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Simple Summary: The spot-legged treefrog Polypedates megacephalus (Anura: Rhacophoridae) is widely distributed in Asia. In previous studies, it has been noted that the mitochondrial gene ATP8 has not been confidently annotated in any sequenced Polypedates to date. Duplications of control regions (CRs) are often observed in the tree frog family Rhacophoridae, and in most cases the copied CRs are highly similar to one another. This feature may lead to sequence assembly errors. Our findings are the first to detect two copies of the ND5 genes and three copies of the CRs in P. megacephalus by employing a "primer bridging" approach and allude to the presence of ATP8 via bioinformatic analyses and RT-qPCR. However, the question of whether the ATP8 is functional needs to be addressed in future studies. In prior research on the mitochondrial genome (mitogenome) of Polypedates megacephalus, the one copy of ND5 gene was translocated to the control region (CR) and the ATP8 gene was not found. Gene loss is uncommon among vertebrates. However, in this study, we resequenced the mitogenomes of P. megacephalus from different regions using a "primer bridging" approach with Sanger sequencing technologies, which revealed the "missing" ATP8 gene in P. megacephalus as well as three other previously published Polypedates. The mitogenome of this species was found to contain two copies of the ND5 genes and three copies of the control regions. Furthermore, multiple tandem repeats were identified in the control regions. Notably, we observed that there was no correlation between genetic divergence and geographic distance. However, using the mitogenome, gene expression analysis was performed via RT-qPCR of liver samples and it was thus determined that COIII, ND2, ND4, and ND6 were reduced to 0.64 ± 0.24, 0.55 ± 0.34, 0.44 ± 0.21 and 0.65 ± 0.17, respectively, under low-temperature stress (8 °C) as compared with controls (p < 0.05). Remarkably, the transcript of long non-coding RNA (lncRNA) between positions 8029 and 8612 decreased significantly with exposure to low-temperature stress (8 °C). Antisense ND6 gene expression showed a downward trend, but this was not significant. These results reveal that modulations of protein-coding mitochondrial genes and lncRNAs of P. megacephalus play a crucial role in the molecular response to cold stress. [ABSTRACT FROM AUTHOR]

Simple Summary: Hylidae is a rich family of Anura that is widely distributed across the world. Previous studies have shown that the Hyla and Dryophytos genera in the Hylidae used to belong to the same genus. The Eurasian species (including Hyla sanchiangensis) originated in North America and spread through the Beringian Land Bridge to Asia during the last ice age. Adaptation to low temperatures may require more energy expenditure. Mitochondria are the center of energy metabolism and, through oxidative phosphorylation, provide most of the ATP energy for the physiological and biochemical activities of the body. The mitogenome was used to investigate whether Hyla and Dryophytos genera are subject to positive selection. In addition, as a unique species in China, the distinction between Hyla sanchiangensis (Anura: Hylidae) and two different sites was also compared and analyzed in this paper. Hyla sanchiangensis (Anura: Hylidae) is endemic to China and is distributed across Anhui, Zhejiang, Fujian, Guangdong, Guangxi, Hunan, and Guizhou provinces. The mitogenomes of H. sanchiangensis from two different sites (Jinxiu, Guangxi, and Wencheng, Zhejiang) were sequenced. Phylogenetic analyses were conducted, including 38 mitogenomes of Hylidae from the NCBI database, and assessed the phylogenetic relationship of H. sanchiangensis within the analyzed dataset. Two mitogenomes of H. sanchiangensis showed the typical mitochondrial gene arrangement with 13 protein-coding genes (PCGs), two ribosomal RNA genes (12S rRNA and 16S rRNA), 22 transfer RNA (tRNA) genes, and one non-coding control region (D-loop). The lengths of the 12S rRNA and 16S rRNA genes from both samples (Jinxiu and Wencheng) were 933 bp and 1604 bp, respectively. The genetic distance (p-distance transformed into percent) on the basis of the mitogenomes (excluding the control region) of the two samples was calculated as 4.4%. Hyla sanchiangensis showed a close phylogenetic relationship with the clade of (H. annectans + H. tsinlingensis), which was supported by ML and BI analyses. In the branch-site model, five positive selection sites were found in the clade of Hyla and Dryophytes: Cytb protein (at position 316), ND3 protein (at position 85), and ND5 protein (at position 400) have one site, respectively, and two sites in ND4 protein (at positions 47 and 200). Based on the results, we hypothesized that the positive selection of Hyla and Dryophytes was due to their experience of cold stress in historical events, but more evidence is needed to support this conclusion. [ABSTRACT FROM AUTHOR]

Simple Summary: The sequencing of mitochondrial genomes promotes the study of cryptic species and the phylogenetic relationship of species. Thus far, only one complete mitochondrial genome of Scutigeromorpha is available in the NCBI. In this study, four specimens of Thereuonema tuberculata (Scutigeromorpha: Scutigeridae), collected from four different localities of China, were identified, and the four mitochondrial genomes of those were sequenced and annotated. Based on the gene organization and genetic diversity of the mitochondrial genomes, we hypothesized that cryptic species could exist in T. tuberculata. We further constructed BI and ML phylogenetic trees to reveal the relationship of Scutigeromorpha. Based on morphological characteristics to make species identification, the cryptic species of the Scutigeromorpha can be greatly underestimated. The mitochondrial genome provides a desirable tool for the biological identifications and the discovery of the cryptic species. The capacity to acquire mitochondrial genome sequences has substantially improved in recent years using next-generation sequencing (NGS) technology. On the basis of the next-generation sequencing, we obtained four complete mitochondrial genomes of Thereuonema tuberculata (Wood, 1862) from Nanyang, Henan Province (NY), Nanchang, Jiangxi Province (NC), Jinan, Shandong Province (JN), and Dali, Yunnan Province (DL) in China with GenBank numbers OK513221, OL449685, ON058988 and ON058989, respectively. The lengths of the four mitochondrial genomes ranged from 14,903 to 14,909 bp. The composition and order of genes of the four mitochondrial genomes were identical to the published mitochondrial genome of Scutigera coleoptrata (Linnaeus, 1758) (Scutigeromorpha: Scutigerdae). It was the first time that the tandem repeats in the control region were detected in Scutigeromorpha. We also calculated the corrected pairwise genetic distance of four complete mitochondrial genomes of T. tuberculata, ranging from 7.7 to 15.2%. The results showed that the T.tuberculata NC belonged to the typical sample of T. tuberculata, and T. tuberculata DL was hypothesized as a cryptic species of T. tuberculata. Meanwhile, T. tuberculata NY and T. tuberculata JN were hypothesized as potential cryptic species of T. tuberculata in this study. In both BI and ML trees, the monophyly of Scutigeromorpha, Scolopendromorpha, Geophilomorpha, and Lithobiomorpha was forcefully advocated. Moreover, Scutigeromorpha was recovered as the sister clade of (Scolopendromorpha + (Lithobiomorpha + Geophilomorpha)). Four specimens of T. tuberculata were clustered into one clade, which was the sister to the clade of S. coleoptrata. [ABSTRACT FROM AUTHOR]

The complete mitochondrial (mt) genome of Leptomantella tonkinae (Hebard, 1920) was 15,527 bp in length and contained 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and one control region. The gene arrangement of mt genome of L. tonkinae was identical to the primitive mantis. The overall AT content of the mt genome was 74%. In ML and BI phylogenetic analyses, the monophyly of Leptomantellidae was robustly supported and the clade of Leptomantellidae is a sister clade to the group of (Gonypetidae+(Leptomantellidae+(Amorphoscelidae+Nanomantidae))). [ABSTRACT FROM AUTHOR]

Rana johnsi (Smith 2009) firstly considered as the member of genus Pseudorana, has been moved into the genus Rana. In this study, we sequenced the complete mitochondrial (mt) genome of R. johnsi using the Sanger method. The circular mt genome was 17,873 bp in length and contains 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosome RNA genes, and one control region. The overall nucleotide composition in majority-strand was 28% A, 29% T, 29% C, and 14% G. We discussed the phylogenetic relationship of R. johnsi in genus Rana using ML and BI analyses based on 13 PCGs. Excluding the clade of subgenus Lithobates, Rana draytonii was the basal clade to all other Rana species, which included R. johnsi as the basal clade. The monophyly of genus Rana was supported, whereas Pseudorana was failed to support. [ABSTRACT FROM AUTHOR]

The Lonchodinae (Phasmatodea: Phasmatidae) is rich in insect species with more than 330 species of 40 genera. The phylogenetic relationships within Lonchodinae have been under debate. We successfully sequenced the complete mitogenome of Eurycantha calcarata Lucas, 1869 (Phasmatodea: Lonchodinae) with a length of 16,280 bp, which had the same genes and gene arrangements as those of various published papers on stick insects. The whole mitogenome and control region of E. calcarata had a high AT content of 78.2 and 85.9%, respectively. All PCGs used ATN as the start codon, and most PCGs used TAA/TAG as the stop codons excluding COX2 (T), COX3 (TA), and ND5 (TA). To discuss the phylogeny of Lonchodinae, we reconstructed the phylogenetic relationships of 27 species of Phasmatodea including E. calcarata and two species of Embioptera used as outgroups. In BI and ML trees, the monophyly of Lonchodinae and Necrosciinae was well supported, whereas the monophyly of Clitumninae was not recovered. These results indicated that Lonchodinae was a sister clade to Phylliinae and E. calcarata was a sister clade to Phraortes genus. [ABSTRACT FROM AUTHOR]

The first complete mitochondrial genome of Euroleon coreanus (Okamoto, 1926) was 15,797 bp in length, and contained 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and the control region. Compared to the classic insect mitochondrial genome, E. coreanus showed a gene rearrangement of ND2-C-W-Y-COX1. The overall AT content of the mitochondrial genome was 75.5%. The monophyly of Ascalaphidae, Myrmeleontidae, Nemopteridae, Nymphidae, and Psychopsidae was supported in both BI and ML trees. And E. coreanus was a sister clade to the clade of genus Myrmeleon. [ABSTRACT FROM AUTHOR]

Dryophytes versicolor is one of the most extreme freeze-tolerant frogs from eastern North America. In this study, the mitochondrial genome of D. versicolor was sequenced to analyze the phylogenetic relationships among Hylidae and investigate mitochondrial gene expression in response to freezing and anoxia. The total length of the D. versicolor mitogenome is the longest known to date among the available family members of Hylidae. Both maximum likelihood (ML) and Bayesian inference (BI) analyses strongly supported D. versicolor as a sister clade to (D. japonica + D. ussuriensis) + (D. suweonensis + D. immaculata (KP212702)), and indicated that Dryophytes is monophyletic. Using the mitochondrial genome, gene expression analysis was performed using RT-qPCR in skeletal muscle samples, and determined that relative levels of D. versicolor COX2 increased by 2.40 ± 0.23 fold in response to anoxia, but did not change with exposure to freezing. In addition, ND3 transcript levels decreased in response to anoxia but remained constant during freezing. By contrast, COX1 transcript levels decreased with exposure to freezing, but did not change under anoxic conditions. These results suggest that modulations of protein-coding mitochondrial genes of D. versicolor may play a role in the molecular response to freezing and anoxia tolerance. [ABSTRACT FROM AUTHOR]

The mitochondrial genome sequence of Xanthomantis bimaculata (Mantodea: Iridopterygidae) from Yunnan, China is a circular molecule with the typical insect mitochondrial gene arrangement, which is 15,941 bp in length and contains 22 tRNAs, two rRNAs, 13 protein-coding genes, and one control region. The overall AT content of the mitogenome is 73.12% (A = 37.58%, T = 35.54%, C = 16.54%, G = 10.34%). In BI and ML phylogenetic analyses, X. bimaculata was a sister clade to Sceptuchus simplex. The monophyly of the families Iridopterygidae, Thespidae and Liturgusidae were supported. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of Pedetontus zhejiangensis (Microcoryphia: Machilidae) was successfully sequenced. The mitochondrial genome of P. zhejiangensis was a circular molecule of 15,602 bp in length, containing 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and the control region, which showed the typical insect mitochondrial genome arrangement. The AT content of the whole genome was 73.8% and the length of the control region was 671 bp with 82.5% AT content. In BI and ML phylogenetic trees, P. zhejiangensis was a sister group to Pedetontus silvestrii, and the monophyly of Pedetontus was strongly supported. The genus Pedetontinus was a sister group to Pedetontus. [ABSTRACT FROM AUTHOR]

The mitochondrial genome of Teratoscincus roborowskii (Squamata: Gekkonidae) (GenBank No. MT107158) was a circular molecule of 16,693 bp in length, including 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and 1 non-coding region (control region). The overall base composition of the H-strand of T. roborowskii is 30.6% A, 25.6% T, 29.7% C, 14.1% G, respectively. Phylogenetic analyses showed that T. roborowskii (MT107158) was a sister clade to T. keyserlingii whereas Teratoscincus roborowskii (KP115216) was a close sister clade of T. keyserlingii and T. roborowskii (MT107158). [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of the Annamanum lunulatum is 15,610 bp in length, which contains 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and the A + T-rich region. The arrangement of genes is identical to all know longhorn beetles mitochondrial genomes. The overall AT content of the mitochondrial genome is 75.3%, whereas the AT content of A + T-rich region is 84.3%. In ML and BI phylogenetic analyses, A. lunulatum is a sister clade to Blepephaeus succinctor, and the monophyly of Lamiinae is supported. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of Mantis religiosa (Mantodea: Mantidae) from Canada was successfully sequenced. The mitochondrial genome was a circular molecule of 15,521 bp in length, containing 13 protein-coding genes, two rRNA genes, 23 tRNA genes (including an extra tRNAArg gene), and the control region. The AT content of the whole genome was 76.9% and the length of the control region was 636 bp with 81.9% AT content. The structure of the M. religiosa mitochondrial genome from Canada was almost identical to M. religiosa from China and their genetic distance was just 0.017. In Bayesian inference (BI) and maximum likelihood (ML) analyses, we found that M. religiosa was a sister clade to Statilia sp. and the monophyly of the genera Hierodula and Rhombodera was not supported. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of Chrysopa pallens (Neuroptera, Chrysopidae) was sequenced. It consists of 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA (rRNA) genes, and a control region (AT-rich region). The total length of C. pallens mitogenome is 16,723 bp with 79.5% AT content, and the length of control region is 1905 bp with 89.1% AT content. The non-coding regions of C. pallens include control region between 12S rRNA and trnI genes, and a 75-bp space region between trnI and trnQ genes. [ABSTRACT FROM AUTHOR]

Abstract Different mitochondrial gene copies or gene rearrangements are observed in some interspecific mitochondrial genomes (mitogenomes), but few in intraspecific mitogenomes. In this study, we sequenced the complete mitogenomes of Statilia maculata and Statilia nemoralis (Mantinae: Mantini). The genetic distance of the whole genomes between these two species was just 0.5%, suggesting that S. maculata and S. nemoralis might be the same species. The mitogenome of S. maculata had five copies of trnR genes (three copies were identical and the other two had some base substitutions), whereas the mitogenome of S. nemoralis had six trnR genes. Five, six or seven tandem copies of trnR genes in S. maculata were found in different localities, respectively. We further sequenced the ND3 - ND5 region of fifty individuals hatched from one ootheca (Tianjin) and we found that all individuals had five identical tandem copies of trnR genes and same sequences. Hence, we concluded that different copies of trnR genes can occur in the same species. The mitogenomes of S. maculata and S. nemoralis with other mitogenomes of Mantinae species published were used to construct BI and ML phylogenetic trees. In this study, both BI and ML phylogenetic trees showed that Mantini and Statilia were monophyletic whereas Paramantini was paraphyletic. [ABSTRACT FROM AUTHOR]