Metazoan Mitochondrial Genomes Research Articles

ORFans in mitochondrial genomes of marine polychaete Polydora.

Most characterised metazoan mitochondrial genomes are compact and encode a small set of proteins that are essential for oxidative phosphorylation, as well as rRNA and tRNA for their expression. However, in rare cases, invertebrate taxa have additional open reading frames in their mtDNA sequences. Here, we sequenced and analysed the mitochondrial genome of a polychaete worm, Polydora cf. ciliata, part of whose life cycle takes place in low-oxygen conditions. In the mitogenome, we found three "ORFan" regions (544, 1060, and 427 bp) that have no resemblance to any standard metazoan mtDNA gene but lack stop codons in one of the reading frames. Similar regions are found in the mitochondrial genomes of three other Polydora species and Bocardiella hamata. All five species share the same gene order in their mitogenomes, which differ from that of other known Spionidae mitogenomes. By analysing the ORFan sequences, we found that they are under purifying selection pressure and contain conservative regions. The codon adaptation indices (CAIs) of the ORFan genes were in the same range of values as the CAI of conventional protein-coding genes in corresponding mitochondrial genomes. The analysis of the Polydora cf. ciliata mitochondrial transcriptome showed that ORFan-544, ORFan-427 and a portion of the ORFan-1060 are transcribed. Together, this suggests that ORFan-544 and ORFan-427 encode functional proteins. It is likely that the ORFans originated when the Polydora/Bocardiella species complex separated from the rest of the Spionidae, and this event coincided with massive gene rearrangements in their mitochondrial genomes and tRNA-Met duplication.

Characterization of the complete mitochondrial genome of the striped soldier shrimp, Plesionika edwardsii (Brandt, 1851) (Crustacea: Decapoda: Pandalidae), and comparison with other species of Caridea

Abstract The striped soldier shrimp, Plesionika edwardsii (Brandt, 1851) is a pandalid with economic value in the Mediterranean region. We have sequenced and assembled its complete mitochondrial genome, which is 15,956 bp in length and contains the same 37 genes found in most metazoan mitochondrial genomes. Its gene order and nucleotide content are similar to most of the caridean mitochondrial genomes. In the comparative analysis, however, we detected in other species changes in the gene order that could be mediated by the recombination of transfer RNA genes, as well as AT skew shifts that could indicate changes in the origins of replication. All protein-coding genes of the mitochondrial genome of P. edwardsii seem to be under purifying selection, although the differences in Ka:Ks ratios suggest a disparity in the mutational constraints of some genes. This genome also presents a 1,118 bp-long non-coding sequence that encompass the control region. We have been able to find a previously described conserved sequence block in this region and assess that it forms a stem-loop structure in different species of Pandalidae, which is a shared feature with the conserved sequence blocks described in the family Alvinocarididae. We also detected microsatellites in the control region of P. edwardsii and in other species of Pandalidae and minisatellites in Lysmata vittata (Stimpson, 1860) that can account for around 20% of the additional non-coding region of this species. The phylogenetic relationships of P. edwardsii with other pandalids were assessed by two analyses: one based on the complete mitochondrial sequences and another based only on the protein-coding genes. Our study, thus, contributes to the genomic resources available for P. edwardsii and expands the current biological knowledge about the mitochondrial genomes of other caridean species.

Inverted base composition skews and discontinuous mitochondrial genome architecture evolution in the Enoplea (Nematoda)

BackgroundWithin the class Enoplea, the earliest-branching lineages in the phylum Nematoda, the relatively highly conserved ancestral mitochondrial architecture of Trichinellida is in stark contrast to the rapidly evolving architecture of Dorylaimida and Mermithida. To better understand the evolution of mitogenomic architecture in this lineage, we sequenced the mitogenome of a fish parasite Pseudocapillaria tomentosa (Trichinellida: Capillariidae) and compared it to all available enoplean mitogenomes.ResultsP. tomentosa exhibited highly reduced noncoding regions (the largest was 98 bp), and a unique base composition among the Enoplea. We attributed the latter to the inverted GC skew (0.08) in comparison to the ancestral skew in Trichinellidae (-0.43 to -0.37). Capillariidae, Trichuridae and Longidoridae (Dorylaimida) generally exhibited low negative or low positive skews (-0.1 to 0.1), whereas Mermithidae exhibited fully inverted low skews (0 to 0.05). This is indicative of inversions in the strand replication order or otherwise disrupted replication mechanism in the lineages with reduced/inverted skews. Among the Trichinellida, Trichinellidae and Trichuridae have almost perfectly conserved architecture, whereas Capillariidae exhibit multiple rearrangements of tRNA genes. In contrast, Mermithidae (Mermithida) and Longidoridae (Dorylaimida) exhibit almost no similarity to the ancestral architecture.ConclusionsLongidoridae exhibited more rearranged mitogenomic architecture than the hypervariable Mermithidae. Similar to the Chromadorea, the evolution of mitochondrial architecture in enoplean nematodes exhibits a strong discontinuity: lineages possessing a mostly conserved architecture over tens of millions of years are interspersed with lineages exhibiting architectural hypervariability. As Longidoridae also have some of the smallest metazoan mitochondrial genomes, they contradict the prediction that compact mitogenomes should be structurally stable. Lineages exhibiting inverted skews appear to represent the intermediate phase between the Trichinellidae (ancestral) and fully derived skews in Chromadorean mitogenomes (GC skews = 0.18 to 0.64). Multiple lines of evidence (CAT-GTR analysis in our study, a majority of previous mitogenomic results, and skew disruption scenarios) support the Dorylaimia split into two sister-clades: Dorylaimida + Mermithida and Trichinellida. However, skew inversions produce strong base composition biases, which can hamper phylogenetic and other evolutionary studies, so enoplean mitogenomes have to be used with utmost care in evolutionary studies.

The mitochondrial genome of Grapsus albolineatus (Decapoda: Brachyura: Grapsidae) and phylogenetic associations in Brachyura

Complete mitochondrial genomes (mitogenomes) can provide useful information for phylogenetic relationships, gene rearrangement, and evolutionary traits. In this study, we determined the complete mitochondrial DNA sequence of the herbivorous crab Grapsus albolineatus. It is a typical metazoan mitochondrial genome. The total size is 15,583 bp, contains the entire set of 37 genes, and has an AT-rich region. Then, 23 of the 37 genes were encoded by the heavy (+) strand while 14 are encoded by the light (−) strand. Compared with the pan-crustacean ground pattern, two tRNA genes (tRNA-His and tRNA-Gln) were rearranged and the tandem duplication/random loss model was used to explain the observed gene rearrangements. The phylogenetic results showed that all Grapsidae crabs clustered together as a group. Furthermore, the monophyly of each family was well supported, with the exception of Menippidae. In general, the results obtained in this study will contribute to the better understanding of gene rearrangements in Grapsidae crab mitogenomes and provide new insights into the phylogeny of Brachyura.

Comparative Mitogenomics in Hyalella (Amphipoda: Crustacea).

We present the sequencing and comparative analysis of 17 mitochondrial genomes of Nearctic and Neotropical amphipods of the genus Hyalella, most from the Andean Altiplano. The mitogenomes obtained comprised the usual 37 gene-set of the metazoan mitochondrial genome showing a gene rearrangement (a reverse transposition and a reversal) between the North and South American Hyalella mitogenomes. Hyalella mitochondrial genomes show the typical AT-richness and strong nucleotide bias among codon sites and strands of pancrustaceans. Protein-coding sequences are biased towards AT-rich codons, with a preference for leucine and serine amino acids. Numerous base changes (539) were found in tRNA stems, with 103 classified as fully compensatory, 253 hemi-compensatory and the remaining base mismatches and indels. Most compensatory Watson–Crick switches were AU -> GC linked in the same haplotype, whereas most hemi-compensatory changes resulted in wobble GU and a few AC pairs. These results suggest a pairing fitness increase in tRNAs after crossing low fitness valleys. Branch-site level models detected positive selection for several amino acid positions in up to eight mitochondrial genes, with atp6 and nad5 as the genes displaying more sites under selection.

The complete mitochondrial genome of the freshwater crab Longpotamon kenliense (Decapoda, Brachyura, Potamidae) with phylogenetic consideration

Abstract The authors herein report for the first time the complete sequence of the mitochondrial genome of Longpotamon kenliense. The results showed that the mitochondrial genome of L. kenliense is 18,499 bp in length, shares 37 genes and 1 control region with the typical metazoan mitochondrial genome, and has a strong A + T bias (74.48%), i.e., a characteristic of the metazoan mitochondrial genome. In addition, the authors used the 13 protein-coding genes from the mitochondrial genomes of L. kenliense and 67 other species of Brachyura available from the NCBI to estimate divergence times. The divergence time of L. kenliense was 14.39 Ma, which is close to the divergence times of L. xiushuiense and L. yangtsekiense, but later than that of Sinopotamon yaanense. The results indicate that the genera Sinopotamon and Longpotamon are closely related and that the genus Sinopotamon has a longer history, which may have been caused by geological events.

The complete mitochondrial genome of Stratiolaelaps scimitus (Acari: Laelapidae) and the comparison of mitochondrial gene rearrangement in the Mesostigmata

ABSTRACTAs an important predatory mite, Stratiolaelaps scimitus (Womersley) is commercially produced to control pests of edible fungi and greenhouse. In this study, the complete mitochondrial (mt) genome of S. scimitus was sequenced by Illumina HiSeq, which comparatively analysed all reported mt genome rearrangement of mesostigmatid species. The mt genome of S. scimitus was a typical circular molecule of 16,468 bp (GenBank: MK912042) and contained the complete set of 37 genes as other metazoan mitochondrial genomes. The AT-and GC-skew on the majority strand was −0.061 and 0.365, respectively, which were reversed compared to those found in most metazoan mitochondrial genomes. Like other mesostigmatid mites, many gene rearrangements occurred in the mt genome of S. scimitus. The six mesostigmatid families containing rearranged mt genomes were characterized by unique gene order, except Diplogyniidae and Parasitidae. Shared gene clusters were found in four mesostigmatid families (Laelapidae, Blattisociidae, Phytoseiidae, and Macrochelidae), indicating that they are synapomorphic for these families. These gene characters are valuable in phylogenetic analysis of relationships at different taxonomic levels.

Improved annotation of protein-coding genes boundaries in metazoan mitochondrial genomes.

With the rapid increase of sequenced metazoan mitochondrial genomes, a detailed manual annotation is becoming more and more infeasible. While it is easy to identify the approximate location of protein-coding genes within mitogenomes, the peculiar processing of mitochondrial transcripts, however, makes the determination of precise gene boundaries a surprisingly difficult problem. We have analyzed the properties of annotated start and stop codon positions in detail, and use the inferred patterns to devise a new method for predicting gene boundaries in de novo annotations. Our method benefits from empirically observed prevalances of start/stop codons and gene lengths, and considers the dependence of these features on variations of genetic codes. Albeit not being perfect, our new approach yields a drastic improvement in the accuracy of gene boundaries and upgrades the mitochondrial genome annotation server MITOS to an even more sophisticated tool for fully automatic annotation of metazoan mitochondrial genomes.

<p class="Body"><strong>The mitochondrial genome of the red tomato spider mite, <em>Tetranychus evansi</em> Baker & Pritchard (Acari: Tetranychidae) and its implications for phylogenetic analysis </strong></p>

The red tomato spider mite, Tetranychus evansi Baker & Pritchard, is a newly emerged and globally invasive pest of solanaceous plants. In this study, the complete mitochondrial genome of T. evansi was sequenced using Illumina Hiseq technology. The complete mitochondrial genome of T. evansi is a typical circular DNA with a length of 13,064 bp, which contains 37 genes arranged in consistent with the typical metazoan mitochondrial genomes. Highly truncated mitochondrial tRNAs were observed, with 18 of the 22 tRNAs appearing to lack the D- or T- arms or both. A preliminary phylogenic analysis showed that mitochondrial genome can significantly improve the reliability of phylogenetic inference relative to short sequences, as indicated by the elevated bootstrap values. This mitochondrial genome provides a valuable source for the future phylogenetic analyses and helps clear the introduction origins of the Chinese T. evansi populations.

The complete mitochondrial genome of a blowfly Calliphora nigribarbis (Vollenhoven, 1863) (Insecta: Diptera: Calliphoridae)

In the present study, the complete mitochondrial genome of a blowfly Calliphora nigribarbis has been sequenced and analyzed. The length of complete the mitochondrial genome is 16,279 bp, with 39.50% A, 13.20% C, 9.30% G, and 38.0% T nucleotide distribution. The complete mitochondrial genome consists of 13 protein-coding genes, 22 transfer RNAs, and 2 ribosomal RNAs likewise the most metazoan mitochondrial genomes. Furthermore, phylogenetic relationships of C. nigribarbis in the subfamily Calliphorinae investigated. The results suggested that C. vomitoria is the most related species to C. nigribarbis and the genus Calliphora is not monophyletic. This study provides the first complete mitochondrial genome sequence for the species.

Complete mitochondrial genome of a troglophile Cydnidae (Hemiptera)

The complete mitochondrial genome of a specimen of Cydnidae was shotgun sequenced and each partition was characterized. This genome, with 15,289 bp in length, has all of the 37 genes commonly found in metazoan mitochondrial genomes: two for ribosomal RNAs (rRNAs), 22 for transfer RNAs (tRNAs), and 13 for proteins. Protein-coding and ribosomal genes have a similar arrangement as in other insects. Phylogenetic relationship of this species with other family groups within the infraorder Pentatomomorpha was inferred using the maximum likelihood method based on the mitogenome. The phylogenetic analysis groups this specimen with the other Cydnidae species within the Pentatomoidea clade.

The complete mitochondrial genome of a freshwater mussel Nodularia douglasiae (Bivalvia: Unionidae)

The circular F-type mitochondrial genome (15,761 bp) was completely sequenced for a Korean freshwater mussel Nodularia douglasiae (synonym Unio douglasiae; Unionidae, Unionida, Bivalvia). It contains 13 PCGs, two rRNA genes, and 22 tRNA genes, as generally shown in metazoan mitochondrial genomes. Its gene order is identical to that of F-type mitochondrial genomes observed in other freshwater mussels. With nucleotide and amino acid sequences of the complete F-type mitochondrial genomes obtained from 38 unionid species reported so far, phylogenetic analyses were done and discussed. The present study may give valuable helps to explore genetic diversity and population structures of other freshwater mussels as well as N. douglasiae.

Probing recalcitrant problems in polyclad evolution and systematics with novel mitochondrial genome resources

For their apparent morphological simplicity, the Platyhelminthes or “flatworms” are a diverse clade found in a broad range of habitats. Their body plans have however made them difficult to robustly classify. Molecular evidence is only beginning to uncover the true evolutionary history of this clade. Here we present nine novel mitochondrial genomes from the still undersampled orders Polycladida and Rhabdocoela, assembled from short Illumina reads. In particular we present for the first time in the literature the mitochondrial sequence of a Rhabdocoel, Bothromesostoma personatum (Typhloplanidae, Mesostominae). The novel mitochondrial genomes examined generally contained the 36 genes expected in the Platyhelminthes, with all possessing 12 of the 13 protein-coding genes normally found in metazoan mitochondrial genomes (ATP8 being absent from all Platyhelminth mtDNA sequenced to date), along with two ribosomal RNA genes. The majority presented possess 22 transfer RNA genes, and a single tRNA gene was absent from two of the nine assembled genomes. By comparison of mitochondrial gene order and phylogenetic analysis of the protein coding and ribosomal RNA genes contained within these sequences with those of previously sequenced species we are able to gain a firm molecular phylogeny for the inter-relationships within this clade.Our phylogenetic reconstructions, using both nucleotide and amino acid sequences under several models and both Bayesian and Maximum Likelihood methods, strongly support the monophyly of Polycladida, and the monophyly of Acotylea and Cotylea within that clade. They also allow us to speculate on the early emergence of Macrostomida, the monophyly of a “Turbellarian-like” clade, the placement of Rhabditophora, and that of Platyhelminthes relative to the Lophotrochozoa (=Spiralia). The data presented here therefore represent a significant advance in our understanding of platyhelminth phylogeny, and will form the basis of a range of future research in the still-disputed classifications within this taxon.

The complete mitochondrial genome of Somanniathelphusa boyangensis and phylogenetic analysis of Genus Somanniathelphusa (Crustacea: Decapoda: Parathelphusidae).

In this study, the authors first obtained the mitochondrial genome of Somanniathelphusa boyangensis. The results showed that the mitochondrial genome is 17,032bp in length, included 13 protein-coding genes, 2 rRNAs genes, 22 tRNAs genes and 1 putative control region, and it has the characteristics of the metazoan mitochondrial genome A+T bias. All tRNA genes display the typical clover-leaf secondary structure except tRNASer(AGN), which has lost the dihydroxyuridine arm. The GenBank database contains the mitochondrial genomes of representatives of approximately 22 families of Brachyura, comprising 56 species, including 4 species of freshwater crab. The authors established the phylogenetic relationships using the maximum likelihood and Bayesian inference methods. The phylogenetic relationship indicated that the molecular taxonomy of S. boyangensis is consistent with current morphological classification, and Parathelphusidae and Potamidae are derived within the freshwater clade or as part of it. In addition, the authors used the COX1 sequence of Somanniathelphusa in GenBank and the COX1 sequence of S. boyangensis to estimated the divergence time of this genus. The result displayed that the divergence time of Somanniathelphusa qiongshanensis is consistent with the separation of Hainan Island from mainland China in the Beibu Gulf, and the divergence time for Somanniathelphusa taiwanensis and Somanniathelphusa amoyensis is consistent with the separation of Taiwan Province from Mainland China at Fujian Province. These data indicate that geologic events influenced speciation of the genus Somanniathelphusa.

Polymorphic duplicate genes and persistent non-coding sequences reveal heterogeneous patterns of mitochondrial DNA loss in salamanders

BackgroundMitochondria are the site of the citric acid cycle and oxidative phosphorylation (OXPHOS). In metazoans, the mitochondrial genome is a small, circular molecule averaging 16.5 kb in length. Despite evolutionarily conserved gene content, metazoan mitochondrial genomes show a diversity of gene orders most commonly explained by the duplication-random loss (DRL) model. In the DRL model, (1) a sequence of genes is duplicated in tandem, (2) one paralog sustains a loss-of-function mutation, resulting in selection to retain the other copy, and (3) the non-functional paralog is eventually deleted from the genome. Despite its apparent role in generating mitochondrial gene order diversity, little is known about the tempo and mode of random gene loss after duplication events. Here, we determine mitochondrial gene order across the salamander genus Aneides, which was previously shown to include at least two DRL-mediated rearrangement events. We then analyze these gene orders in a phylogenetic context to reveal patterns of DNA loss after mitochondrial gene duplication.ResultsWe identified two separate duplication events that resulted in mitochondrial gene rearrangements in Aneides; one occurred at the base of the clade tens of millions of years ago, while the other occurred much more recently (i.e. within a single species), resulting in gene order polymorphism and paralogs that are readily identifiable. We demonstrate that near-complete removal of duplicate rRNA genes has occurred since the recent duplication event, whereas duplicate protein-coding genes persist as pseudogenes and duplicate tRNAs persist as functionally intact paralogs. In addition, we show that non-coding DNA duplicated at the base of the clade has persisted across species for tens of millions of years.ConclusionsThe evolutionary history of the mitochondrial genome, from its inception as a bacterial endosymbiont, includes massive genomic reduction. Consistent with this overall trend, selection for efficiency of mitochondrial replication and transcription has been hypothesized to favor elimination of extra sequence. Our results, however, suggest that there may be no strong disadvantage to extraneous sequences in salamander mitochondrial genomes, although duplicate rRNA genes may be deleterious.

CoreTracker: accurate codon reassignment prediction, applied to mitochondrial genomes.

Codon reassignments have been reported across all domains of life. With the increasing number of sequenced genomes, the development of systematic approaches for genetic code detection is essential for accurate downstream analyses. Three automated prediction tools exist so far: FACIL, GenDecoder and Bagheera; the last two respectively restricted to metazoan mitochondrial genomes and CUG reassignments in yeast nuclear genomes. These tools can only analyze a single genome at a time and are often not followed by a validation procedure, resulting in a high rate of false positives. We present CoreTracker, a new algorithm for the inference of sense-to-sense codon reassignments. CoreTracker identifies potential codon reassignments in a set of related genomes, then uses statistical evaluations and a random forest classifier to predict those that are the most likely to be correct. Predicted reassignments are then validated through a phylogeny-aware step that evaluates the impact of the new genetic code on the protein alignment. Handling simultaneously a set of genomes in a phylogenetic framework, allows tracing back the evolution of each reassignment, which provides information on its underlying mechanism. Applied to metazoan and yeast genomes, CoreTracker significantly outperforms existing methods on both precision and sensitivity. CoreTracker is written in Python and available at https://github.com/UdeM-LBIT/CoreTracker. mabrouk@iro.umontreal.ca. Supplementary data are available at Bioinformatics online.

The mitochondrial genomes of the acoelomorph worms Paratomella rubra, Isodiametra pulchra and Archaphanostoma ylvae

Acoels are small, ubiquitous - but understudied - marine worms with a very simple body plan. Their internal phylogeny is still not fully resolved, and the position of their proposed phylum Xenacoelomorpha remains debated. Here we describe mitochondrial genome sequences from the acoels Paratomella rubra and Isodiametra pulchra, and the complete mitochondrial genome of the acoel Archaphanostoma ylvae. The P. rubra and A. ylvae sequences are typical for metazoans in size and gene content. The larger I. pulchra mitochondrial genome contains both ribosomal genes, 21 tRNAs, but only 11 protein-coding genes. We find evidence suggesting a duplicated sequence in the I. pulchra mitochondrial genome. The P. rubra, I. pulchra and A. ylvae mitochondria have a unique genome organisation in comparison to other metazoan mitochondrial genomes. We found a large degree of protein-coding gene and tRNA overlap with little non-coding sequence in the compact P. rubra genome. Conversely, the A. ylvae and I. pulchra genomes have many long non-coding sequences between genes, likely driving genome size expansion in the latter. Phylogenetic trees inferred from mitochondrial genes retrieve Xenacoelomorpha as an early branching taxon in the deuterostomes. Sequence divergence analysis between P. rubra sampled in England and Spain indicates cryptic diversity.

Combinatorics of Tandem Duplication Random Loss Mutations on Circular Genomes

The tandem duplication random loss operation (TDRL) is an important genome rearrangement operation in metazoan mitochondrial genomes. A TDRL consists of a duplication of a contiguous set of genes in tandem followed by a random loss of one copy of each duplicated gene. This paper presents an analysis of the combinatorics of TDRLs on circular genomes, e.g., the mitochondrial genome. In particular, results on TDRLs for circular genomes and their linear representatives are established. Moreover, the distance between gene orders with respect to linear TDRLs and circular TDRLs is studied. An analysis of the available animal mitochondrial gene orders shows the practical relevance of the theoretical results.

Mitochondrial poly(A) polymerase is involved in tRNA repair.

Transcription of the mitochondrial genome results in polycistronic precursors, which are processed mainly by the release of tRNAs interspersed between rRNAs and mRNAs. In many metazoan mitochondrial genomes some tRNA genes overlap with downstream genes; in the case of human mitochondria the genes for tRNATyr and tRNACys overlap by one nucleotide. It has previously been shown that processing of the common precursor releases an incomplete tRNATyr lacking the 3′-adenosine. The 3′-terminal adenosine has to be added before addition of the CCA end and subsequent aminoacylation. We show that the mitochondrial poly(A) polymerase (mtPAP) is responsible for this A addition. In vitro, a tRNATyr lacking the discriminator is a substrate for mtPAP. In vivo, an altered mtPAP protein level affected tRNATyr maturation, as shown by sequencing the 3′ ends of mitochondrial tRNAs. Complete repair could be reconstituted in vitro with three enzymes: mtPAP frequently added more than one A to the 3′ end of the truncated tRNA, and either the mitochondrial deadenylase PDE12 or the endonuclease RNase Z trimmed the oligo(A) tail to a single A before CCA addition. An enzyme machinery that evolved primarily for other purposes thus allows to tolerate the frequent evolutionary occurrence of gene overlaps.

Characterization of the complete mitochondrial genomes from Polycladida (Platyhelminthes) using next-generation sequencing

The complete mitochondrial genomes of three polycladids, the acotylean Hoploplana elisabelloi and the cotyleans Enchiridium sp. and Prosthiostomum siphunculus have been assembled with high coverage from Illumina sequencing data. The mt genomes contain 36 genes including 12 of the 13 protein-coding genes characteristic for metazoan mitochondrial genomes, two ribosomal RNA genes, and 22 transfer RNA genes. Gene annotation, gene order, genetic code, start and stop codons and codon bias have been identified. In comparison with the well investigated parasitic Neodermata, our analysis reveals a great diversity of gene orders within Polycladida and Platyhelminthes in general. By analyzing representative genomes of the main groups of Platyhelminthes we explored the phylogenetic relationships of this group. The phylogenetic analyses strongly supported the monophyly of Polycladida, and based on a small taxon sampling suggest the monophyly of Acotylea and Cotylea.