Beata Śmietanka

Comparative Genomics of Marine Mussels (Mytilus spp.) Gender Associated mtDNA: Rapidly Evolving atp8

The unusual mode of mitochondrial DNA inheritance, with two separate: maternal (F) and paternal (M) lineages, gives unique opportunities to study the evolution of the mitochondrial genome. This system was first discovered in the marine mussels Mytilus. The three related species: Mytilus edulis, Mytilus galloprovincialis and Mytilus trossulus form a complex in which the divergence of M and F lineages pre-dates the speciation. The complete mitochondrial genomes of both lineages were known for all species except Pacific M. trossulus. Here we report, for the first time, the complete sequences of both mitochondrial genomes of Pacific M. trossulus, filling the gap. While the reported M and F genomes are highly diverged (26%), they have similar organisation. The only difference is the translocation of one tRNA gene into the long, mosaic control region of the F genome. Consistent presence of an ORF which most likely represents the atp8 gene was confirmed in both genomes. The predicted protein has characteristics expected of the functional atp8 even though the M and F versions are markedly different in length. Comparative analysis involving all three species led to the conclusion that the cause of a faster evolution of atp8 and Mytilus mtDNA in general is most likely the Compensation-Draft Feedback process coupled with relatively relaxed selection in the M lineage. Thus, we postulate that the adaptive changes may have played a role in the emergence of highly diverged, barely recognizable atp8 in Mytilus mussels.

Recombination in Mitochondrial DNA of European Mussels Mytilus

Mitochondrial DNA was long believed to be purely clonal and free from recombination. Major phylogenetic studies still depend on such assumptions. The peculiar genetic system of marine mussels Mytilus in which two divergent mitochondrial genomes exist provides a unique opportunity to study mtDNA recombination. Previous reports showed the existence of a few haplotypes having very strong recombination signal in the control region of mtDNA. Those recombinant variants have been found in a Baltic Sea population of Mytilus trossulus as well as in Mytilus galloprovincialis from the Black Sea. In both cases the mosaic genomes switched their transmission route and have been inherited paternally. In the present study rearranged mtDNA genomes found in all three European Mytilus species are described. The structure of their control region is a result of intra- and intermolecular recombination between mitochondrial genomes. Together with the phylogenetic reconstruction and geographic distribution, this suggests that two interlineage recombination events have occurred in the control region of mtDNA of European mussels Mytilus. Contrary to earlier observations, some of the mosaic genomes do not show any gender bias, which has important implications regarding the transmission and evolution of blue mussel mitochondrial genomes.

Is Interlineage Recombination Responsible for Low Divergence of Mitochondrial nad3 Genes in Mytilus galloprovincialis

The existence of mtDNA recombination in animals has been confirmed by several case studies. Still, for Mytilus mussels possessing two divergent mitochondrial genomes (M and F), which can recombine, no recombination between coding sequences of highly diverged M and F genomes has been shown. Based on the full sequences of both genomes, it has been suggested that particularly low divergence observed within the mitochondrial nad3 gene of the Mytilus galloprovincialis mussel may be caused by its exceptionally low evolutionary rate. Here, we contribute a new pair of mitochondrial genomes typical for M. galloprovincialis and show that this low divergence is not a sign of evolutionary conservation but is rather caused by the acquisition of an F-related sequence by the published M genome of M. galloprovincialis. The most likely scenario for this apparent mtDNA-coding region recombination case is an assembly artifact.

Next-generation sequencing of Dreissena polymorpha transcriptome sheds light on its mitochondrial DNA

Abstract Zebra mussels Dreissena polymorpha (Veneroida, Dreissenidae) are known for their invasive behavior. Despite numerous studies dealing with this species, no results of large sequencing projects have been published to date, hampering marker development. In this study, we present a relatively large novel transcriptomic dataset obtained by Illumina MiSeq technology from mantle and male gonad of D. polymorpha sampled in Poland. The transcriptomic data were typical for the tissue analyzed. Moreover, they showed the expression of a single mitochondrial genome, indicating that this species do not have doubly uniparental inheritance of mitochondria. The sequences of mitochondrial transcripts were used to design primers and obtain nearly complete sequence of the zebra mussel mitochondrial genome by PCR and Sanger sequencing. This mitogenome has unique gene order, with the genes split into two blocks encoded in opposite directions. The closest mitogenome available in GenBank belongs to the marine clam Mya arenaria (Myoida, Myidae). The average divergence of the sequences of these mitogenomes is quite high, in the range of 20%. Both mitogenomic and transcriptomic resources should prove very useful for elucidating population genetics and conservation issues involving this important species.

Complete female mitochondrial genome of Mytilus chilensis

Abstract The controversy surrounding the origin of antitropical distribution of Mytilus mussels and the taxonomic status of southern hemisphere populations remain unsolved, despite the efforts. One of the limiting factors remains the lack of the complete sequences of the representative mitochondrial genomes which would allow their proper comparison with the relatively well-represented northern hemisphere congeneric mussels. To fill this gap we sequenced the representative maternal (F) genome of a native Chilean mussel. The genome is 16,748bp long and structurally identical to the northern hemisphere M. edulis and M. galloprovincialis F genomes. However, the genetic distance from them (≈5%) is twice as high as the maximum distance between them (<2.5%). Thus, the notion that the name M. chilensis should be used for native Chilean Mytilus mussels, with the same rank as M. galloprovincialis and M. edulis is supported.

Disruption of doubly uniparental inheritance of mitochondrial DNA associated with hybridization area of European Mytilus edulis and Mytilus trossulus in Norway

Doubly uniparental inheritance of mitochondria (DUI) is best known in the blue mussel Mytilus. Under this model, two types of mitochondrial DNA exist: female type (F), transmitted from females to offspring of both genders, and male type (M), transmitted exclusively from males to sons. The mitogenomes are usually highly divergent, but an occasional replacement of a typical M genome by a particular F genome has been postulated to explain reduction of this divergence. Disruption of the DUI model has been reported in hybridization areas. Here, we present a new case of DUI disruption in a hybrid M. trossulus/M. edulis population from the North Sea (Norway). No M haplotypes derived from M. trossulus were identified in this population. Typical M haplotypes derived from M. edulis (ME) were rare. Two F-type haplogroups were found: one derived from M. edulis (FE) and the second derived from M. trossulus (FT). Many haplotypes from the FT group were recombinants, with the male CR sequence coming from the M. trossulus genome (FT1 haplogroup) in contrast to M. edulis CR as in the Baltic. FT1 haplotypes were abundant in the studied population, including homoplasmic females. However, males significantly more often carried these haplotypes; therefore, male heteroplasmy involved the original FE and recombinant FT, indicating that the FT genome undergoes masculinization. Structural similarity of FT1 CR with previously reported, masculinized Baltic haplotypes, which were derived from FE/ME recombination, provides further evidence that CR M–F recombination is a prerequisite for masculinization, also in the context of native M. trossulus mtDNA.

Complete male mitochondrial genomes of European Mytilus edulis mussels

Abstract Doubly uniparental inheritance (DUI) results in the existence of two gender-specific, divergent mtDNA lineages within a single species. Under DUI, the female genome (F) is transmitted from mothers to the whole offspring, and the male genome (M) is transmitted exclusively from fathers to sons. This system was first described in a marine mussels Mytilus edulis inhabiting European coastal waters, over a decade ago. Despite that, the complete sequence of the M genome from the European M. edulis mussels remained unknown. Here we announce it for the first time. The announcement is based on the two haplotypes isolated from heteroplasmic males of European M. edulis sampled at two moderately distant locations: southern North Sea and western Baltic. The two M genomes are quite similar both in length (16,631 and 16,632 bp) and in sequence (98.3%). Furthermore, both newly sequenced genomes are closely related to the genomes described from Baltic M. trossulus.

Recombinant mitochondrial genome with standard transmission route from Mediterranean mussel Mytilus galloprovincialis

Abstract Several bivalve species, including marine mussels Mytilus are atypical in having two gender-specific and highly divergent mtDNA genomes. This peculiar genetic system allows not only the recombination to occur but also facilitates its detection. Previous reports associated the existence of mosaic recombinant haplotypes with the switch of their transmission route. Here we report nearly complete sequence of a mitochondrial genome isolated from a homoplasmic female individual of Mediterranean Mytilus galloprovincialis. The genome has clear phylogenetic affinity with and organization identical to the M. galloprovincialis female haplotypes, in the coding part. However, the genome is very large, approximately 20,600 bp long, exclusively due to a long and complex control region. It contains an array of repeats, some of which are degenerated. A large part of the control region is derived from the paternal genome. This finding shows that not all haplotypes with recombinant control regions must be paternally inherited in Mytilus.

Actively transcribed and expressed atp8 gene in Mytilus edulis mussels

Background Animal mitochondrial genomes typically encode 37 genes: 13 proteins, 22 tRNAs and two rRNAs. However, many species represent exceptions to that rule. Bivalvia along with Nematoda and Platyhelminthes are often suspected to fully or partially lack the ATP synthase subunit 8 (atp8) gene. This raises the question as to whether they are really lacking this gene or is this maybe an annotation problem? Among bivalves, Mytilus edulis has been inferred to lack an ATP8 gene since the characterization of its mitochondrial genome in 1992. Even though recent bioinformatic analyses suggested that atp8 is present in Mytilus spp., due to high divergence in predicted amino acid sequences, the existence of a functional atp8 gene in this group remains controversial. Results Here we demonstrate that M. edulis mitochondrial open reading frames suggested to be atp8 (in male and female mtDNAs) are actively translated proteins. We also provide evidence that both proteins are an integral part of the ATP synthase complex based on in-gel detection of ATP synthase activity and two-dimensional Blue-Native and SDS polyacrylamide electrophoresis. Conclusion Many organisms (e.g., Bivalvia along with Nematoda and Platyhelminthes) are considered to be lacking certain mitochondrial genes often only based on poor similarity between protein coding gene sequences in genetically closed species. In some situations, this may lead to the inference that the ATP8 gene is absent, when it is in fact present, but highly divergent. This shows how important complementary role protein-based approaches, such as those in the present study, can provide to bioinformatic, genomic studies (i.e., ability to confirm the presence of a gene).

Mitogenomics of Perumytilus purpuratus (Bivalvia: Mytilidae) and its implications for doubly uniparental inheritance of mitochondria

Animal mitochondria are usually inherited through the maternal lineage. The exceptional system allowing fathers to transmit their mitochondria to the offspring exists in some bivalves. Its taxonomic spread is poorly understood and new mitogenomic data are needed to fill the gap. Here, we present for the first time the two divergent mitogenomes from Chilean mussel Perumytilus purpuratus. The existence of these sex-specific mitogenomes confirms that this species has the doubly uniparental inheritance (DUI) of mitochondria. The genetic distance between the two mitochondrial lineages in P. purpuratus is not only much bigger than in the Mytilus edulis species complex but also greater than the distance observed in Musculista senhousia, the only other DUI-positive member of the Mytilidae family for which both complete mitochondrial genomes were published to date. One additional, long ORF (open reading frame) is present exclusively in the maternal mitogenome of P. purpuratus. This ORF evolves under purifying selection, and will likely be a target for future DUI research.