mtDNA Loci Research Articles

BIOLOGICAL NOMENCLATURE (TAXONOMY) AND CLASSIFICATION OF HONEY BEE. CURRENT STATE AND PROBLEMS

The chronology of scientific research achievements in the process of determining the biological nomenclature and classification of honey bees and the implementation of the results obtained for use in the practice of beekeeping in the selection and preservation of aboriginal breeds (races) of bees in modern environmental conditions is highlighted. An attempt is made to focus this study in the field of analysis of the chronology of methods used in the historical aspect of determining the taxonomy of insects, in particular bees. At the same time, the problems and prospects of scientific research in modern economic and natural and climatic conditions of the development of the industry are reflected. It is shown that initially only morphometry was used worldwide to identify bee breeds. However, morphometric features are not always informative in identifying subspecies, since they are subject to variability under the influence of environmental conditions. Later, biochemical methods for identifying bee subspecies based on polymorphism of allozyme loci were developed. It is shown that at the same time, methods for identifying bee subspecies based on polymorphism of mitochondrial DNA (mtDNA) loci were developed. This polymorphism was successfully used in phylogenetic and phylogeographic studies of honey bees. The disadvantage of mtDNA markers is the exclusively maternal type of inheritance. At the same time, methods for identifying bee subspecies were developed taking into account polymorphism of nuclear DNA (nDNA) loci. Recently, methods for identifying bee subspecies based on SNP analysis have been developed. These markers have become widely used in population, evolutionary and phylogenetic studies of bees due to the development of next-generation sequencing methods NGS (Next Generation Sequencing) Illumina. SNP markers are characterized by high resolution due to their number and stable inheritance over several generations, which can be successfully used in genetic mapping, population and evolutionary studies, selection of lines for economically useful traits and disease resistance, identification of taxonomic affiliation of bee families.

Genetic consequences of the dramatic decline in the north-west pre-caspian saiga (Saiga tatarica tatarica) population: a comparison of modern and museum samples on mtDNA and microsatellite loci

The North-West Pre-Caspian saiga (Saiga tatarica tatarica) population numbers was about 800 thousand individuals in the middle of the last century. There was a dramatic decline at the end of the 20th century, and by 2015 this saiga population numbers and was about five thousand animals. Our paper presents the results of microsatellite loci and of the mtDNA control region analysis obtained for the museum saiga samples from the North-West Pre-Caspian population, collected at the peak of their numbers in the 1950s. We compared these data with our previous results for the samples collected during the population depression in1999–2016. There were no noticeable differences of mtDNA control region diversity between the museum and modern saiga samples. The most vivid genetic consequence of the severe decline in population numbers was a significant increase of the inbreeding coefficient (Fis) calculated from the microsatellite loci.

Genetic Consequences of the Dramatic Decline in the Northwest Pre-Caspian Saiga (Saiga tatarica tatarica) Population: A Comparison of Modern and Museum Samples on mtDNA and Microsatellite Loci

Genetic Consequences of the Dramatic Decline in the Northwest Pre-Caspian Saiga (Saiga tatarica tatarica) Population: A Comparison of Modern and Museum Samples on mtDNA and Microsatellite Loci

Association between mitochondrial DNA genotype and sperm motility in humans

The relationship between genetic alterations in mitochondrial DNA (mtDNA) and progressive motility (PR) and rapid progressive motility (grade A) of ejaculated human spermatozoa remains unclear. In this study, we explored the association between human mtDNA genotype and sperm PR and grade A by analyzing mtDNA copy number, loci, haplogroup, rearrangement, deletions, and duplications and sperm motility parameters. Human sperm mtDNA copy number, loci and haplogroups were not associated with human sperm motility PR or A grade. However, the cumulative frequency of human sperm mtDNA rearrangements (including deletions and duplications) in participants with high PR and grade A ratio was higher than in participants with low PR and grade A ratio. Additional studies are needed to understand the relationship between mtDNA genotypes, including deletions and duplications, and human sperm motility.

Unravelling the maternal evolutionary history of the African leopard (Panthera pardus pardus).

The African leopard (Panthera pardus pardus) has lost a significant proportion of its historical range, notably in north-western Africa and South Africa. Recent studies have explored the genetic diversity and population structure of African leopards across the continent. A notable genetic observation is the presence of two divergent mitochondrial lineages, PAR-I and PAR-II. Both lineages appeared to be distributed widely, with PAR-II frequently found in southern Africa. Until now, no study has attempted to date the emergence of either lineage, assess haplotype distribution, or explore their evolutionary histories in any detail. To investigate these underappreciated questions, we compiled the largest and most geographically representative leopard data set of the mitochondrial NADH-5 gene to date. We combined samples (n = 33) collected in an altitudinal transect across the Mpumalanga province of South Africa, where two populations of leopard are known to be in genetic contact, with previously published sequences of African leopard (n = 211). We estimate that the maternal PAR-I and PAR-II lineages diverged approximately 0.7051 (0.4477-0.9632) million years ago (Ma). Through spatial and demographic analyses, we show that while PAR-I underwent a mid-Pleistocene population expansion resulting in several closely related haplotypes with little geographic structure across much of its range, PAR-II remained at constant size and may even have declined slightly in the last 0.1 Ma. The higher genetic drift experienced within PAR-II drove a greater degree of structure with little haplotype sharing and unique haplotypes in central Africa, the Cape, KwaZulu-Natal and the South African Highveld. The phylogeographic structure of PAR-II, with its increasing frequency southward and its exclusive occurrence in south-eastern South Africa, suggests that this lineage may have been isolated in South Africa during the mid-Pleistocene. This hypothesis is supported by historical changes in paleoclimate that promoted intense aridification around the Limpopo Basin between 1.0-0.6 Ma, potentially reducing gene flow and promoting genetic drift. Interestingly, we ascertained that the two nuclear DNA populations identified by a previous study as East and West Mpumalanga correspond to PAR-I and PAR-II, respectively, and that they have come into secondary contact in the Lowveld region of South Africa. Our results suggest a subdivision of African leopard mtDNA into two clades, with one occurring almost exclusively in South Africa, and we identify the potential environmental drivers of this observed structure. We caution that our results are based on a single mtDNA locus, but it nevertheless provides a hypothesis that can be further tested with a dense sample of nuclear DNA data, preferably whole genomes. If our interpretation holds true, it would provide the first genetic explanation for the smaller observed size of leopards at the southernmost end of their range in Africa.

Molecular characterisation of Pinus sylvestris (L.) in Ireland at the western limit of the species distribution

BackgroundScots pine (Pinus sylvestris L.) underwent significant population declines across much of northwest Europe during the mid-to-late Holocene and was thought to have become extirpated in Ireland from about 400 AD. However, most extant populations are plantations reintroduced from Scotland. Others are naturalised therefrom and one in Western Ireland is a putative relict. In this paper, Scots pine in Ireland are genetically described for the first time.ResultsUsing two mitochondrial (mtDNA) loci, eight chloroplast (cpSSR) and 18 nuclear (nSSR) loci, the genetic composition and diversity of 19 Irish Scots pine populations is described and compared to other European populations. All trees sampled in Ireland were fixed for mitotype a, which is the most common across northwest Europe. By contrast, cpSSR (HCP = 0.967) and nSSR (He = 0.540) variation was high, and comparable with estimates for other regions across the species range. Differentiation at both sets of loci were similarly low (cpSSR FST = 0.019; nSSR FST = 0.018), but populations from continental Europe were significantly differentiated from all Irish populations based on nSSR variation.ConclusionsAll Irish Scots pine are likely part of a common Irish-Scottish gene pool which diverged from continental Scots pine following post-glacial recolonisation. A high genetic diversity and an absence of evidence of inbreeding suggests the regional decline of Scots pine did not critically reduce allelic variation. The post-glacial relationship between Irish and Scottish pine is discussed, and a suggestion from recent palaeoecological work that reintroduced Scots pine be managed as a native species is now further supported by genetic data.

Low Genetic Variability of the Tundra Vole in Lithuania.

The distribution and spread of the tundra vole (Alexandromys oeconomus) in Lithuania have been documented over the last 70 years, but the genetic diversity of the species has not been studied. In this study, we examined A. oeconomus trapped in three sites in northern and western Lithuania using mtDNA sequence analysis of the cytb and control region. The western and northern sites are separated by anthropogenic landscape barriers. The western site is subject to regular spring flooding. Phylogenetic analyses of the studied individuals placed them in the Central European phylogroup, suggesting that Lithuanian A. oeconomus originated from northeastern Poland. In Lithuania, the genetic diversity of A. oeconomus at both mtDNA loci was relatively low (Hd < 0.6, π < 0.002) compared to that found in other European samples (Hd = 0.833-0.958; π = 0.00402-0.01552). Individuals analyzed in Lithuania were genetically different from samples collected in Poland and Northern Europe (ΦST > 0.15, p < 0.05). The genetic divergence between the western and northern samples of A. oeconomus in Lithuania, together with the low genetic variability among the voles studied, provides new insights into the phylogeography of the species and the influence of barriers on the colonization of the country.

A Molecular Phylogeny of Stylodipus (Dipodidae, Mammalia): A Small Genus with a Complex History

A range-wide phylogenetic/phylogeographic study of the three-toed jerboas of the genus Stylodipus is conducted using the mitochondrial cytb gene and fragments of several nuclear genes. The genus has been believed to include three species: S. telum (W Central Asia, SE Europe), S. andrewsi (E Central Asia), and S. sungorus (Dzungar basin). Our data support the dichotomy between S. andrewsi and the other taxa forming S. telum species group. Within the latter, both mtDNA and nuclear loci indicate a species-level divergence between S. telum and the S. t. birulae lineage (Zaisan depression, NE Kazakhstan), previously considered a subspecies of S. telum and here elevated to full species. S. sungorus is recovered as a close sister group to S. birulae on the basis of nuclear data but clustered with S. telum in the mitochondrial tree. The latter taxon is the most variable and includes two closely related eastern and western sublineages, separated by the Volga-Ural sands and joined by a more divergent S. t. karelini lineage (E Kazakhstan). The observed mitonuclear discordance is hypothesized to occur due to mtDNA introgression resulting from hybridization between S. sungorus and S. t. karelini, which highlights the important role of reticulations in the evolution of Dipodidae.

Genetic features of bivalve transmissible neoplasia in blue mussels from the Kola Bay (Barents Sea) suggest a recent trans-Arctic migration of the cancer lineages.

Ecology and biogeography of bivalve transmissible neoplasia (BTN) are underexplored due to its recent discovery and a challenging diagnostics. Blue mussels harbour two evolutionary lineages of BTN, MtrBTN1 and MtrBTN2, both derived from Mytilus trossulus. MtrBTN1 has been found only in M. trossulus from North Pacific. MtrBTN2 parasitizes different Mytilus spp. worldwide. BTN in M. trossulus in the Atlantic sector has never been studied. We looked for BTN in mussels from the Barents Sea using flow cytometry of cells, qPCR with primers specific to cancer-associated alleles and sequencing of mtDNA and nuclear loci. Both MtrBTN1 and MtrBTN2 were present in our material, though their prevalence was low (~0.4%). All cancers parasitized M. trossulus except one, MtrBTN1, which was found in a hybrid between M. trossulus and M. edulis. The mtDNA haplotypes found in both lineages were nearly identical to those known from the Northwest Pacific but not from elsewhere. Our results suggest that these two lineages may have arrived in the Barents Sea in recent decades with the maritime transport along the Northern Sea Route. A young evolutionary age of MtrBTN1 seems to indicate that it is an emerging disease in the process of niche expansion. Comparing the new and the published sequence data on tumour suppressor p53, we proved that the prevalence of BTN in mussels can reach epizootic levels. The finding of diverse recombinants between paternally and maternally inherited mtDNAs in somatic tissues of M. trossulus was an unexpected result of our study.

Quantifying impacts of an environmental intervention using environmental DNA.

Environmental laws around the world require some version of an environmental-impact assessment surrounding construction projects and other discrete instances of human development. Information requirements for these assessments vary by jurisdiction, but nearly all require an analysis of the biological elements of ecosystems. Amplicon-sequencing-also called metabarcoding-of environmental DNA (eDNA) has made it possible to sample and amplify the genetic material of many species present in those environments, providing a tractable, powerful, and increasingly common way of doing environmental-impact analysis for development projects. Here, we analyze an 18-month time series of water samples taken before, during, and after two culvert removals in a salmonid-bearing freshwater stream. We also sampled multiple control streams to develop a robust background expectation against which to evaluate the impact of this discrete environmental intervention in the treatment stream. We generate calibrated, quantitative metabarcoding data from amplifying the 12s MiFish mtDNA locus and complementary species-specific quantitative PCR data to yield multispecies estimates of absolute eDNA concentrations across time, creeks, and sampling stations. We then use a linear mixed effects model to reveal patterns of eDNA concentrations over time, and to estimate the effects of the culvert removal on salmonids in the treatment creek. We focus our analysis on four common salmonid species: cutthroat trout (Oncorhynchus clarkii), coho salmon (Oncorhynchus kisutch), rainbow trout (Oncorhynchus mykiss), and sockeye salmon (Oncorhynchus nerka). We find that one culvert in the treatment creek seemed to have no impact while the second culvert had a large impact on fish passage. The construction itself seemed to have only transient effects on salmonid species during the two construction events. In the context of billions of dollars of court-mandated road culvert replacements taking place in Washington State, USA, our results suggest that culvert replacement can be conducted with only minimal impact of construction to key species of management concern. Furthermore, eDNA methods can be an effective and efficient approach for monitoring hundreds of culverts to prioritize culverts that are required to be replaced. More broadly, we demonstrate a rigorous, quantitative method for environmental-impact reporting using eDNA that is widely applicable in environments worldwide.

Molecular and acoustic evidence support the species status of Anthus rubescens rubescens and Anthus [rubescens] japonicus (Passeriformes: Motacillidae).

The Buff-bellied Pipit Anthus rubescens comprises two allopatric subspecies groups: A. r. rubescens and A. r. alticola in North America and A. [r.] japonicus in north-east Asia. Despite their great morphological resemblance in breeding plumage, most individuals can be assigned to one or the other subspecies group in non-breeding plumage. Allopatric distributions, morphological differentiation and previously reported molecular divergence suggested the need for additional taxonomic study to assess the rank of these two populations. To resolve the taxonomy of the Buff-bellied Pipit species complex we analysed i) two mitochondrial DNA (mtDNA) loci and ii) nine bioacoustic parameters across 69 sound recordings (338 flight calls) recovered from public databases using principal component analysis and Euclidean distance measures. By comparing our mtDNA and call divergence measures with similar values measured between long-recognised species pairs of the genus, we show that the level of mitochondrial and acoustic divergence between the two Buff-bellied Pipit subspecies groups is typical of species-level divergence in the genus Anthus. Therefore, we recommend splitting the Buff-bellied Pipit species complex into two species: Anthus rubescens (American Pipit) and Anthus japonicus (Siberian Pipit). Our results also suggest that the Water Pipit A. spinoletta deserves taxonomic reassessment as its lineages are highly divergent in acoustics and mtDNA, while mtDNA relationships suggest paraphyly relative to the Rock Pipit A. petrosus. Our work highlights the crucial importance of integrative approaches in taxonomy and the usefulness of bioacoustics in studying cryptic diversity.

Genetic Characteristics of the Baikal Seal Pusa sibirica Population.

Nucleotide sequence diversity in two mtDNA loci (the cytochrome b gene and the control region) was for the first time studied in the Baikal seal Pusa sibirica with the use of several spatiotemporal samples. The population was found to be evolutionarily young and to be in the stage of demographic expansion.

Characterising a genetic stronghold amidst pervasive admixture: Morelet’s crocodiles (Crocodylus moreletii) in central Yucatan

When backcrosses are fertile, interbreeding between endangered taxa can lead to the admixture of gene pools under threat. One such case pertains to the Mesoamerican crocodile Crocodylus moreletii, a species which shows strong signatures of both recent hybridisation and historic intogression with the American crocodile C. acutus across large parts of its range. In the present paper, we use RAD-seq derived SNPs (4980 nuclear and seven mtDNA loci) to demonstrate that C. moreletii populations inhabiting the region of Calakmul in central Yucatan (Mexico) are rather unaffected by hybridization, despite being surrounded by coastal areas where pervasive admixture has previously been documented. All (based on fastSTRUCTURE) and 96% (based on NGSadmix) of 84 genotyped individuals from 18 sampled waterbodies (locally termed aguadas) were free from nuclear introgression of C. acutus DNA at at threshold of 0.95. Seven individuals (8%) possessed a C. acutus mtDNA haplotype, five of which were derived from two adjacent, rather peripheral aguadas. Spatial inferences based on a DAPC and fineRADstructure further showed that the region of Calakmul is inhabited by three genetic clusters spanning across a set of distinct aguadas each. Taken together, our findings reveal that central Yucatan contains the currently largest documented stronghold of C. moreletii populations only marginally affected by introgression, which has major implications for the conservation management of this important flagship species.

Genome-wide analysis reveals distinct global populations of pink bollworm (Pectinophora gossypiella)

The pink bollworm (Pectinophora gossypiella) is one of the world’s most destructive pests of cotton. This invasive lepidopteran occurs in nearly all cotton-growing countries. Its presence in the Ord Valley of North West Australia poses a potential threat to the expanding cotton industry there. To assess this threat and better understand population structure of pink bollworm, we analysed genomic data from individuals collected in the field from North West Australia, India, and Pakistan, as well as from four laboratory colonies that originated in the United States. We identified single nucleotide polymorphisms (SNPs) using a reduced-representation, genotyping-by-sequencing technique (DArTseq). The final filtered dataset included 6355 SNPs and 88 individual genomes that clustered into five groups: Australia, India-Pakistan, and three groups from the United States. We also analysed sequences from Genbank for mitochondrial DNA (mtDNA) locus cytochrome c oxidase I (COI) for pink bollworm from six countries. We found low genetic diversity within populations and high differentiation between populations from different continents. The high genetic differentiation between Australia and the other populations and colonies sampled in this study reduces concerns about gene flow to North West Australia, particularly from populations in India and Pakistan that have evolved resistance to transgenic insecticidal cotton. We attribute the observed population structure to pink bollworm’s narrow host plant range and limited dispersal between continents.

Genetic methods in honey bee breeding.

The honey bee Apis mellifera is a rather difficult object for selection due to the peculiarities of its biology. Breeding activities in beekeeping are aimed at obtaining bee colonies with high rates of economically useful traits, such as productivity, resistance to low temperatures and diseases, hygienic behavior, oviposition of the queen, etc. With two apiaries specializing in the breeding of A. m. mellifera and A. m. carnica as examples, the application of genetic methods in the selection of honey bees is considered. The first stage of the work was subspecies identification based on the analysis of the polymorphism of the intergenic mtDNA locus tRNAleu-COII (or COI-COII) and microsatellite nuclear DNA loci Ap243, 4a110, A24, A8, A43, A113, A88, Ap049, A28. This analysis confirmed that the studied colonies correspond to the declared subspecies. In the apiary with A. m. mellifera, hybrid colonies have been identified. A method based on the analysis of polymorphisms of the tRNAleu-COII locus and microsatellite nuclear DNA loci has been developed to identify the dark forest bee A. m. mellifera and does not allow one to differentiate subspecies from C (A. m. carnica and A. m. ligustica) and O (A. m. caucasica) evolutionary lineages from each other. The second stage was the assessment of the allelic diversity of the csd gene. In the apiary containing colonies of A. m. mellifera (N = 15), 20 csd alleles were identified. In the apiary containing colonies of A. m. carnica (N = 44), 41 alleles were identified. Six alleles are shared by both apiaries. DNA diagnostics of bee diseases showed that the studied colonies are healthy. Based on the data obtained, a scheme was developed for obtaining primary material for honey bee breeding, which can subsequently be subjected to selection according to economically useful traits. In addition, the annual assessment of the allelic diversity of the csd gene will shed light on the frequency of formation of new allelic variants and other issues related to the evolution of this gene.

GENETIC CHARACTERISTICS OF THE BAIKAL SEAL POPULATION (&lt;i&gt;PUSA SIBIRICA&lt;/i&gt;)

For the first time, the diversity of nucleotide sequences of different mtDNA loci (the cytochrome b gene and the control region) of the Baikal seal (Pusa sibirica) based on of several spatio-temporal samples was studied. It is shown that in the evolutionary time scale the population formed recently and is in the stage of demographic expansion.

Movement and Genomic Methods Reveal Mechanisms Promoting Connectivity in a Declining Shorebird: The Lesser Yellowlegs

Integrating tracking technology and molecular approaches provides a comprehensive picture of contemporary and evolutionary mechanisms promoting connectivity. We used mitochondrial DNA and double digest restriction-site associated DNA (ddRAD) sequencing combined with satellite telemetry to investigate the connectivity of geographically disparate breeding populations of a declining boreal shorebird, the lesser yellowlegs (Tringa flavipes). We were able to track 33 individuals on their round-trip migrations to Central and South America and back to the boreal wetlands of North America. Nearly all (93%) adults captured on the breeding grounds returned to within 5 km of the original capture site, with a median dispersal distance of 629 m. While our telemetry data revealed limited breeding dispersal in adults, genetic data uncovered significant interconnectedness across the species’ range. Very little genetic structure was estimated at ddRAD autosomal (ΦST = 0.001), Z-linked (ΦST = 0.001), and mtDNA loci (ΦST = 0.020), and maximum likelihood-based clustering methods placed all individuals in a single cluster regardless of capture location, indicating the species is panmictic. Our data indicate that large-scale juvenile dispersal is the main mechanism maintaining connectivity in this species, resulting in the absence of genomic structure.

A revision of the West African freshwater crab genus Afrithelphusa Bott, 1969 (Brachyura: Deckeniidae: Deckeniinae) based on new morphological and genetic data

ABSTRACT The freshwater crab genus Afrithelphusa Bott, 1969, is endemic to Sierra Leone and Guinea and comprises four species: A. afzelii Colosi, 1924, A. gerhildae Bott, 1969, A. leonensis (Cumberlidge, 1987), and A. monodosa (Bott, 1959). Afrithelphusa taxonomy is reviewed based on type material and newly obtained undescribed specimens. The species are redescribed, based on new morphological and genetic data, their amended diagnostic features are provided, and the generic diagnosis is updated. An identification key is provided to distinguish between the four Afrithelphusa species, and a tree of phylogenetic relationships based on two mtDNA loci (COI and 16S RNA) is provided, which supports the continued recognition of this genus and its assigned species.

DddA homolog search and engineering expand sequence compatibility of mitochondrial base editing

Expanding mitochondrial base editing tools with broad sequence compatibility is of high need for both research and therapeutic purposes. In this study, we identify a DddA homolog from Simiaoa sunii (Ddd_Ss) which can efficiently deaminate cytosine in DC context in double-stranded DNA (dsDNA). We successfully develop Ddd_Ss-derived cytosine base editors (DdCBE_Ss) and introduce mutations at multiple mitochondrial DNA (mtDNA) loci including disease-associated mtDNA mutations in previously inaccessible GC context. Finally, by introducing a single amino acid substitution from Ddd_Ss, we successfully improve the activity and sequence compatibility of DdCBE derived from DddA of Burkholderia cenocepacia (DdCBE_Bc). Our study expands mtDNA editing tool boxes and provides resources for further screening and engineering dsDNA base editors for biological and therapeutic applications.

Pseudophilautus pardus, a junior synonym of P. viridis (Amphibia: Anura: Rhacophoridae).

Pseudophilautus pardus, was first described in 2007, based on a single specimen collected in Sri Lanka during the 19th century. Its absence in recent surveys suggested that the species was extinct. The distinctive spotted dorsal coloration, together with other morphological features, was used to distinguish it from congeners. In 2013, we discovered a specimen resembling the holotype of P. pardus in Samanala Strict Nature Reserve. A DNA analysis based on the 16S rRNA mt-DNA locus recovered a low uncorrected pairwise genetic distance of 0.77-0.96% between the recent example of 'P. pardus' and a congener, P. viridis. Molecular species delimitation methods suggest P. pardus and P. virids to be a single operational taxonomic unit. Morphological analyses using the freshly collected specimen, together with numerous photographic records show P. viridis to be a highly polychromatic species within which the coloration observed in P. pardus too occurs, though rarely. We conclude that P. pardus is a junior synonym of P. viridis.