the phylogeny of Brachyuran remains controversial, comprehensive analysis of the overall phylogeny is still lacking. Complete mitochondrial genome (mitogenome) can indicate phylogenetic relationships, as well as useful information for gene rearrangement mechanisms and molecular evolution. In this study, we firstly sequenced and annotated the complete mitogenome of Macrophthalmus abbreviatus (Brachyura; Macrophthalmidae). The mitogenome length of M. abbreviatus is 16,322 bp, containing the entire set of 37 genes and a control region typically observed in Brachyuran mitogenomes. The genome composition of M. abbreviatus was highly A+T biased 76.3% showing positive AT-skew (0.033) and negative GC-skew (??0.351). In M. abbreviatus mitogenome, most tRNA genes were folded into the clover-leaf secondary structure except trnH, trnS1 and trnC, which was similar to the other species in Macrophthalmidae. Phylogenetic analysis showed that all families form a monophyletic, and Varunidae and Macrophthalmidae clustered into a monophyletic clade as sister groups. Comparative analyses of rearrangement among Brachyura revealed that Varunidae (Grapsoidea) and Macrophthalmidae (Ocypodoidea) had the same gene order, which reinforced the result of phylogeny. The combined results of two aspects revealed that the polyphyly of Ocypodoidea and Grapsoidea were well supported. In general, the results obtained in this research will contribute to further studies on molecular based for the classification and gene rearrangements of Macrophthalmidae or even Brachyura.

     

The complete mitogenome of Bombyx mori strain Dazao (Lepidoptera: Bombycidae) and comparison with other lepidopteran insects

The complete mitochondrial genome (mitogenome) of Bombyx mori strain Dazao (Lepidoptera: Bombycidae) was determined to be 15,653 bp, including 13 protein-coding genes (PCGs), two rRNA genes, 22 tRNA genes and a A + T-rich region. It has the typical gene organization and order of mitogenomes from lepidopteran insects. The AT skew of this mitogenome was slightly positive and the nucleotide composition was also biased toward A + T nucleotides (81.31%). All PCGs were initiated by ATN codons, except for cytochrome c oxidase subunit 1 (cox1) gene which was initiated by CGA. The cox1 and cox2 genes had incomplete stop codons consisting of just a T. All the tRNA genes displayed a typical clover-leaf structure of mitochondrial tRNA. The A + T-rich region of the mitogenome was 495 bp in length and consisted of several features common to the lepidopteras. Phylogenetic analysis showed that the B. mori Dazao was close to Bombycidae.     

The mitogenome of freshwater loach Homatula laxiclathra (Teleostei: Nemacheilidae) with phylogenetic analysis of Nemacheilidae

The complete mitogenome can provide valuable genetic information to reconstruct relationships between species. In this study, we sequenced a stone loach, Homatula laxiclathra (Teleostei: Nemacheilidae), which is found in the northern region of the Qinling Mountains in China. The size of the H. laxiclathra mitogenome is 16,570 bp, which contains 37 typical mitochondrial genes including 13 protein‐coding genes, 22 transfer RNAs, two ribosomal RNAs, and a control region (D‐loop) with a total AT content of 55.8%. This is similar to other Nemacheilidae sequences published in GenBank. Furthermore, a mito‐phylogenomic analysis of 46 Nemacheilidae species places H. laxiclathra in a robust monophyletic Homatula cluster with other Homatula species. Our results contribute toward a better understanding of a true phylogeny of these species based on large‐scale taxonomic samplings as well as to help grasp the evolution of fish mitogenomes.     

The enigmatic Crimean green lizard (Lacerta viridis magnifica) is extinct but not valid: Mitogenomics of a 120-year-old museum specimen reveals historical introduction

It has been proposed that Lacerta viridis magnifica Sobolevssky, 1930 represents an extinct species or subspecies of green lizard endemic to the southern Crimea. Using NGS protocols optimized for heavily degraded DNA, we sequenced the complete mitogenome of one of the originally formalin-preserved specimens collected in the late 19th century. A comparison with sequence data of other green lizards revealed that L. v. magnifica is a junior synonym of the northern subspecies of the western green lizard (L. b. bilineata Daudin, 1802), which occurs at least 1,500 km away, beyond the distribution ranges of other green lizards. In medieval times, a Genoese colony existed in the Crimean region where the extinct green lizards occurred. Until the early 20th century, close ties to Italy persisted, and locals of Genoese descent sent their children for education to Italy, where L. b. bilineata occurs. This suggests that the extinct Crimean green lizards have been introduced accidentally or intentionally from Italy. Our study exemplifies the value of historical formalin-preserved museum specimens for clarifying the status of questionable rare or extinct taxa.