Population Genetic Analyses Research Articles

Unaddressed hybridization between green (Bufotes viridis) and natterjack toads (Epidalea calamita) can lead to underestimation of genetic heterozygosity and inflated estimates of inbreeding

Abstract Hybridization is a major evolutionary process with significant consequences for the ecology and conservation of species and lineages. However, genetic studies that evaluate the level and influence of hybridization in wild populations of conservation concern remain scarce, despite recognition that unaddressed hybridization can bias results and lead to erroneous conclusions. Here, we explore how hybridization in mixed populations of the natterjack (Epidalea calamita) and green toads (Bufotes viridis) impacts the estimation of genetic parameters used to evaluate the conservation status of the regionally threatened natterjack toad. In order to determine the population structure of natterjack toads in the Central German Mining District near Leipzig (Germany) we have analysed 594 samples of bufonid eggs and tadpoles from eight different sites. Based on a detailed analysis of microsatellite loci, 392 individuals were determined as E. calamita, 158 as B. viridis and 44 as hybrids. Due to the high proportion of hybrids at some sites, estimates of inbreeding (FIS) in natterjack toads were biased towards higher values due to the detection of false homozygous individuals when hybrids were not removed from the dataset. Since inbreeding coefficients are a crucial parameter for population genetic analysis and biased estimates can give misleading results, our findings show that unaddressed hybridization can result in inflated estimates of inbreeding. This in turn, can lead to false conclusions that may ultimately misdirect conservation actions. We suggest that, if hybridization is documented for species pairs, it needs to be considered at an early stage when analysing populations.

Genetic Differentiation of Abies alba Outside Its Main Range Under Warm Meso- and Sub-Mediterranean Conditions in Italy and Switzerland.

Abies alba is an important European tree species currently mostly found at cool and humid sites in the montane zone. In the past, it grew under markedly warmer and drier climates during the Eemian and mid-Holocene, and cryptic Mediterranean populations confirm the species' capacity to grow under warm, summer-dry conditions. However, it is unknown if warm-loving Mediterranean occurrences are related to specific genetic properties (e.g., subspecies or ecotypes). Investigating the genetics of cryptic warm-loving populations is crucial for a better understanding of past and future population dynamics of A. alba. We genotyped 478 A. alba samples at 174 single-nucleotide polymorphisms (SNP), covering a broad latitudinal range from Southern Italy to Switzerland while accounting for local altitudinal gradients, and combined these newly introduced genotypes with those of other European Abies populations from the literature. Population genetic structure analyses grouped the warm-loving meso- and sub-Mediterranean populations into the same genetic cluster as the mountain populations of each region. The occurrence of three genetic clusters from Northern to Southern Italy is in line with the glacial refugia history. The inferred evolutionary and demographic history suggests a northward expansion of A. alba after glaciation, as well as a trans-Adriatic gene flow between Balkan and Southern Italian populations. Collectively, the combined genotypic data from individuals across the species' range demonstrate that cryptic Mediterranean populations of A. alba align with the local and large-scale genetic structure of populations from its main range, suggesting that the species is able to thrive in a warmer and drier environmental range than hitherto anticipated. This finding implies that it is unneeded to postulate extinct subspecies or ecotypes to explain the occurrence of meso- and sub-Mediterranean Eemian or mid-Holocene silver fir forests, with important implications for future A. alba population dynamics.

Genetic Variation in a Crossing Population of Camellia oleifera Based on ddRAD Sequencing and Analysis of Association with Fruit Traits

Tea oil is an important high-quality edible oil derived from woody plants. Camellia oleifera is the largest and most widely planted oil-producing plant in the Camellia genus in China, and its seeds are the most important source for obtaining tea oil. In current research, improving the yield and quality of tea oil is the main goal of oil tea genetic breeding. The aim of this study was to investigate the degree of genetic variation in an early crossing population of C. oleifera and identify single nucleotide polymorphisms (SNPs) and genes significantly associated with fruit traits, which can provide a basis for marker-assisted selection and gene editing for achieving trait improvement in the future. In this study, we selected a crossing population of approximately 40-year-old C. oleifera with a total of 330 samples. Then, ddRAD sequencing was used for SNP calling and population genetic analysis, and association analysis was performed on fruit traits measured repeatedly for two consecutive years. The research results indicate that over 8 million high-quality SNPs have been identified, but the vast majority of SNPs occur in intergenic regions. The nucleotide polymorphism of this population is at a low level, and Tajima’s D values are mostly greater than 0, indicating that the change in this population was not suitable for the model of central evolution. The population structure analysis shows that the population has seven theoretical sources of genetic material and can be divided into seven groups, and the clustering analysis results support the population structure analysis results. Association analysis identified significant SNPs associated with genes related to the seed number of a single fruit and seed kernel oil content. Our findings provide a basis for molecular breeding and future genetic improvement of cultivated oil tea.

High-throughput sequence-based microsatellite genotyping for the non-model Neotropical tree species Anadenanthera colubrina (Leguminosae)

Background and aims – Anadenanthera colubrina is a Neotropical native forest tree species with significant ecological importance in Seasonally Dry Tropical Forests. Developing genetic markers for this species is relevant for conservation, breeding, and evolutionary studies. Previously available genetic markers for A. colubrina consisted of a few microsatellites. Next-generation sequencing (NGS) strategies allow simple and cost-effective development of new SSR loci from low-coverage whole genome shotgun sequencing. The main aim was to develop microsatellite markers for sequence-based high-throughput genotyping (SSRseq) in the species and to characterize their information content against traditional capillary electrophoresis-based microsatellite data by estimating the amount of molecularly accessible size homoplasy of each locus. Additionally, the reliability of these markers for population genetic analysis was assessed by genotyping two age classes (reproductively mature trees and seedlings) in a typical location in Argentina. Key results – Sixty primer pairs targeting microsatellites were designed, of which 25 were validated with allelic error rates < 3% and genotype missingness < 20%. A significantly higher number of alleles per locus and heterozygosity was detected for SSRseq considering sequence polymorphisms compared to analysing the same data based on sequence size (length) only. Size homoplasy, calculated as the proportion of mismatches between datasets relative to the number of alleles differing in length, averaged 97.85% over all SSR loci. High levels of population genetic diversity were detected in adults and seedlings from Paranaense forests, exceeding those reported in previous studies of A. colubrina using traditional SSRs. The generated datasets increase the resolution of capillary-based microsatellite genotyping, allowing for more accurate inference of eco-evolutionary processes in non-model tree species.

Genetic characteristics of the bred group of deer and assessment of gene flow from it to natural populations

Here we present the results of genetic analysis of an artificial deer population, inhabiting an enclosed territory near Lida. The analysis was aimed at determining the precise origins and genetic characteristics of the population and disco vering presence of any gene flow between the artificial population and the wild deer of the region. We employed mitochondrial control region haplotypes analysis to identify origins and possible matrilineal hybrids, and hybrid classification of migration analysis based on microsatellite data to discover hybrid specimens and gene flow, respectively. We have determined that the artificial population in question belongs to the species Cervus canadensis sibiricus, or Altai wapiti, originates in the South Altai region, and possesses mediocre genetic diversity as can be expected from a population of this size. While singular results of hybrid analysis seem to indicate a possibility of rare interbreeding between escaped wapiti and wild deer, there are no indicators of any substantial gene flow from the artificial population into the wild, but we believe that it still warrants additional attention in order to prevent undesirable introgression.

АССОЦИАЦИЯ ПОЛИМОРФНЫХ ВАРИАНТОВ ГЕНА CAPN1 С ЖИВОЙ МАССОЙ У ОВЕЦ КАЛМЫЦКОЙ КУРДЮЧНОЙ ПОРОДЫ И ЕЕ ПОМЕСЕЙ С ПОРОДАМИ ШАРОЛЕ И ДОРПЕР

The purpose of research is to study the phenotypic effect of genotypes of the polymorphic calpain gene (CAPN1) on live weight in Kalmyk fat-tailed sheep and their crosses with the Charolais and Dorper breeds to substantiate the possibility of genetic marking of productivity at an early age. Objectives – carrying out genotyping and accounting of productivity indicators of animals of the Kalmyk fat-tailed breed (KK), Charolais (Sh), Dorper-Kalmyk crossbred ewes (½D×½KK), obtained purebred and crossbred young animals; establishing the influence of genotypes for the CAPN1 gene on live weight and average daily gain. It was found that in Kalmyk fat-tailed and Charolais breeding rams, carriers of the C allele in the CAPN1 gene were more often detected, while among purebred and crossbred ewes the occurrence of the C and T alleles was equal. In purebred young animals and ½ Sh ×½KK crosses, individuals with the T allele predominated; in ½ Sh ×¼D×¼KK crossbreeds, the carriers of the C allele predominated. Population genetic analysis in none of the groups did not reveal significant differences in the observed genotype frequencies from those theoretically expected at Hardy-Weinberg equilibrium. Regardless of breed and sex, the live weight of young animals at birth of different genotypes for the CAPN1 gene was similar, while at four months of age, both rams and lambs with the CC genotype had a significant superiority over their peers of other genotypes. In purebred rams, the greatest difference was observed between animals of the CC and ST genotypes, in crossbreeds ½ Sh ×½KK – animals of the CC and TT genotypes, which amounted to 10.4 and 8.1 %; 9.94 and 10.95 % (p < 0.05), respectively. Among the females, the greatest differences were observed during purebred breeding between animals of the CC and TT genotypes – 15.6 and 16.6 % (p < 0.05), respectively.

Whole-genome resequencing landscape of adaptive evolution in Relict gull (Larus relictus)

BackgroundThe relict gull (Larus relictus, Charadriiformes, Laridae) classified as vulnerable in the IUCN Red List is defined as a first-class national protected bird in China. However, our knowledge of the evolutionary history of L. relictus is limited. Here, we performed whole-genome resequencing of L. relictus (n = 14) and L. brunnicephalus (n = 3) to explore the genetic relationships and population structures and understand their adaptive evolution.ResultsThe whole genome resequencing generated 667.55 Gb clean reads with an average sequencing depth of ~ 29×. The genomic variant analysis identified 13,717,267 heterozygous SNPs in the samples obtained from 17 individuals. Population genetic diversity analysis revealed that low nucleotide diversity (0.00029) and no obvious population structure in L. relictus. Demographic history revealed that from 180 to 5 kya (thousand years ago), the effective population size (Ne) of L. relictus exhibited declines (24,000 to 5,000), with a very low range population size (2,200 to 5,000). In contrast, from 100 to 80 kya, L. brunnicephalus peaked in ancestral Ne, followed by distinct declines at ~ 70 kya (100,000 to 16,000). The findings identified several genes associated with the correlated changed life-history traits of L. relictus, including BMP4 involved in beak adaptation; HAND2, NEUROG1, COL11A2, and EDNRB involved in the evolution of the palate, soft palate, and tongue; PIGR and PLCB2 involved in an enhanced response to bitter taste by sensing chemical secretions released by staple food substrate insects to activate protective mechanisms. Furthermore, thirty-four genes related to sperm development and activity, including KLHL10 and TEKT3, were identified in the expanded gene family. In addition, MED1, CNOT9, NR5A1, and PATZ1 were involved in enhanced male hormone secretion and a high density of candidate genes associated with embryonic development were identified. The findings indicated that the L. relictus population was in a male-biased diffusion mode; the function of the TEKT3 gene showed that males played a dominant role in brooding, which enhanced their attraction to females. Our study revealed that significant enrichment of olfactory signaling pathway genes, including OR14C36, OR14J1, OR14I1, and OR14A16; inner ear development-related, including PTN, PTPN11, GATA2, ATP8B1, and MYO15A; and those related to hypoxic adaptation to high-altitude breeding and iris colour.ConclusionsBased on the results and the knowledge of this organism biology and habitat use, we infer that less adaptive evolutionary pressure on vision in L. relictus were related with their feeding behaviour and adaptation. In summary, this comprehensive analysis provides insights into the evolutionary features of L. relictus and a new perspective for scientific research on L. relictus to effectively determine its future survival viability.

QTL mapping and candidate gene analysis of element accumulation in rice grains via genome-wide association study and population genetic analysis

BackgroundToxic heavy metal elements in soils are major global environmental issues and easily migrate to crop grains to cause severe problems in human health, whereas moderately essential elements such as selenium are beneficial for human health. The accumulation of heavy metals and essential elements in rice grains and their genetic mechanisms are still poorly understood.ResultsWe conducted genetic dissection of four toxic heavy metal elements (lead, cadmium, mercury, and chromium), one quasi metallic element (arsenic), and one essential element (selenium) in grains of 290 Xian and 308 Geng rice accessions through a genome-wide association study (GWAS) based on three statistical models and assays of element concentrations from three environments. A total of 99 quantitative trait loci (QTLs) were identified. Among these QTLs, 18.2% overlapped between/among two or more elements, indicating that some QTLs related to the accumulation of certain elements may depend on other heavy metal elements or be involved in the collaborative transport of other elements. Moreover, at least 14 QTLs/regions were identified in the same regions, containing 12 cloned genes reported to be associated with element accumulation or tolerance-related traits, while the remaining 85 were new QTLs. A total of 62 promising candidate genes were identified from 50 major QTLs, of which 25 genes were newly discovered in this study. More importantly, population genetic analysis revealed 26 and 15 intraspecies divergent regions affecting element concentrations in the Xian and Geng subspecies, respectively, including 25 QTLs identified in this study and 13 previously reported and cloned genes.ConclusionsOur findings will facilitate further gene cloning and dissection of the genetic mechanisms of element accumulation in rice grains to improve grain quality.

Exploring the connection between climatic conditions and genetic properties of Abies alba Mill., including warm and summer-dry Mediterranean environments.

Abies alba Mill. is a prominent European tree species predominantly inhabiting cool and humid montane environments. However, paleoecological evidence reveals that during the Eemian and mid-Holocene, A. alba thrived in much warmer and drier climates. This capacity is nowadays reflected in cryptic meso- and sub-Mediterranean lowland populations. To link A. alba populations across diverse climates spanning from the Mediterranean lowlands to the Alpine timberline, we genotyped 421 specimens from Italy and Switzerland at 190 single-nucleotide polymorphisms (SNPs). Population genetic structure analyses indicate that isolated meso- and sub-Mediterranean lowland populations in Tuscany and Ticino align genetically with higher elevation populations in each region, suggesting that their capacity to thrive in warmer, drier conditions cannot be attributed to plantations with planting stock originating from different climates or to the occurrence of a single warm-adapted lineage showing a disjunct distribution, unless migration between Tuscany and Ticino stands occurred. Genotype-environment associations reveal that temperature seasonality, precipitation during critical seasons, and relative humidity are important for explaining the species' genetic variation. With genotype-environment and genotype-phenotype associations, we identified candidate adaptive genes potentially linked to climatic conditions and drought response. While certain adaptive alleles may have spread from Tuscany and Southern Italy or could be explained by a shared ancestry of Ticino and Tuscan populations, local adaptation may have occurred at specific loci. These findings underscore the importance of considering the hitherto overlooked lowland Mediterranean populations of A. alba to better understand the species' climatic niche and its potential for forest conservation and management under global warming.

Swatting Flies: Biting Insects as Non-Invasive Samplers for Mammalian Population Genomics.

Advances in next-generation sequencing have allowed the use of DNA obtained from unusual sources for wildlife studies. However, these samples have been used predominantly to sequence mitochondrial DNA for species identification while population genetics analyses have been rare. Since next-generation sequencing allows indiscriminate detection of all DNA fragments in a sample, technically it should be possible to sequence whole genomes of animals from environmental samples. Here we used a blood-feeding insect, tsetse fly, to target whole genome sequences of wild animals. Using pools of flies, we compared the ability to recover genomic data from hosts using the short-read sequencing (Illumina) and adaptive sampling of long-read data generated using Oxford nanopore technology (ONT). We found that most of the short-read data (85%-99%) was dominated by tsetse fly DNA and that adaptive sampling on the ONT platform did not substantially reduce this proportion. However, once tsetse reads were removed, the remaining data for both platforms tended to belong to the dominant host expected in the tsetse fly blood meal. Reads mapping to elephants, warthogs and giraffes were recovered more reliably than for buffalo, and there was high variance in the contribution of DNA by individual flies to the pools, suggesting that there are host specific biases. For elephants, using short-read sequencing we were able to identify over 300,000 unfiltered SNPs, which we used to estimate the allele frequencies and expected heterozygosity for the population. Overall, our results show that at least for certain wild mammals, it is possible to recover genome-wide host data from blood-feeding insects.

Population genetic analysis of clinical Mycobacterium abscessus complex strains in China.

To explore the genetic characteristics of the Mycobacterium abscessus complex (MABC) population in China, given its rising clinical importance among nontuberculous mycobacteria. We conducted population genetic analyses on 360 MABC genomes from China, focusing on core genome multilocus sequence typing (cgMLST), pan-genome characterization, population genetics, and antimicrobial resistance gene profiling. Our analysis identified 273 M. abscessus subsp. abscessus (MabA) and 87 M. abscessus subsp. massiliense (MabM) isolates, uncovering 68 sequence types (STs), with ST5 being the most common. cgMLST classified 33.3% of isolates into six dominant circulating clones (DCCs) and 49.4% into 59 genomic clusters at a threshold of 25 different alleles, including 18 international clusters linking Chinese isolates with seven other countries. The MABC pan-genome is open, with MabA exhibiting greater accessory gene diversity and higher gene turnover compared to MabM. Mobile genetic elements (MGEs), such as prophages and genomic islands, were prevalent across all genomes. 139 to 151 virulence factors (VFs) were identified per genome, with distinct accessory VFs in MabA and MabM affecting immune modulation and metabolism. Resistance gene profiling revealed ubiquitous mtrA, RbpA, and bla MAB, with MabA-specific erm(41) conferring resistance to macrolides and β-lactams. Common rrs and rrl gene mutations indicated widespread resistance to aminoglycosides and macrolides, while gyrA mutations suggested emerging fluoroquinolone resistance. An acquired erm(46) gene, likely obtained via phage-mediated horizontal gene transfer, was detected in one MabA strain. This study provides key genetic insights into the dynamics of MABC in China. The widespread distribution of DCCs, high genomic clustering rates, open pan-genome, and distinct resistance patterns between MabA and MabM, along with MGEs, highlight the need for targeted surveillance and tailored therapies to address emerging challenges in MABC infections.

Phylogenomics and species delimitation in the Lepidophyma sylvaticum complex (Squamata: Xantusiidae) using ddRADseq and morphological data.

The Lepidophyma sylvaticum complex occurs from west-central Nuevo León to the Sierra de Chiconquiaco in central Veracruz, Mexico. Morphological studies have revealed population groups that are "moderately divergent from each other" within the complex. In addition, a molecular phylogenetic study found L. sylvaticum to be strongly structured and paraphyletic with respect to L. micropholis. We used ddRADseq and morphological data to evaluate the phylogenetic relationships and species boundaries within the complex. Phylogenetic analyses performed using both concatenation and coalescent methods estimated the same well-supported tree, composed of five allopatric, strongly supported lineages of L. sylvaticum (from Nuevo León [NL]; Sierra de Álvarez, San Luis Potosí [ASLP]; southeastern San Luis Potosí, Querétaro, Hidalgo, and northwestern Veracruz [HSQV]; Tamaulipas [T]; and Puebla and northern Veracruz [PV]) branching off (in that order) from the base of the tree, with L. micropholis as the sister taxon to the last of them. A population genetics analysis revealed one pattern of genetic structure in L. micropholis and three in L. sylvaticum: one in the L. sylvaticum NL and ASLP lineages, another in the L. sylvaticum HSQV and T lineages, and a third in the L. sylvaticum PV lineage. The six lineages identified were genetically distinctive across our phylogenetic and population genetics analyses, and congruent with geography. Species tree and phylogenetic network analyses, considering the six lineages as potentially independent, inferred trees identical to the concatenated tree. Additionally, the phylogenetic network analysis detected a recent introgression event from the L. sylvaticum T lineage into L. micropholis. A time-calibrated tree indicated that the diversification of the complex began in the late Miocene and continued into the Pleistocene. A species delimitation analysis, based on the genealogical divergence index, suggested that the L. sylvaticum NL, ASLP, and PV lineages are independent evolutionary lineages. A morphological analysis showed that L. micropholis and the L. sylvaticum NL and ASLP lineages were distinguishable from each other and from the remaining L. sylvaticum lineages. We conclude that the L. sylvaticum NL and ASLP lineages, the two oldest in the complex, represent independent evolutionary lineages, and propose recognizing the other four lineages as subspecies (historical sublineages) of L. sylvaticum sensu stricto.

Comprehensive analysis of the multi-rings mitochondrial genome of Populus tomentosa

BackgroundPopulus tomentosa, known as Chinese white poplar, is indigenous and distributed across large areas of China, where it plays multiple important roles in forestry, agriculture, conservation, and urban horticulture. However, limited accessibility to the mitochondrial (mt) genome of P. tomentosa impedes phylogenetic and population genetic analyses and restricts functional gene research in Salicaceae family.ResultsSingle-molecule real-time (SMRT) sequencing technology was used to sequence, assemble, and annotate the mt genome of P. tomentosa. This genome has a complex structure composed of four circular molecules ranging from 153,004 to 330,873 base pairs (bp). Each of these four circular molecules contains unique gene sequences that constitute the mt genome of P. tomentosa. The mt genome comprises 69 functional genes, including 38 protein-coding genes (PCGs), 26 tRNA genes, and 5 rRNA genes. After removing duplications, 19 different tRNA coding genes remain, though only 10 amino acids can be recognized. The noncoding region constitutes 93.38% of the mt genome, comprising a large number of repetitive sequences, gene spacer regions, and insertion from chloroplast sequences. Specifically, 40 chloroplast-derived sequences, with a total length of 24,381 bp, were identified in P. tomentosa.ConclusionsIn the current study, the results provide mitochondrial genomic evidence for the maternal origin of P. tomentosa and enhance understanding of the gene dialog between organelle genomes, contributing to the conservation and utilization of the genetic resources of P. tomentosa.

Otolith chemistry suggests population heterogeneity within a genetically homogeneous Indian scad population along Indian coast

The Indian scad, Decapterus russelli is one of the most exploited pelagic resources of India. Population genetic analyses using mitochondrial and nuclear markers indicated a lack of genetic structuring among populations from Indian waters. As this species is highly migratory, it is also important to establish the environmental influence on its population structure. In the present study, the whole sagittal otolith composition was analysed using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) to assess spatial distribution of D. russelli collected from 4 sites along the Indian coast. Elemental ratios (Ba/Ca, Fe/Ca, K/Ca, Mg/Ca, Na/Ca, Sr/Ca and Zn/Ca) were analysed using univariate and multivariate statistics to determine whether this chemical signature can provide insight into the adaptive patterns. All element/Ca ratios are found to be significantly different between the four sampling locations (ANOVA, p < 0.05; MANOVA, p < 0.05). Five of the element/Ca ratios were found to be significantly different when the data was analysed coastwise (ANOVA, p < 0.05; MANOVA, p < 0.05). The LDA plot also showed the spatial heterogeneity of Element/Ca ratios between the four sampling sites, but some overlaps were also observed, reflecting the migratory ability of the species. This basic information on spatial ecology is required for formulating effective management and conservation strategies for the species.

Telomere-to-telomere sheep genome assembly identifies variants associated with wool fineness.

Ongoing efforts to improve sheep reference genome assemblies still leave many gaps and incomplete regions, resulting in a few common failures and errors in genomic studies. Here, we report a 2.85-Gb gap-free telomere-to-telomere genome of a ram (T2T-sheep1.0), including all autosomes and the X and Y chromosomes. This genome adds 220.05 Mb of previously unresolved regions and 754 new genes to the most updated reference assembly ARS-UI_Ramb_v3.0; it contains four types of repeat units (SatI, SatII, SatIII and CenY) in centromeric regions. T2T-sheep1.0 has a base accuracy of more than 99.999%, corrects several structural errors in previous reference assemblies and improves structural variant detection in repetitive sequences. Alignment of whole-genome short-read sequences of global domestic and wild sheep against T2T-sheep1.0 identifies 2,664,979 new single-nucleotide polymorphisms in previously unresolved regions, which improves the population genetic analyses and detection of selective signals for domestication (for example, ABCC4) and wool fineness (for example, FOXQ1).

Telomere-to-telomere genome and resequencing of 254 individuals reveal evolution, genomic footprints in Asian icefish, Protosalanx chinensis.

The Asian icefish, Protosalanx chinensis, has undergone extensive colonization in various waters across China for decades due to its ecological and physiological significance as well as its economic importance in the fishery resource. Here, we decoded a telomere-to-telomere (T2T) genome for P. chinensis combining PacBio HiFi long reads and ultra-long ONT (nanopore) reads and Hi-C data. The telomere was identified in both ends of the contig/chromosome. The expanded gene associated with circadian entrainment suggests that P. chinensis may exhibit a high sensitivity to photoperiod. The contracted genes' immune-related families and DNA repair associated with positive selection in P. chinensis suggested the selection pressure during adaptive evolution. The population genetic analysis reported the genetic diversity and genomic footprints in 254 individuals from 8 different locations. The natural seawater samples can be the highest diversity and different from other freshwater and introduced populations. The divergent regions' associated genes were found to be related to the osmotic pressure system, suggesting adaptations to alkalinity and salinity. Thus, the T2T genome and genetic variation can be valuable resources for genomic footprints in P. chinensis, shedding light on its evolution, comparative genomics, and the genetic differences between natural and introduced populations.

Identification and analysis of the genetic diversity of Dermacentor everestianus using mitochondrial 16S rRNA and COI sequences in the Xizang Autonomous Region of China

Ticks serve as vectors and reservoirs for a variety of pathogens, including protozoa, bacteria, and viruses, that cause tick-borne diseases in both humans and livestock. Despite their widespread distribution, their genetic diversity and population structure in Xizang have not been thoroughly investigated. In this study, sheep ticks were identified from four counties by microscopy, followed by genetic analysis of Dermacentor everestianus populations in Xizang based on mitochondrial 16S rRNA and cytochrome oxidase subunit 1 (COI) gene sequences. D. everestianus was the predominant species and 16S rRNA gene sequence analysis revealed nine haplotypes with seven nucleotide variations, showing a mean haplotype diversity (Hd) of 0.505 and mean nucleotide diversity (Pi) of 0.00129. Similarly, the COI sequences revealed 28 haplotypes with 26 nucleotide variations, yielding a mean Hd of 0.828 and a mean Pi of 0.00279. Molecular variance analysis (AMOVA) indicated that most genetic variation occurs within populations. In addition, neutral test analyses suggested potential bottleneck events in the D. everestianus populations. The haplotype network had a star-like distribution with the common haplotypes A1, A2, B1, and B5. The phylogenetic tree separated haplotypes of both genes into two branches on the haplotype network indicating no distinct geographical clustering. This study is the first to highlight the prevalence of D. everestianus in Xizang, revealing moderate genetic differentiation among the four populations, possibly due to host mobility. This intensive investigation of genetic variation and population structure contributes valuable insights into the current population genetics of D. everestianus in Xizang, China.

High genetic variation, low differentiation, and Pleistocene expansions of the migratory and endangered long-nosed tequila bat, Leptonycteris nivalis, inferred using both maternal and paternal genetic markers.

Tequila bats (genus Leptonycteris) have gained attention for their critical role in pollinating different plant species, especially Agave spp. and columnar cacti. Leptonycteris nivalis is the largest nectar-feeding bat in the Americas, and the females exhibit migratory behavior during the breeding season. Due to its relatively small and seemingly declining population sizes, this species is protected by government agencies in the United States and Mexico. We conducted population genetics and phylogeographic analyses to elucidate the genetic structure and demographic history of the species using two mitochondrial markers and a Y chromosome-associated gene, to describe both maternal and paternal lineages. We estimated high haplotypic diversity measures for the different markers (Dloop-Hd = 0.775; Cyt-b-Hd = 0.937; DBY -Hd = 0.946). We found that geographic genetic differentiation is very low, and there is high connectivity among localities. The estimated divergence time between L. nivalis and L. yerbabuenae, the other species in the genus found in Mexico, aligns with previous estimates for the genus (6.91-9.43 mya). A demographic expansion was detected approximately at 600 ka-700 ka (thousands of years ago). The historical demographic changes observed in L. nivalis appear to be associated with environmental shifts during the Pleistocene, which likely impacted the distribution range of the plants that these bats feed on, such as Agave species.

Genetic analysis of Pinna rudis L 1758 (Mollusca, Bivalvia, Pinnidae) in the Northwest Cabo Verde Islands (Central-East Atlantic).

The rough pen shell Pinna rudis Linnaeus, 1758 (family Pinnidae) is a mollusc with an Atlantic-Mediterranean distribution, typically inhabiting coarse sandy substrates. Habitat degradation is considered the primary cause of population decline, leading to the designation 'Vulnerable' in certain regions. In this study, we conducted a genetic analysis of populations of P. rudis from Cabo Verde and compared them with populations from the Mediterranean and Macaronesia. We based our analysis on two mitochondrial DNA markers, cytochrome oxidase I (COI) and 16S rRNA, and one nuclear marker, 28S rRNA. The results showed a strong genetic structuring among Macaronesia populations, with each island tending to have unique or exclusive haplotypes, though some sharing occurred between islands. We found significant genetic divergence between the population from Cabo Verde and the other sampled population, suggesting that P. rudis is not monotypic, but may include several cryptic species. Bayesian and maximum-likelihood phylogenetic analysis, including all Pinnidae species, indicated that P. rudis from Gorée Island (Senegal) and Baía das Gatas (São Vicente Island) might be undergoing speciation. The high genetic structure found for P. rudis could be influenced by hydrodynamic barriers, local currents and hydrographic isolation, in association with the short larval duration (planktotrophic) reported for this species. Altogether, our findings highlight significant genetic divergence in P. rudis populations, possibly supporting speciation events in the Cabo Verde archipelago among widely distributed taxonomic groups.

Population structure of Clinostomum complanatum (Trematoda: Digenea) with new data on haplotype diversity of flukes from Slovakia and Italy.

The fluke Clinostomum complanatum, a parasite of piscivorous birds, but also reptiles and rarely mammals, has established several foci in the western Palaearctic regions. Previous studies pointed out the complicated taxonomy of the genus, but broader population genetic analysis of C. complanatum has not yet been carried out. The aim of this study was to determine the structure, intraspecific variability, and diversity of mitochondrial cox1 haplotypes of C. complanatum from different localities in Slovakia (Danube floodplain forests) and Italy (Emilia-Romagna and Tuscany), as well as to evaluate the interrelationships among populations from Europe, the Middle East, and North Africa. The genetic structure of C. complanatum from Slovakia and Italy was represented by a great number of haplotypes, showing stable populations with high intraspecific diversity. The haplotypes of samples from other localities (Romania, Turkey, Egypt, and Iran) showed possible gene flow among the populations from Central Europe down to the Mediterranean region, North Africa, and the Middle East. The genetic homogeneity of these samples can be linked to the distribution and migratory routes of the definitive hosts, aquatic piscivorous birds, mainly herons and cormorants, that spread parasite eggs among the continents.