Comprehensive Draft Genome Analyses of Three Rockfishes (Scorpaeniformes,Sebastiscus) via Genome Survey Sequencing

Sebastiscus species, marine rockfishes, are of essential economic value. However, the genomic data of this genus is lacking and incomplete. Here, whole genome sequencing of all species of Sebastiscus was conducted to provide fundamental genomic information. The genome sizes were estimated to be 802.49 Mb (S. albofasciatus), 786.79 Mb (S. tertius), and 776.00 Mb (S. marmoratus) by using k-mer analyses. The draft genome sequences were initially assembled, and genome-wide microsatellite motifs were identified. The heterozygosity, repeat ratios, and numbers of microsatellite motifs all suggested possibly that S. tertius is more closely related to S. albofasciatus than S. marmoratus at the genetic level. Moreover, the complete mitochondrial genome sequences were assembled from the whole genome data and the phylogenetic analyses genetically supported the validation of Sebastiscus species. This study provides an important genome resource for further studies of Sebastiscus species.     

Genetic and morphological identification of Sebastiscus tertius in the East China Sea (Scorpaeniformes: Scorpaenidae)

The allozymes and morphology of 110 specimens of three Sebastiscus species (S. marmoratus, S. tertius, and S. albofasciatus) in the East China Sea and near Japan were compared. Results of 20 allozyme loci studied showed that all three species were closely related (Nei's unbiased genetic distances, 0.057–0.133) but could be identified on the basis of informative loci with a few exceptions. Initial identification based on color patterns agreed with allozyme identification in more than 98% of Sebastiscus specimens and agreed completely in S. albofasciatus. One specimen that was initially identified as S. marmoratus because of the dark body color was actually S. tertius according to two informative allozyme loci. Number of pectoral fin rays differed between S. marmoratus (18 or fewer, 98%) and S. tertius (19 or more, 85%) in this study. The previously mentioned dark specimen had 19 pectoral fin rays, which are characteristic in S. tertius. Using seven morphological measurements, canonical discriminant analysis between S. marmoratus and S. tertius classified less than 90% of specimens into the original groups (species). Some specimens of S. tertius resembled S. marmoratus in body shape and vice versa. A combination of genetic characterization and morphological examination is necessary to identify S. marmoratus and S. tertius accurately. Distinction based on allozymes and color patterns with numbers of pectoral fin rays should provide satisfactory identification. Received: May 16, 2000 / Revised: December 19, 2000 / Accepted: January 24, 2001     

Mitochondrial genomes of five Hyphessobrycon tetras and their phylogenetic implications

To date, the taxonomic status and phylogenetic affinities within Hyphessobrycon, even among other genera in Characidae, remain unclear. Here, we determined five new mitochondrial genomes (mitogenomes) of Hyphessobrycon species (H. elachys, H. flammeus, H. pulchripinnis, H. roseus, and H. sweglesi). The mitogenomes were all classical circular structures, with lengths ranging from 16,008 to 17,224 bp. The type of constitutive genes and direction of the coding strand that appeared in the mitogenomes were identical to those of other species in Characidae. The highest value of the Ka/Ks ratio within 13 protein‐coding genes (PCGs) was found in ND2 with 0.83, suggesting that they were subject to purifying selection in the Hyphessobrycon genus. Comparison of the control region sequences among seven Hyphessobrycon fish revealed that repeat units differ in length and copy number across different species, which led to sharp differences in mitogenome sizes. Phylogenetic trees based on the 13 PCGs did not support taxonomic relationships, as the Hyphessobrycon fish mixed with those from other genera. These data were combined to explore higher level relationships within Characidae and could aid in the understanding of the evolution of this group.     

Sequence comparison of the mitochondrial genomes of five brackish water species of the family Neritidae: Phylogenetic implications and divergence time estimation

Neritids are ancient gastropod species which can live in marine, brackish water, and freshwater environments. In this study, we sequenced and annotated the mitochondrial genomes of five brackish water neritids (i.e., Clithon corona, Clithon lentiginosum, Clithon squarrosum, Neritina iris, and Septaria lineata). The mitogenomes ranged from 15,618 to 15,975 bp, and all contain 13 protein‐coding genes (PCGs), 22 tRNA genes, and two rRNA genes, with a closed ring structure. We calculated the Ka/Ks values of all 13 PCGs of Neritidae species, all ratios are less than 1, under purification selection. Phylogenetic analysis of the 13 PCGs showed that Neritimorpha is a sister group with Vetigastropoda and Caenogastopoda, genus Clithon is a sister group with Neritina and Septaria. Estimation of divergence time for all species of Neritidae showed that the main differentiation of Neritidae occurred in Cenozoic period (65 Mya), C. corona and C. lentiginosum were differentiated in the Cenozoic Neogene, the other three species diverged in the Cenozoic Paleogene. These results will help to better understand the evolutionary position of Neritidae and provide reference for further phylogenetic research on Neritidae species.     

The first complete mitogenome of Picumnus innominatus (Aves,Piciformes, Picidae) and phylogenetic inference within the Picidae

The avian family Picidae, which is nearly global in distribution, contains the piculets (Picumninae and Nesoctitinae), the woodpeckers (Picinae), and the wrynecks (Jynginae). However, the phylogenetic relationships within the Picidae remain obscure for most genera. In the present study, the complete mitochondrial genome of Picumnus innominatus was determined and described, which was the first complete mitogenome reported in the Picumnus. The circular mitogenome of P. innominatus was 17,180 bp in size and consisted of 13 protein-coding genes (PCGs), 22 tRNAs, 2 rRNA genes, a control region (CR), and a noncoding region. The gene order and arrangement of the P. innominatus mitogenome were identical to other mitogenomes of the Picidae. Moreover, strikingly large tandem repeats were found in the noncoding region of the P. innominatus mitogenome, which have not yet been covered in other picid species to date. At the family level (Picidae), the highest dN/dS ratio was detected for the ND1 gene (1.38726) among 13 PCGs, indicating that positive selection was powerful for this gene. Bayesian and Maximum Likelihood phylogenetic analyses based on the combination of 12S rRNA and CYTB gene supported strongly that the Picumninae is monophyletic.     

The complete mitochondrial genome of <Emphasis Type="Italic">Vanessa indica</Emphasis> and phylogenetic analyses of the family Nymphalidae

Vanessa indica is a small butterfly lacking historical molecular and biological research. Vanessa indica belongs to the family Nymphalidae (Lepidoptera: Papilionoidea), which is the largest group of butterflies and are nearly ubiquitous. However, after more than a century of taxonomic and molecular studies, there is no consensus for family classification, and the phylogenetic relationships within Nymphalidae are controversial. The first objective was to sequence and characterize the complete mitochondrial genome of V. indica. The most important objective was to completely reconstruct the phylogenetic relationships for family members within Nymphalidae. The mitochondrial genomic DNA (mtDNA) of V. indica was extracted and amplified by polymerase chain reaction. The complete mitochondrial sequence was annotated and characterized by analyzing sequences with SeqMan program. The phylogenetic analyses were conducted on thirteen protein coding genes (PCGs) in 95 mtDNA of Nymphalidae downloaded from GenBank for reference using the maximum likelihood method and Bayesian inference to ensure the validity of the results. The complete mitogenome was a circular molecule with 15,191 bp consisting of 13 protein coding genes, two ribosomal RNA genes (16S rRNA and 12S rRNA), 22 transfer RNA (tRNA) genes, and an A?+?T-rich region (D-loop). The nucleotide composition of the genome was highly biased for A?+?T content, which accounts for 80.0% of the nucleotides. All the tRNAs have putative secondary structures that are characteristic of mitochondrial tRNAs, except tRNA^Ser(AGN). All the PCGs started with ATN codons, except cytochrome c oxidase subunit 1 (COX1), which was found to start with an unusual CGA codon. Four genes were observed to have unusual codons: COX1 terminated with atypical TT and the other three genes terminated with a single T. The A?+?T rich region of 327 bp consisted of repetitive sequences, including a ATAGA motif, a 19-bp poly-T stretch, and two microsatellite-like regions (TA)₈. The phylogenetic analyses consistently placed Biblidinae as a sister cluster to Heliconiinae and Calinaginae as a sister clade to Satyrinae. Moreover, the phylogenetic tree identified Libytheinae as a monophyletic group within Nymphalidae. The complete mitogenome of V. indica was 15,191 bp with mitochondrial characterizations common for lepidopteran species, which enriched the mitochondria data of Nymphalid species. And the phylogenetic analysis revealed different classifications and relationships than those previously described. Our results are significant because they would be useful in further understanding of the evolutionary biology of Nymphalidae.     

Comparative mitogenome analysis of three species and monophyletic inference of Catantopinae (Orthoptera: Acridoidea)

To enrich the genomic database of Catantopinae (Orthoptera: Acrididae), mitogenomes of three species from different genera, Traulia nigritibialis (15,701 bp), Choroedocus capensis (16,293 bp) and Stenocatantops splendens (15,574 bp), were characterized and compared with those of other grasshoppers in the subfamily. All 13 protein-coding genes (PCGs) were initiated by ATN codons except COI with ACC (C. capensis and S.splendens) and ND6 with TTG (S. splendens). All transfer RNA (tRNA) genes had a typical clover-leaf structure, except tRNA^Ser(AGN) in which the base pairs of the dihydrouridine (DHU) arm were reduced. The phylogenetic relationships were constructed among 22 species from four subfamiles of Acrididae by classical classifications based on two datasets of their mitogenomes using both Bayesian Inference (BI) and Maximum Likelihood (ML). The phylogenetic analysis confirmed the monophyly of the three other subfamilies, but did not provided support of the monophyly of Catantopinae.     

Phylogenetic relationships among four species of Mullidae (Perciformes) inferred from DNA sequences of mitochondrial cytochrome b and 16S rRNA genes

DNA sequence comparisons of two mitochondrial DNA genes were used to infer phylogenetic relationships among four species of mullids. Approximately 238 bp of the mitochondrial 16S ribosomal RNA (rRNA) and 261 bp of the cytochrome b (cytb) genes were sequenced from representatives of three mullid genera (Mullus, Upeneus, Pseudopeneus), present in the Mediterranean Sea. Trees were constructed using three methods: maximum likelihood (ML), neighbor joining (NJ) and parsimony (MP). The results of the analyses of these data together with published data of the same mtDNA segments of two other perciform species (Sparus aurata, Perca fluviatilis), support the previous taxonomic classification of the three genera examined, as well as the classification of the two red mullet species in the same genus.     

The complete mitochondrial genome sequence of the little egret (Egretta garzetta)

Many phylogenetic questions in the Ciconiiformes remain unresolved and complete mitogenome data are urgently needed for further molecular investigation. In this work, we determined the complete mitogenome sequence of the little egret (Egretta garzetta). The genome was 17,361 bp in length and the gene organization was typical of other avian mtDNA. In protein-coding genes (PCGs), a C insertion was found in ND3, and COIII and ND4 terminated with incomplete stop codons (T). tRNA-Val and tRNA-Ser (AGY) were unable to fold into canonical cloverleaf secondary structures because they had lost the DHU arms. Long repetitive sequences consisting of five types of tandem repeats were found at the 3′ end of Domain III in the control region. A phylogenetic analysis of 11 species of Ciconiiformes was done using complete mitogenome data and 12 PCGs. The tree topologies obtained with these two strategies were identical, which strongly confirmed the monophyly of Ardeidae, Threskiorothidae and Ciconiidae. The phylogenetic analysis also revealed that Egretta was more closely related to Ardea than to Nycticorax in the Ardeidae, and Platalea was more closely related to Threskiornis than to Nipponia in the Threskiornithidae. These findings contribute to our understanding of the phylogenetic relationships of Ciconiiformes based on complete mitogenome data.     

Mitochondrial genome characteristics of Somena scintillans (Lepidoptera: Erebidae) and comparation with other Noctuoidea insects

In this study, mitogenome of Somena scintillans (Lepidoptera: Erebidae) were sequenced and compared with other Noctuoidea species. The mitogenome is 15,410 base pairs in length. All 13 protein-coding genes (PCGs) are initiated by ATN codons except cox1 with CGA and all of PCGs terminate with TAA except nad4 with TAG. The codons ACG and CGC are absent. All the tRNA genes could be folded into the typical cloverleaf secondary structure except the trnS1 which not only loses dihydrouridine (DHU) arm but also mutates its anticodon into TCT. In the AT-rich region of the mitogenome the motif ‘ATAGA’ mutates to ‘ATATA’ and two copies of 161 bp-tandem repeats and two ‘TA’ short tandem repeats are founded. Phylogenetic analyses showed that S. scintillans is clustered into subfamily Lymatriinae. The phylogenetic relationships within Noctuoidea is (((Nolidae + (Euteliidae + Noctuidae)) + Erebidae) + Notodontidae)     

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.     

Delimiting priorities while biodiversity is lost: Rio’s seasonal killifishes on the edge of survival

The coastal plains of the Rio de Janeiro State, south-eastern Brazil, have been considered an important hotspot of aplocheiloid killifish diversity. A conservation status assessment based on 17?years of field studies directed to record habitat decline indicates that five species of seasonal killifish, Leptolebias marmoratus, Nematolebias papilliferus, Notholebias cruzi, Notholebias fractifasciatus and Ophthalmolebias constanciae, distributed among three areas of endemism, are on the edge of survival. Conservation priority, focusing on five remnant isolated locations where those species occur, was evaluated on the basis of the phylogenetic diversity content of each location established through a phylogenetic base containing data on relationships of 118 cynolebiasine species, including all species endemic to Rio. Data on species diversity for each area and data on morphological uniqueness were subsequently compared and discussed. The Barra de S?o Jo?o pool, in the S?o Jo?o-Cabo Frio area, habitat of Nematolebias whitei, No. cruzi, O. constanciae, concentrates the highest value for conservation proposal, followed by the Ino? pool, in the Maricá area, habitat of Ne. papilliferus and No. fractifasciatus. However, in case of the Barra de S?o Jo?o pool being effectively preserved, it is recommended to concentrate efforts to preserve the Cava swamp as a second priority, since although the latter location showing the lowest individual value, Cava swamp and Barra de S?o Jo?o pool together sum a higher phylogenetic diversity index by preserving more distinct cynolebiasine lineages.     

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.     

Phylogenetic estimation of Mytilidae in the East China Sea inferred from mitochondrial genes and nuclear DNA sequence variation

We performed a phylogenetic estimation of the family Mytilidae in the East China Sea based on nuclear internal transcribed spacer (ITS) genes and two mitochondrial genes (COI and 16S RNA). Analysis of five mytilid species based on each of the three genes resulted in mostly congruent trees, although there were some discrepancies in the classification of these species. We combine the results obtained from the three separate analyses to provide a phylogenetic estimation of Mytilidae. We found that the Mytilidae was divided into two major lineages: in one clade, Mytilus galloprovincialis was grouped with Mytilus coruscus; in the second clade, Septifer bilocularis was placed at the basal position in an individual clade, and Perna viridis and Musculista senhousia were recovered as a monophyletic group. Although these finding provide important insights into the taxonomic relationships among the Mytilidae, many aspects of Mytilidae phylogeny remain unresolved. Further analysis based on more molecular information and extensive taxon sampling is necessary to elucidate the phylogenetic relationships among the major lineages within the Mytilidae.     

Characteristics of the mitochondrial genome of Rana omeimontis and related species in Ranidae: Gene rearrangements and phylogenetic relationships

The Omei wood frog (Rana omeimontis), endemic to central China, belongs to the family Ranidae. In this study, we achieved detail knowledge about the mitogenome of the species. The length of the genome is 20,120 bp, including 13 protein‐coding genes (PCGs), 22 tRNA genes, two rRNA genes, and a noncoding control region. Similar to other amphibians, we found that only nine genes (ND6 and eight tRNA genes) are encoded on the light strand (L) and other genes on the heavy strand (H). Totally, The base composition of the mitochondrial genome included 27.29% A, 28.85% T, 28.87% C, and 15.00% G, respectively. The control regions among the Rana species were found to exhibit rich genetic variability and A + T content. R. omeimontis was clustered together with R. chaochiaoensis in phylogenetic tree. Compared to R. amurensis and R. kunyuensi, it was more closely related to R. chaochiaoensis, and a new way of gene rearrangement (ND6‐trnE‐Cytb‐D‐loop‐trnL2 (CUN)‐ND5‐D‐loop) was also found in the mitogenome of R. amurensis and R. kunyuensi. Our results about the mitochondrial genome of R. omeimontis will contribute to the future studies on phylogenetic relationship and the taxonomic status of Rana and related Ranidae species.     

The first complete mitochondrial genome of the Indian Tent Turtle,Pangshura tentoria (Testudines: Geoemydidae): Characterization and comparative analysis

The characterization of a complete mitogenome is widely used in genomics studies for systematics and evolutionary research. However, the sequences and structural motifs contained within the mitogenome of Testudines taxa have rarely been examined. The present study decodes the first complete mitochondrial genome of the Indian Tent Turtle, Pangshura tentoria (16,657 bp) by using next‐generation sequencing. This denovo assembly encodes 37 genes: 13 protein‐coding genes (PCGs), 22 transfer RNA (tRNAs), two ribosomal RNA, and one control region (CR). Most of the genes were encoded on majority strand, except for one PCG (NADH dehydrogenase subunit 6) and eight tRNAs. Most of the PCGs were started with an ATG initiation codon, except for Cytochrome oxidase subunit 1 with “GTG” and NADH dehydrogenase subunit 5 with “ATA.” The termination codons, “TAA” and “AGA” were observed in two subunits of NADH dehydrogenase gene. The relative synonymous codon usage analysis revealed the maximum abundance of alanine, isoleucine, leucine, and threonine. The nonsynonymous/synonymous ratios were <1 in all PCGs, which indicates strong negative selection among all Geoemydid species. The study also found the typical cloverleaf secondary structure in most of the tRNA genes, except for serine with the lack of the conventional DHU arm. The comparative study of Geoemydid mitogenomes revealed the occurrence of tandem repeats was frequent in the 3′ end of CR. Further, two copies of a unique tandem repeat “TTCTCTTT” were identified in P. tentoria. The Bayesian and maximum‐likelihood phylogenetic trees using concatenation of 13 PCGs revealed the close relationships of P. tentoria with Batagur trivittata in the studied dataset. All the Geoemydid species showed distinct clustering with high bootstrap support congruent with previous evolutionary hypotheses. We suggest that the generations of more mitogenomes of Geoemydid species are required, to improve our understanding of their in‐depth phylogenetic and evolutionary relationships.     

Genetic diversity, population structure and phylogenetic inference among Italian Orchids of the Serapias genus assessed by AFLP molecular markers

The means by which new species arise remain a fundamental question in the Orchid family evolution. Even if the Orchid-pollinator interactions are highly specific, events of cross-hybridization in Serapias are possible and represent a central issue of the plant's evolutionary biology. Italian Orchids have a wide variety of pollination systems and highly diverse floral traits, considered as some of the main tools for activating the speciation processes. The assess of diversity in Serapias taxa and eventual phylogenetic relationships among Serapias lingua, S. parviflora, S. vomeracea, S. apulica, S. nurrica, S. neglecta, S. cordigera, S. politisii, and some putative inter-specific hybrids were investigated at the DNA level by AFLP markers. The present molecular study on Serapias populations (1) confirms some previous finding, (2) opens new hypotheses on the possible evolutionary and taxonomic relationships existing among the Serapias species, and (3) gives indications about the real consistence of hybrid individuals in a genus characterized by a relevant gene flow.     

The complete mitochondrial genome of Tonkinacris sinensis(Orthoptera: Acrididae): A tRNA-like sequence and its implications for phylogeny

The complete mitochondrial genome of Tonkinacris sinensis is 15,627 bp long and contains13 protein-coding genes (PCGs), 22 tRNA genes, 2 rRNA genes and one A + T-rich region. The gene order and orientation are identical to those of other Orthoptera species, containing the rearrangement of trnD and trnK. Intriguingly, a tRNA^Ser-like gene exists on the N strand between the trnSUCN and nad1 genes. The length of this gene is 110 bp, and it has a typical clover-leaf structure, an anticodon, and a high cove score (23.49). On its clover-leaf structure, on the anticodon arm, there is a 41 bp intron with an unknown function. Here, phylogenetic analysis was conducted based on 13 PCGs of 30 species from 9 subfamilies of Acrididae to understand their phylogenetic relationships. According to the phylogenetic tree, the relationship among the 9 subfamilies within Acrididae was as follows: (Spathosterninae + (Oxyinae + (Catantopinae + (Calliptaminae + (Cyrtacanthacridinae + (Melanoplinae + (Gomphocerinae + (Oedipodinae + Acridinae)))))))).     

Whole mitochondrial genome of long-clawed mole vole (Prometheomys schaposchnikowi) from Turkey,with its phylogenetic relationships

The mitogenome of Prometheomys schaposchnikowi was characterized for the first time as a circular DNA molecule (16.284 bp), containing 37 coding and 2 non-coding regions. In the mitogenome, ND6 and 8 tRNA genes were encoded on the light chain, while 12 PCGs, 14 tRNAs, 2 rRNAs, D-loop and O_L were encoded on the heavy chain. The most common initiation codon in PCGs was ATG. As in many mammals, incomplete stop codons in P. schaposchnikowi were in the COX3, ND1 and ND4. Phylogenetic relationships were revealed using Bayesian method and the 13 PCGs. Seven genera (Arvicola, Dicrostonyx, Lasiopodomys, Myodes, Ondatra, Proedromys and Prometheomys) formed a monophyletic group, while Eothenomys, Microtus and Neodon were paraphyletic. P. schaposchnikowi constituted the most basal group within Arvicolinae. Divergence time estimation suggested that P. schaposchnikowi diversified during the Miocene (16.28 Mya). Further molecular studies are needed to test the distinctiveness and diversity of the genus Prometheomys.