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The Eurasian perch Perca fluviatilis (Linnaeus, 1758) is a common species of fish in northern ecosystems. The species demonstrates high phenotypic diversity when it inhabits various types of water bodies. Here, we investigate whether there is a relationship between the genetic variability and phenotypic diversity in natural perch populations. Perca fluviatilis samples (n = 218) were collected from seven localities in the Ob-Irtysh river basin in western Siberia, Russia. We used color morphs and standard morphometric approach to study phenotypic diversity, allozyme and ISSR-PCR markers to study the genetic variability of the perch. In total, 19 types of perch colouration were found. The number of color morphs varied from 4 to 16 in different reservoirs. The sets of colour morphs and prevailing coloration types, as well as some morphometric characteristics, were significantly different in all studied populations. Low allozyme variability was identified in the perch. The average observed and expected allozyme heterozygosity was 0.003 and 0.056, respectively; 13% of the loci were polymorphic. The genetic diversity (h) of the markers (ISSR) was 0.31; from 53% to 96% of the bands were polymorphic. Genetic differentiation in the perch was high, especially in allozyme loci. The FST and GST values were 0.39 and 0.085 for allozyme and ISSR markers, respectively. The genetic variability indices of the perch did not correlate with phenotypic diversity. Our results suggest that the use of different phenotypic or genetic markers can provide extremely different information on the level of variability in the population., The study was supported by the Government of the Tyumen Region within the framework of the project of the West Siberian Interregional Scientific and Educational Center No. 89-DON (2). The authors have declared that no competing inter-ests exist. We are grateful to V. Evdash and N. Zhuravleva, specialists of the Center for Academic Writing "Impulse", for English editing.

This review summarizes the development of population genetics and population genomics studies of forest trees in Slovakia during the past 40 years. Various protein and DNA markers have been applied during this period to address several topics in evolutionary genetics and biogeography of trees: allozymes, uniparentally inherited chloroplast and mitochondrial markers, simple sequence repeats and single nucleotide polymorphisms. The main object of studies of phylogeny and postglacial migration were Fagus sylvatica s.l. and eastern-Mediterranean firs (Abies Mill. section Abies), where the divergence of genetic lineages (species and subspecific taxa) in time, as well as colonization of the current ranges during the Holocene were reconstructed. The studies on intraspecific gene flow and homoploid hybridization focused on hybrid swarms Pinus sylvestris/P. mugo and firs. Unusual maternal inheritance of chloroplast DNA was revealed in P. mugo × P. sylvestris crosses. Contrasting geographical structures of hybrid zones were revealed in wind-dispersed vs. animal-dispersed trees. Within the studies of adaptation, signals of selection were identified both in field observations and common-garden experiments on Picea abies, F. sylvatica and A. alba. Perspectives of ongoing research employing next-generation sequencing were shortly outlined.

The Ural Mountains and the West Eurasian Taiga forests are one of the most important centers of genetic diversity for Larix sibirica Ledeb. Forest fragmentation negatively impacts forest ecosystems, especially due to the impact of their intensive use on the effects of climate change. For the preservation and rational use of forest genetic resources, it is necessary to carefully investigate the genetic diversity of the main forest-forming plant species. The Larix genus species are among the most widespread woody plants in the world. The Siberian larch (Larix sibirica, Pinaceae) is found in the forest, forest-tundra, tundra (Southern part), and forest-steppe zones of the North, Northeast, and partly East of the European part of Russia and in Western and Eastern Siberia; in the Urals, the Siberian larch is distributed fragmentarily. In this study, eight pairs of simple sequence repeat (SSR) primers were used to analyse the genetic diversity and population structure of 15 Siberian larch populations in the Urals. Natural populations in the Urals exhibit indicators of genetic diversity comparable to those of Siberia populations (expected heterozygosity, He = 0.623; expected number of alleles, Ne = 4017; observed heterozygosity, Ho = 0.461). Genetic structure analysis revealed that the examined populations are relatively highly differentiated (Fst = 0.089). Using various algorithms for determining the spatial genetic structure, the examined populations formed three groups according to geographical location. The data obtained are required for the development of species conservation and restoration programs, which are especially important in the Middle Urals, which is the region with strong forest fragmentation.

Cryptic and hybridizing species may lack diagnostic taxonomic characters leaving researchers with semi-diagnostic ones. Identification based on such characters is probabilistic, the probability of correct identification depending on the species composition in a mixed population. Here we test the possibilities of applying a semi-diagnostic conchological character for distinguishing two cryptic species of blue mussels, Mytilus edulis and M. trossulus. These ecologically, stratigraphically and economically important molluscs co-occur and hybridize in many areas of the North Atlantic and the neighboring Arctic. Any cues for distinguishing them in sympatry without genotyping would save much research effort. Recently these species have been shown to statistically differ in the White Sea, where a simple character of the shell was used to distinguish two mussel morphotypes. In this paper, we analyzed the associations between morphotypes and species-specific genotypes based on an abundant material from the waters of the Kola Peninsula (White Sea, Barents Sea) and a more limited material from Norway, the Baltic Sea, Scotland and the Gulf of Maine. The performance of the “morphotype test” for species identification was formally evaluated using approaches from evidence-based medicine. Interspecific differences in the morphotype frequencies were ubiquitous and unidirectional, but their scale varied geographically (from 75% in the White Sea to 15% in the Baltic Sea). In addition, salinity-related variation of this character within M. edulis was revealed in the Arctic Barents Sea. For every studied region, we established relationships between the proportions of the morphotypes in the populations as well as between the proportions of the morphotypes in samples and the probabilities of mussels of different morphotypes being M. trossulus and M. edulis. We provide recommendations for the application of the morphotype test to mussels from unstudied contact zones and note that they may apply equally well to other taxa identified by semi-diagnostic traits.

The large-scale hybridization of fishes of the subfamily Leuciscinae in the subordinate systems of the Dnieper river basin is presented by data on two pair of species, Rutilus rutilus and Abramis brama, Alburnus alburnus and Scardinius erythrophthalmus. The hybridization and occurrence of F1 hybrids are confirmed with allozyme spectra and morphological characters on series of samples. The presence of single individuals sharply deviating in morphological characters among the bleak and common rudd hybrids suggests limited backcrossing with rudd. A complete morphometric analysis of the characteristics of bleak and rudd hybrids has been carried out. The paper discusses the issue of intergeneric hybridization in nature, which is a unique phenomenon characteristic only of some groups of cyprinids. The most likely reason is the overestimation of the taxonomic status of European cyprinids, which is confirmed by the insignificant level of intergeneric genetic divergence.

It is important to know the plant reproduction mode for the exploration and management of thenatural plant population, as well as for breeding program and germoplasm regeneration of accessions. Bromus auleticus Trinius is a perennial winter grass, native of South America which shows high potential for forage and has good adaptation to soil and climate adversities. The objective of this work was to characterize the reproductivesystem of the species and to estimate its crossing frequency among the accesses existing at Epagri/ Experiment Station of Lages. Four progenies originated from natural crosses were characterized by ten allozymic loci (PGI, SKDH, IDH, PRX1, PRX2, PRX3, MDH1, MDH3, 6PGD and EST). Crossing frequency of unilocus and multilocuswere estimated using the allelic frequencies and the MLT program (Ritland, 1990). The unilocus estimation variedfrom 0,001 to 1,648 and had a mean frequency of 0,571. The multilocus had a frequency of 1,096. Based on these results, the reproduction mode is predominantly allogamic. O conhecimento do modo de reprodução das espécies vegetais é fundamental na exploração e no manejo de populações naturais, nos programas de melhoramento genético e regeneração de acessos em bancos de germoplasmas. Bromus auleticus Trinius é uma gramínea hibernal, perene, nativa da América do Sul, com alto potencial forrageiro e adaptativo a condições adversas de clima e solo. O objetivo do trabalho foi caracterizar osistema reprodutivo da espécie e estimar a taxa de cruzamento entre acessos existentes na Epagri/Estação Experimental de Lages. Quatro progênies oriundas de cruzamentos naturais foram caracterizadas utilizando-sedez locos isoenzimáticos (PGI, SKDH, IDH, PRX1, PRX2, PRX3, MDH1, MDH3, 6PGDH, £-EST). Foram estimadas as taxas de cruzamento uniloco e multilocos, usando as freqüências alélicas das progênies com análise do programaMLT (Ritland, 1990). As estimativas uniloco variaram de 0,001 a 1,648, com média de 0,571, e a multilocos foi de 1,096, o que caracteriza que o modo de reprodução predominante é a alogamia.

Within the genus Centaurea L., polyploidy is very common, and it is believed that, as to all angiosperms, it was key in the history of its diversification and evolution. Centaurea tentudaica is a hexaploid from subsect. Chamaecyanus of unknown origin. In this study, we examined the possible autopolyploid or allopolyploid origin using allozymes and sequences of three molecular markers: nuclear-ribosomic region ETS, and low-copy genes AGT1 and PgiC. We also included three species geographically and morphologically close to C. tentudaica: C. amblensis, C. galianoi, and C. ornata. Neighbor-Net and Bayesian analyses show a close relationship between C. amblensis and C. tentudaica and no relationship to any of the other species, which suggest that C. tentudaica is an autopolyploid of C. amblensis. Allozyme banding pattern also supports the autopolyploidy hypothesis and shows high levels of genetic diversity in the polyploid, which could suggest multiple origins by recurrent crosses of tetraploid and diploid cytotypes of C. amblensis. Environmental niche modeling was used to analyze the distribution of the possible parental species during the present, Last Glacial Maximum (LGM), Last Interglacial Period (LIG), and Penultimate Glacial Maximum (PGM) environmental conditions. Supporting the molecular suggestions that C. tentudaica originated from C. amblensis, environmental niche modeling confirms that past distribution of C. amblensis overlapped with the distribution of C. tentudaica., This research was funded by the Catalan government (“Ajuts a grups consolidats” 2017-SGR1116)., 1. Introduction 2. Materials and Methods 2.1. Sampling Strategy and Markers 2.2. DNA Amplification, Cloning, and Sequencing 2.3. Sequence Alignment and Phylogenetic Analyses 2.4. Allozyme Extraction and Analyses 2.5. Ecological Niche Modeling 3. Results 3.1. ETS Region 3.2. Low-copy Genes AGT1 and PgiC 3.3. Allozymes 3.4. Ecological Niche Modeling 4. Discussion 4.1. Relative Utility of ETS, AGT1, and PgiC in Disentangling the Origin of Centaurea tentudaica 4.2. The Origin of Centaurea tentudaica 5. Conclusions

Identification of discrete stocks of Greenland halibut is an important aspect of proper fisheries exploitation. Available literature data indicated a lack of significant inter‑area differences between Greenland halibut populations from the Northeast Atlantic. To define the population diversity, two aspects were taken into account: enzyme‑genetic diversity and the concentration of heavy metals in tissues. Seven allozyme loci variations were used to characterize the genetic structure of four populations of Greenland halibut from the Western Barents Sea region. The samples were collected from the spawning area in the period when this species took migration to spawn. The sample RH4 was significantly different from the other samples collected from the same location (RH2 and RH3) and depth for over two days. Another sample (RH8), collected from the nearby area 6 days later was similar to the samples RH2, RH3. We noticed a significant divergence between the sample RH4 and the three remaining samples, where the value of the index FST fluctuated about 0.40 and approximately 0.01 between three similar populations. This genetic fluctuation negates the thesis of a panmictic character of the Western Atlantic population. Feeding groups of Greenland halibut are moving along the Barents Sea shelf and they are exposed to different heavy metals concentrations according to the food preferences or the exact place of feeding. We identified similar concentrations of heavy metals, i.e., Zn, Cu, Cd, and Pb in all samples. Trace metal analysis of aquatic organisms from the Barents Sea can provide important information on the degree of environmental contamination, and the potential impact of seafood consumption.

A recent study argued that coalescent-based models of species delimitation mostly delineate population structure, not species, and called for the validation of candidate species using biological information additional to the genetic information, such as phenotypic or ecological data. Here, we introduce a framework to interrogate genomic datasets and coalescent-based species trees for the presence of candidate species in situations where additional biological data are unavailable, unobtainable or uninformative. For de novo genomic studies of species boundaries, we propose six steps: (1) visualize genetic affinities among individuals to identify both discrete and admixed genetic groups from first principles and to hold aside individuals involved in contemporary admixture for independent consideration; (2) apply phylogenetic techniques to identify lineages; (3) assess diagnosability of those lineages as potential candidate species; (4) interpret the diagnosable lineages in a geographical context (sympatry, parapatry, allopatry); (5) assess significance of difference or trends in the context of sampling intensity; and (6) adopt a holistic approach to available evidence to inform decisions on species status in the difficult cases of allopatry. We adopt this approach to distinguish candidate species from within-species lineages for a widespread species complex of Australian freshwater fishes (Retropinna spp.). Our framework addresses two cornerstone issues in systematics that are often not discussed explicitly in genomic species discovery: diagnosability and how to determine it, and what criteria should be used to decide whether diagnosable lineages are conspecific or represent different species.

Molecular markers prove to be an invaluable tool in assessing the introduction dynamics, pattern of range expansion, and population genetics of an invasive species. Ventenata dubia (Leers) Coss. (Aveneae; ventenata) is a diploid, primarily self-pollinating, annual grass native to Eurasia and Northern Africa. The grass has a detailed herbarium collection history in the western United States since its discovery in eastern Washington in 1952. Genetic analysis of 51 invasive populations (1636 individuals) of V. dubia, coupled with historical records, suggests moderate propagule pressure from multiple introductions, followed by local or regional range expansion. Allozyme analysisdetected nine multilocus genotypes (MLGs) across eight western US states. A single MLG, referred to as the most common genotype, was detected in 37 of 51 (72.5%) invasive populations across all states. The other eight MLGs were generally found in fewer populations, with limited geographic distributions. Despite multiple introductions, invasive populations exhibit low levels of genetic admixture, low levels of genetic diversity within populations (A = 1.03, %P = 2.94, Hexp = 0.007) and high genetic differentiation among populations (GST= 0.864). The apparent reduced evolutionary potential of most V. dubia populations did not preclude the initial establishment and rapid spread of this species across its new range in the western US.

Habitat fragmentation is a serious threat to species survival. In Vietnam, Cycas balansae has been considered as threatened species because of the reduction and fragmentation of its habitats and over-exploitation. We assessed genetic variability and the pattern of population structure among six populations sampled in four provinces: Hoa Binh, Ha Nam, Ninh Binh and Quang Ninh. Polyacrylamide gel electrophoresis was performed on leaf tissues from 152 individuals representing 6 populations of C. balansae. Six of twelve enzyme systems were used to estimate genetic diversity at population and species levels. Eleven loci were examined. The allozyme data showed high levels of genetic diversity within all populations, ranging from 0.538 in Ba Sao to 0.628 in Tan Dan (average 0.576). The maintenance of high levels of expected heterozygosity (average 0.571) and low in observed heterozygosity (average 0.347) might be related to great heterozygote deficiency and increased frequencies of rare alleles. Genetic differentiation among populations was low (Dst = 0.036 and Gst = 0.064), indicating high level of gene flow (Nm = 3.22). Isolation by geographical distance was observed, however, no significant relationship between genetic distances and geographical distances was recorded. Our studies suggest small population sizes of cycads brought about by fragmentation of its habitats, over-exploitation, and increasing number of inbred individuals within populations.

Genetic variation and differentiation in taxa of the genus Viola L. sect. Viola, subsections Viola and Rostratae (Kupffer) W. Becker, were studied from natural populations occurring in Iran. Two isoenzyme systems, Phosphoglucoisomerase (PGI) and Aconitase (ACO), encoding four putative loci, were employed to detect interspecific and intraspecific genetic variation. Considering the patterns of isoenzyme variation in the studied taxa, it is evident that V. caspia (Rupr.) Freyn subsp. caspia and V. capia subsp. sylvestroides Marcussen are closely related. The species V. alba Bess. subsp. alba and V. sintenisii W. Becker are isoenzymatically well characterized as distinct genetic entities.

U radu je izvršen integrativno-taksonomski pristup analize kompleksa vrsta Merodon avidus (Diptera: Syrphidae), na geografski i vemenski obimnom materijalu. U okviru genetičke karakterizacije kriptičnih vrsta navedenog kompleksa, bazirane na 3' i 5'fragmentima mitohondrijalnog COI gena, određeni su parametri genetičke varijabilnosti i utvrđeni jedinstveni i deljeni haplotipovi u okviru i između pretpostavljenih vrsta. Rezultati genetičke varijabilnosti COI DNK sekvenci pokazali su da jedinke sa ostrva Krf i Evia i poluostrva Peloponez, preliminarno identifikovane kao M. moenium, predstavljaju novu, endemsku vrstu proučavanogkompleksa. Dijagnostički enzimski lokusi pokazali su da M. avidus i M. moenium predstavljaju sestrinske vrste, koje su se, u okviru kompleksa vrsta M. avidus, poslednje razdvojile. Zaključeno je da se kompleks vrsta M. avidus sastoji od 5 vrsta: M. avidus, M. moenium, M. megavidus, M. ibericus i M. aff. moenium. U nastavku, oslanjajući se na moderne tehnike veštačke inteligencije, izvršeno je modelovanje distribucije vrsta i poređenje sličnosti utvrđenih ekoloških niša. U poslednjem segmentu istraživanja, prednosti veštačke inteligencije iskorišćene su u modelovanju sistema za determinaciju jedinki sestrinskih vrsta u uzorku, na osnovu adekvatne varijable.Ovo istraživanje doprinelo je karakterizaciji biodiverziteta osolikih muva, rasvetljavanju taksonomskog statusa vrsta i kreiranju smernica za definisanje budućih konzervacionih programa zaštite biodiverziteta vrsta Merodon avidus kompleksa., During this research, an integrative-taxonomic analysis of M. avidus species complex was performed. The study was based on geographically and temporally extensive material. Genetic characterisation of cryptic species, based on 5’ and 3’ regions of COI gene, defined parameters of genetic variability. Shared and unique haplotypes between and within of cryptic species were detected. The results of genetic variability analysis based on COI gene showed that specimens from the islands Corfu, Evia and half-island Peloponnese, which were preliminarily identified as M. moenium, represent a new, endemic species of the selected complex. Diagnostic enzyme loci showed that M. avidus and M. moenium represent sibling species, which were the last one who separated within M. avidus complex. According to current information, it was concluded that M. avidus complex consists of 5 species: M. avidus, M. moenium, M. megavidus, M. ibericus and M. aff. moenium. In the next chapter, relying on modern techniques of artificial intelligence, the species distribution modelling and the comparison of ecological niches were performed. In the last part of the research, the advantages of artificial intelligence were used in order to model a system that was able to determinate one of two sibling species, based on appropriate predictor. This research has generally contributed to a characterization of hoverfly diversity and helped resolving a taxonomic status of species in one of the most challenging groups in Syrphidae family. Genetic differentiation data represent directions for defining future conservation strategies for biodiversity protection of defined cryptic species of Merodon avidus complex

U radu je izvršen integrativno-taksonomski pristup analize kompleksa vrsta Merodon avidus (Diptera: Syrphidae), na geografski i vemenski obimnom materijalu. U okviru genetičke karakterizacije kriptičnih vrsta navedenog kompleksa, bazirane na 3' i 5'fragmentima mitohondrijalnog COI gena, određeni su parametri genetičke varijabilnosti i utvrđeni jedinstveni i deljeni haplotipovi u okviru i između pretpostavljenih vrsta. Rezultati genetičke varijabilnosti COI DNK sekvenci pokazali su da jedinke sa ostrva Krf i Evia i poluostrva Peloponez, preliminarno identifikovane kao M. moenium, predstavljaju novu, endemsku vrstu proučavanogkompleksa. Dijagnostički enzimski lokusi pokazali su da M. avidus i M. moenium predstavljaju sestrinske vrste, koje su se, u okviru kompleksa vrsta M. avidus, poslednje razdvojile. Zaključeno je da se kompleks vrsta M. avidus sastoji od 5 vrsta: M. avidus, M. moenium, M. megavidus, M. ibericus i M. aff. moenium. U nastavku, oslanjajući se na moderne tehnike veštačke inteligencije, izvršeno je modelovanje distribucije vrsta i poređenje sličnosti utvrđenih ekoloških niša. U poslednjem segmentu istraživanja, prednosti veštačke inteligencije iskorišćene su u modelovanju sistema za determinaciju jedinki sestrinskih vrsta u uzorku, na osnovu adekvatne varijable.Ovo istraživanje doprinelo je karakterizaciji biodiverziteta osolikih muva, rasvetljavanju taksonomskog statusa vrsta i kreiranju smernica za definisanje budućih konzervacionih programa zaštite biodiverziteta vrsta Merodon avidus kompleksa., During this research, an integrative-taxonomic analysis of M. avidus species complex was performed. The study was based on geographically and temporally extensive material. Genetic characterisation of cryptic species, based on 5’ and 3’ regions of COI gene, defined parameters of genetic variability. Shared and unique haplotypes between and within of cryptic species were detected. The results of genetic variability analysis based on COI gene showed that specimens from the islands Corfu, Evia and half-island Peloponnese, which were preliminarily identified as M. moenium, represent a new, endemic species of the selected complex. Diagnostic enzyme loci showed that M. avidus and M. moenium represent sibling species, which were the last one who separated within M. avidus complex. According to current information, it was concluded that M. avidus complex consists of 5 species: M. avidus, M. moenium, M. megavidus, M. ibericus and M. aff. moenium. In the next chapter, relying on modern techniques of artificial intelligence, the species distribution modelling and the comparison of ecological niches were performed. In the last part of the research, the advantages of artificial intelligence were used in order to model a system that was able to determinate one of two sibling species, based on appropriate predictor. This research has generally contributed to a characterization of hoverfly diversity and helped resolving a taxonomic status of species in one of the most challenging groups in Syrphidae family. Genetic differentiation data represent directions for defining future conservation strategies for biodiversity protection of defined cryptic species of Merodon avidus complex

Resumo Podocarpus lambertii (Podocarpaceae) é conhecido popularmente como pinheiro-bravo. Ocorre em várias formações florestais da Mata Atlântica, principalmente na Floresta Ombrófila Mista. Devido à intensa exploração madeireira sofrida pela espécie, ela está presente na categoria “Quase ameaçada” da Lista Vermelha das Espécies Ameaçadas da IUCN. Diante disso, este estudo teve por objetivo caracterizar a diversidade e estrutura genética de 12 populações no Estado de Santa Catarina, a fim de gerar informações para fundamentar estratégias de conservação para a espécie. Foram coletadas folhas de 50 indivíduos adultos em cada população. Para acessar os níveis e a distribuição da diversidade genética, utilizaram-se 10 sistemas enzimáticos. As frequências alélicas, a porcentagem de locos polimórficos (P100%), as heterozigosidades observada (HO) e esperada (HE), o índice de fixação (f) e a divergência genética (FST) foram calculados com auxílio do programa FSTAT. Os 10 sistemas enzimáticos analisados revelaram 11 locos com um total de 32 alelos (A= 1,8). As populações de Podocarpus lambertii apresentaram baixa diversidade genética (P100% = 52,7; HO = 0,049; HE = 0,079). As frequências alélicas das populações apresentaram desvios significativos das esperadas em Equilíbrio de Hardy-Weinberg, evidenciando um deficit de heterozigotos (f = 0,374). A presença de alelos raros nas populações e uma divergência genética significativa (FST = 0,303) evidenciam um baixo fluxo gênico histórico e um grande risco de perda de diversidade. Os resultados obtidos indicam a necessidade de conservação in situ das várias populações de Podocarpus lambertii e de um aumento da conectividade entre as populações remanescentes. Abstract Podocarpus lambertii (Podocarpaceae) is popularly known in Brazil as pinheiro-bravo. It occurs in several forest formations of the Atlantic Rain Forest, mainly in the Ombrophilous Mixed Forest. Due to the high wood exploitation suffered by the species, it is listed as ‘Almost Threatened’ in The IUCN Red List of Threatened Species. In face of this, the objective of the present study was to characterize the genetic diversity and structure of 12 natural populations of Podocarpus lambertii in Santa Catarina state, aiming to generate information for the definition of effective conservation strategies for the species. To access genetic diversity levels and distribution, it was used 10 allozymic systems. Along with the assistance of the software Fstat, the allelic frequencies (A), polymorphic loci percentage (P), observed heterozigosity (Ho), expected heterozigosity, fixation index (f) and genetic divergence between populations (FST) were calculated. In 11 loci 32 alleles (A= 1,8) were detected. A low genetic diversity was detected for Podocarpus lambertii populations (P100% = 52,7; H O = 0,049; H E = 0,079). The mean fixation index obtained (f = 0,374) indicates a deficit of heterozygous individuals. A high number of low frequency alleles was detected and together with the high genetic divergence (FST = 0,303), it points towards the necessity of in situ conservation of a large number of Podocarpus lambertii populations and the need to increase the connectivity among the remnant populations.

Amphibians are the most vulnerable vertebrates to biodiversity loss mediated by habitat destruction, climate change and diseases. Informed conservation management requires improving the taxonomy of anurans to assess reliably the species’ geographic range. The genusOdontophrynusthat is geographically refined to Argentina, Bolivia, Brazil, Uruguay and Paraguay includes currently 12 nominal species with many populations of uncertain taxonomic assignment and subsequently unclear geographic ranges. In this study, we applied integrative taxonomic methods combining molecular (mitochondrial 16S gene), allozyme, morphological and bioacoustic data to delimit species of the genusOdontophrynussampled from throughout Argentina where most species occur. The combined evidence demonstrates one case of cryptic diversity and another of overestimation of species richness. The populations referred to asO. americanuscomprise at least three species. In contrast,O. achalensisandO. barrioirepresent junior synonyms of the phenotypically plastic speciesO. occidentalis. We conclude that each of the four species occurring in Argentina inhabits medium to large areas. The Red List classification is currently “Least Concern”. We also propose a phylogenetic hypothesis for the genus and associated generaMacrogenioglottusandProceratophrys(Odontophrynidae).

The taxonomic rank of the tiny-leaved terrestrial orchid Cephalanthera subaphylla Miyabe & Kudô has been somewhat controversial, as it has been treated as a species or as an infraspecific taxon, under C. erecta (Thunb.) Blume [C. erecta var. subaphylla (Miyabe & Kudô) Ohwi and C. erecta f. subaphylla (Miyabe & Kudô) M. Hiro]. Allozyme markers, traditionally employed for delimiting species boundaries, are used here to gain information for determining the taxonomic status of C. subaphylla. To do this, we sampled three populations of five taxa (a total of 15 populations) of Cephalanthera native to the Korean Peninsula [C. erecta, C. falcata (Thunb.) Blume, C. longibracteata Blume, C. longifolia (L.) Fritsch, and C. subaphylla]. Among 20 putative loci resolved, three were monomorphic (Dia-2, Pgi-1, and Tpi-1) across the five species. Apart from C. longibracteata, there was no allozyme variation within the remaining four species. Of the 51 alleles harbored by these 17 polymorphic loci, each of the 27 alleles at 14 loci was unique to a single species. Accordingly, we found low average values of Nei’s genetic identities (I) between ten species pairs (from I = 0.250 for C. erecta versus C. longifolia to I = 0.603 for C. falcata vs. C. longibracteata), with C. subaphylla being genetically clearly differentiated from the other species (from I = 0.349 for C. subaphylla vs. C. longifolia to 0.400 for C. subaphylla vs. C. falcata). These results clearly indicate that C. subaphylla is not genetically related to any of the other taxa of Cephalanthera that are native to the Korean Peninsula, including C. erecta. In a principal coordinate analysis (PCoA), C. subaphylla was positioned distant not only from C. falcata, C. longibracteata, and C. longifolia, but also from C. erecta. Finally, K = 5 was the best clustering scheme using a Bayesian approach, with five clusters precisely corresponding to the five taxa. Thus, our allozyme results strongly suggest that C. subaphylla merits the rank of species., This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2013R1A1A2063524 to MYC and NRF-2013R1A1A3010892 and NRF-2017R1A2B4012215 to MGC) and was carried out as part of the Infrastructure for the Conservation and Restoration of Rare and Endemic Plants in Korea National Arboretum that supported to MGC from 2015 to 2019.

Protection and management of closely related endangered species and subspecies at a very narrow regional scale is the origin of multiple dysfunctional conservation decisions. These include artificially increased IUCN risk assessment categories and derived consequences: poor effectiveness in allocating public and private funds or repeat of unnecessary actions/facilities. Data provided by the revisited study of a group of W Mediterranean larkspurs (Delphinium ser. Fissa), including new data on demography, niche modeling, genetic diversity and phylogeography, contributed to a new and wider analysis of causes of threat. Although current IUCN Red List regulations did not allow for assessments at levels higher than a specific rank, scientific information suggests that in some cases this could be a better approach for sound scientifically-based biodiversity conservation planning and action., Funded by Diputació de Barcelona, Servei d'Espais Naturals (Contract FBG 307118); Generalitat de Catalunya (GReB-AGAUR, Grant 2017SGR-1116) and Universitat de Barcelona (Ajut CRAI Accés Obert - UB 2019).

Neutral genetic diversity found in plant species usually leaves an indelible footprint of historical events. Korea’s main mountain range (referred to as the Baekdudaegan [BDDG]), is known to have served as a glacial refugium primarily for the boreal and temperate flora of northeastern Asia. In addition, life-history traits (life forms, geographic range, and breeding systems) influence the within- and among-population genetic diversity of seed plant species. For example, selfing species harbor significantly less within-population genetic variation than that of predominantly outcrossers. A previous study of two Liparis species (L. makinoana and L. kumokiri) emphasizes the role of the abovementioned factors shaping the levels of genetic diversity. Liparis makinoana, mainly occurring on the BDDG and self-incompatible, harbors high levels of within-population genetic diversity (expected heterozygosity, HeP = 0.319), whereas there is no allozyme variation (HeP = 0.000) in L. kumokiri, which is self-compatible and mainly occurs in lowland hilly areas. To determine if this trend is also found in other congeners, we sampled five populations of L. krameri from the southern part of the Korean Peninsula and investigated the allozyme-based genetic diversity at 15 putative loci. The somewhat intermediate levels of within-population genetic variation (HeP = 0.145) found in L. krameri are most likely due to its occurrence in mountainous areas that, despite being outside of the main ridge of the BDDG, still served as refugia, and a self-incompatible breeding system. Management strategies are suggested for L. krameri and L. makinoana based on the levels and distribution of genetic diversity and inbreeding., This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2013R1A1A2063524 to MYC and NRF-2013R1A1A3010892 and NRF-2017R1A2B4012215 to MGC) and was carried out as part of the Infrastructure for the Conservation and Restoration of Rare and Endemic Plants in Korea National Arboretum that supported to MGC from 2015 to 2019.

Main results of allozyme-geographical study of the polymorphism, differentiation and origin of natural Pinus sylvestris L. populations in 6phylogeographical regions – Middle Siberian Tableland, Lena-Angara Tableland, South Siberia Mountains, Near-Baikalia, Western Trans-Baikalia and Northern Mongolia – have been briefly analyzed and generalized. A relative homogeneity of the population polymorphism in these regions has been revealed, with the exception of marginal ones (Tura, Ulan-Bator). Genetic distances of M. Nei (1978, DN78) between the extremely remote populations in the investigated regions reach the level of the geographical race, but they are several time lesser in the regions’ limits – not more of the genosystmatic level of the middle differentiated local populations (DN78 = 0.010–0.012). The geographical group of the South Siberia Mountains stand apart distinctly (DN78 = 0.018±0.004) in the genogeographical structure of P. sylvestris on the basis of Nei`s genetic distances analysis and of the genosystematic scale (Sannikov, Petrova, 2012). Besides the Selenga population group (in the rank of local population, DN78 = 0.013), also as well the Angara-Yenisei and North-Baikal population group (in the rank of subpopulation, DN78 = 0.004–0.008) stand apart on the rest of the territory. Gradient-analysis of the genetic distances revealed the borders between the phylogeographical groups of pine populations on the Lena-Angara and Central-Siberian plateaus, on the one hand, in the South Siberia Mountains, Near-Baikal and Trans-Baikalia, on the other hand, and an important role of l. Baikal as a barrier of the migration and differentiation of the populations. As an analysis result of minimal genetic distances between 25 P. sylvestris populations in the south «non-glacial» zone and the group of 8 populations in the «glacial» zone 4 main refugiums (South-Near-Baikal, South-Ural, Upper-Vitim and Upper-Olekma) and several secondary refugiums have been revealed.

Background. The population structure of relict and protected species Cepaea vindobonensis Fer. (Mollusca, Gastropoda, Pulmonata) in a north-eastern part of the modern area has been studied. State population gene pools (11 populations) for environmental activities need to be assessed. Materials and Methods. Using allozymes (esterases) and DNA markers (RAPD, ISSR) analysis was performed. The methods polyacrylamide gel electrophoresis and the DNA in the agarose gel were used. Results. The populations with a low level of heterozygosity and high heterozygosity were identified. Group located in the east, did not differ in allelic diversity of the western groups. The average expected heterozygosity for allozyme He = 0,217 ± 0,080 and DNA markers He = 0,164 ± 0,024. However, the population of the Austrian snail in the study area is strongly separated from each other (Fst = 0,231, Gst = 0,265, Фst = 0,261-0,263, Nm = 0,630-0,832). The effective population size (Ne), calculated on the basis of the inbreeding coefficient, was level 0,868 ± 0,028 of the total population. The effective population size, calculated on the basis of the linear regression equation between genetic and geographic distance (Ne = 0,36-2,6) and calculated by a factor of subdivision of populations (Ne = 6,3 ± 0,1), in comparison with the effective size of the populations of common species of terrestrial mollusks was the same. Conclusion. Thus, on the basis of these data, in general, we can say, the Austrian population snails have a higher potential for life. However, the reduction of allelic diversity, the increase in the coefficient of inbreeding in some groups and a high level of isolation of populations of C. vindobonensis in the north-eastern part of the range is a concern and requires an adjustment in respect of environmental protection measures of this species.

The genetic polymorphism in selectively reared in Bulgaria, local honey bee populations of Apis mellifera macedonica subspecies (type rodopica), has been studied, using analysis of six enzyme systems (MDH-1, ME, EST-3, ALP, PGM and HK) corresponding to 6 loci. Totally 458 worker bees from 12 bee breeding bases for artificially inseminated queens were used for this study. All these stations are part of the National Bee Breeding Association which officially implements a National Program for sustainable beekeeping in Bulgaria. All of the six loci were found to be polymorphic. Only EST-3 locus was established as fixed in one of the investigated populations. Polymorphism with three alleles was ascertained for MDH, ME, ALP, PGM and HK loci and with five alleles for EST-3 locus. The most common alleles in almost all of the populations were MDH-1 100, ME 100, EST-3 100, PGM 100 and HK 100. Two private alleles (frequency < 0.05) were found for two of the populations. The calculated level of polymorphism was 88.33% in only one of the populations and 100% - in all others. The observed and expected heterozygosities (Ho and He) ranged from 0.157 to 0.250 and from 0.206 to 0.272, respectively. The estimated mean FST value from allozyme data was 0.035. On the bases of the allele frequencies of the studied allozyme loci the Nei's (1972) genetic distance was estimated. It ranged between 0.002 and 0.060 among the populations studied.

Paracentrotus gaimardi is a sea urchin reaching a maximum of 42 mm of diameter. It presents five morphotypes in sympatry characterized by their distinctive spine colors (e.g. green, black, gray, brown and rose). P. gaimardi is considered as a vulnerable species and it is included on the list of endangered species of the Brazilian coastline. In this work, allozyme electrophoresis was used to studied the genetic variation among the five color morphotypes, from two different geographic locations along the coast of Rio de Janeiro State: Itaipu and Prainha, both in sympatry and allopatry. The underlying null hypothesis is that the different spine colors that define the morphotypes, represent only intraspecific variation of P. gaimardi . Eight polymorphic loci ( Cat , a -Est-1 , a -Est-2 , Mdh , Pep , Pgi , Pgm and Xod ) were assayed. Departures from Hardy-Weinberg equilibrium were tested at each locus in each morphotype from each locality. P. gaimardi species showed high levels of genetic variation (0.171-0.343) in accordance with the patterns observed for marine invertebrates. Diagnostic loci were not found among any of the five morphotypes and neither for the different geographic locations. A Mann Whitney U test showed values significantly lower in sympatry than in allopatry for both Nei’s Genetic Identities and 2 x 2 θ F-statistics. Furthermore, performance of AMOVA analysis considering a hierarchical levels of individuals, colour-morphs and locations indicates that the differences in genetic variation are sorted among collection sites rather than color morphotypes.

The genetic variability in selectively controlled in Bulgaria local honey bee populations, representing Apis mellifera macedonica subspecies (type “rodopica”), has been studied by usage of alloenzymic analysis of six enzymic systems (MDH-1, ME, EST-3, ALP, PGM and HK) corresponding to 6 loci. Totally 324 worker bee individuals from 9 different local populations belonging to breeding stock of National Bee Breeding Association were included in this investigation. All of the studied loci were found to be polymorphic in most of the populations with the exception of EST-3 locus which was established to be fixed in two of investigated populations. Polymorphism with three alleles was ascertained for MDH, ME, ALP and PGM loci and with four alleles – for EST-3 and HK loci. The most common alleles in all of the investigated populations were ME 100, EST-3 100, PGM 100 and HK 100. Two private alleles (frequency < 0.05) were found for two of the studied populations. The calculated level of polymorphism was between 88.33% and 100%. The observed and expected heterozygosities were found to range from 0.186 to 0.301, and from 0.205 to 0.305, respectively. The calculated mean Fst level was 0.028. Allele frequencies of all studied loci were used to estimate Nei’s (1972) genetic distance, which was established to range between 0.001 and 0.028 among the selectively controlled populations studied. The assignment test showed a high level of consolidation for the all studied populations.

In order to clarify the genetic differentiation and reproductive traits of mixed polyploid populations in Parasenecio kamtschaticus complex, geographical distribution, genetic diversity, and reproductive performance were compared among three intraspecific types composed of two cytotypes and two varieties in Japan. Diploid P. kamtschaticus var. kamtschaticus (2x-kamtschaticus) was distributed at the center of the distribution range, tetraploid P. kamtschaticus var. kamtschaticus (4x-kamtschaticus) existed widely throughout the entire range, and tetraploid P. kamtschaticus var. bulbifera (4x-bulbifera), producing bulbils, was restricted to higher elevations. Genetic structure was analyzed using allozyme markers. The genetic diversity of 4x-kamtschaticus was higher than that of 2x-kamtschaticus, with that of 4x-bulbifera being intermediate. Populations of 4x-bulbiferaand 2x-kamtschaticus were genetically discriminable from each other in principle coordinate analysis, and the genetic structure of 4x-kamtschaticus populations largely overlapped with those of the other types. Flower and achene production levels were highest in the 4x-kamtschaticus populations and lowest in the 4x-bulbifera populations. Germination activity of achenes was highest in the 2x-kamtschaticus populations and lowest in the 4x-bulbifera populations. Fruit-set success of 4x-bulbifera decreased with elevation because of a shorter growing season, indicating the importance of vegetative reproduction by bulbils at higher elevations. Unexpectedly, the inbreeding coefficients of the 4x-bulbifera populations were the lowest among the three types. Occasional achene production by outcrossing might maintain the high genetic diversity of the 4x-bulbifera populations. The evolution of polyploidy and subsequent bulbil production might enable P. kamtschaticus to disperse a wider range of environmental conditions.

Varijabilnost pčelinjih zajednica u Europi smanjuje se uslijed brojnih čimbenika. To je prvenstveno posljedica širenja raznih bolesti pčela (varooza, nozemoza i različite viroze), jednolične ishrane pčela (monokultura), upotrebe zaštitnih sredstava (pesticida) u biljnoj proizvodnji te trgovina i uvoz većinom dvije podvrste medonosne pčele, sive ili kranjske pčele (A. m. carnica) te žute ili talijanske pčele (A. m. ligustica). Iako su provedena brojna istraživanja, stvarni uzroci pada navedenog broja pčelinjih zajednica nisu u potpunosti jasni. S ciljem očuvanja lokalno prilagođenih podvrsta i ekotipova putem suvremenih uzgojnih programa, takve populacije je najprije potrebno utvrditi. U ovom radu bit će obrađene morfometrijske i molekularne metode koje su trenutno dostupne te se koriste za potrebe ispitivanja genetske specifičnosti i identifikaciju bioraznolikosti pčela, te bolji uvid u njihove taksonomske odnose., The natural variability of Western honey bee (Apis mellifera) in Europe is reduced due to a number of reasons. These are primarily consequences of spread of various bee diseases (varoosis, nosemosis, and variable viruses), uniform nutrition of bees (monoculture), significant use of pesticides in plant production and trade and import of mainly two subspecies (A. m. carnica and A. m. ligustica). The actual causes of the honey bee colony losses are not fully clear, although numerous studies have been carried out. In order to preserve locally adapted subspecies and ecotypes through contemporary breeding programs, such populations need to be deter- mined at first. In this article, the methods currently available, will be presented and used for investigation of genetic specificity and identification of bee biodiversity, also for better insight into their taxonomic relationships.

The Fagus sylvatica L. species complex in Europe and Western Asia comprises two commonly recognized subspecies, F. sylvatica subsp. sylvatica [= F. sylvatica sensu stricto (s. str.)] and F. sylvatica subsp. orientalis (= F. orientalis), and two putatively hybridogenous or intermediate taxa, “F. moesiaca” and “F. taurica”. The present study aimed to examine the demographic history of this species complex using 12 allelic loci of nine allozymes scored in 279 beech populations in western Eurasia. Three sets of phylogenetic scenarios were tested by approximate Bayesian computation: one dealing with the divergence of subspecies and/or regional populations within the whole taxonomical complex, and two others focusing on the potential hybrid origin of “F. moesiaca” and “F. taurica”. The best-supported scenario within the first set placed the time of divergence of regional populations of F. orientalis in the Early Pleistocene (1.18–1.87 My BP). According to this scenario, the Iranian population was the ancestral lineage, whereas F. sylvatica s. str. was the lineage that diverged most recently. “Fagus taurica” was found to have originated from hybridization between the Caucasian population of F. orientalis and F. sylvatica s. str. at 144 ky BP. In contrast, there was no evidence of a hybrid origin of “F. moesiaca”. The best-supported scenario suggested that the Balkan lineage is a part of F. sylvatica s. str., which diverged early from F. orientalis in Asia Minor (817 ky BP), while both the Italian and Central-European lineages diverged from the Balkan one later, at the beginning of the last (Weichselian) glacial period.

Australia was once thought to be a biodiversity desert when considering the subterranean world however, recent work has revealed a fascinating collection of cave creatures, many with surprising biogeographic histories. This has especially been so in the karstic regions of north-western Australia (Cape Range peninsula, Barrow Island, Pilbara), which is home not only to a diverse collection of subterranean invertebrates, but also to the continent's only known underworld-adapted vertebrates, which includes the cave fish in the genus Milyeringa. These cave gudgeons have recently been in a state of taxonomic flux, with species being both split and lumped, but this was done with limited data (incomplete geographic sampling and no nuclear DNA sequence data). Therefore, we have revisited the systematic status of Milyeringa in a total-evidence molecular approach by integrating all existing data (mitochondrial, allozymes) with new DNA sequences from nuclear and mitochondrial loci and new multilocus allozyme data. Our conclusion, that there are two species, matches the most recent taxonomic treatment, with Milyeringa veritas present on both the eastern and western sides of the Cape Range peninsula, and Milyeringa justitia on Barrow Island. This has implications for future research in the linked fields of biogeography and conservation Refereed/Peer-reviewed

Previous studies have shown that levels of genetic diversity in species of the genus Cephalanthera covary with the breeding system. In the southern part of the Korean Peninsula, the three self‐compatible terrestrial orchids Cephalanthera erecta, C. falcata and C. longibracteata flower synchronously in sympatric populations. The food‐deceptive C. falcata with bright yellow flowers is predominantly outcrossing, whereas autogamy is the dominant strategy in both C. erecta and C. longibracteata, whose white flowers do not open fully. We examined genetic diversity (by means of allozymes) of the three species in sympatric populations (600 × 600 m area) in the Yeonwhasan Provincial Park (YPP) and in non‐sympatric populations outside YPP, South Korea. Thirteen out of 20 putative loci were variable across the three species, but there was a complete lack of allozyme variation within each species and we found no evidence of hybridisation. Our results suggest that historical factors, i.e. the Quaternary climate oscillations, have played a major role in determining levels of genetic diversity in the three Cephalanthera species. The Korean populations of C. erecta (a warm‐temperate/temperate element) and C. falcata (a warm‐temperate element) may have been established by a single introduction from a genetically depauperate ancestral population, likely located outside the Korean Peninsula. On the other hand, since C. longibracteata is a boreal/temperate element, it may have survived the Last Glacial Maximum in microrefugia located in low elevation regions within the Peninsula where it has been subjected to population bottlenecks reducing its genetic diversity., This research was supported by a Korea Research Foundation grants; NRF‐2013R1A1A2063524 to MYC and NRF‐2013R1A1A3010892 and NRF‐2017R1A2B4012215 to MGC.

25 p., tablas, figura -- Postprint del artículo publicado en Botanical Review. Versión revisada y corregida, To provide basic information for orchid conservation, we surveyed the plant allozyme literature to summarize genetic diversity and structure data for (i) rare orchids native to the Korean Peninsula, and (ii) their congeners irrespective of being common and rare or Korean or not. A total of 68 taxa (32 taxa in Korea and 37 outside Korea; Goodyera repens being included in both datasets) were considered in this study. Overall, rare Korean orchid species had significantly lower levels of genetic diversity than their common congeners and common orchids in general at both population and species levels. However, mean values of GST (or FST) for rare and common orchids (Korean or not) did not differ significantly from each other. We found patterns of both low and high genetic diversity in rare Korean orchids. Many rare orchids harbored a complete lack of allozyme variation or extremely low within-population variation, perhaps due to rarity associated with random genetic drift and/or, for the case of warm-temperate orchids, to founder effects during post-glacial re-colonization. In contrast, high levels of genetic variation were found for a few orchids that have become recently rare (due to over-collection during the past several decades), probably because there have not been sufficient generations for the initial diversity to be substantially eroded. In addition, several orchids occurring in the main mountain system of the Korean Peninsula (the Baekdudaegan), that served as a glacial refugium, maintained moderate to high levels of within-population genetic diversity. Based on our genetic data, conservation priority should be given to rare orchid species. Particularly, urgent measures should be implemented on Jeju Island, a popular vacation spot, because it also a hotspot for threatened orchids with low levels of genetic diversity., This research was supported by Korea Research Foundation grants; KRF-2013R1A1A2063524 to M.Y. C. and NRF-2011-0017236, NRF-2013R1A1A3010892, and NRF-2017R1A2B4012215 to M. G. C. and was carried out as part of “Infrastructure for Conservation and Restoration of Rare and Endemic Plants in Korea National Arboretum” that supported to M. G. C. in 2015 to 2017.

The systematics of the genus Branchipus Schaeffer, 1766 (Branchiopoda: Anostraca) has been based on morphological taxonomy only and thus remains poorly resolved or, at best, questionable. Among the seven species described in the genus, a detailed comparison of Branchipus schaefferi Fischer, 1834 with both B. pasai Cottarelli, 1969 and B. visnyai Kertesz, 1956 is necessary to clarify their status. The use of morphological characters and, in particular, the appearance of second antennae and antennal appendages in males still cause taxonomic confusion among these three species. Herein, we use two molecular markers (allozymes and mtDNA sequences) and scanning electron microscopy to determine consistent differentiation patterns and test for congruence with the shape of the genus diagnostic trait, the frontal shield. We analyzed specimens belonging to 11 populations and covering a wide geographic distribution: B. pasai from Italy, B. schaefferi from Spain and Morocco, and B. visnyai from Malta and Italy. The extent of genetic divergence among populations evaluated by allozyme variation at 13 enzymatic loci and a mtDNA marker suggests the existence of different taxa and the absence of gene flow between them. The molecular results, together with morphological observations, did not provide support for the existing subdivision in the three nominal species B. pasai , B. schaefferi , and B. visnyai and places the validity of previously proposed taxonomic groups in discussion.

Data for electrophoretic pattern of 26 loci, resolved for the 8 protein systems (7- enzyme and 1-non-enzyme) assayed were used to investigate population structure of turbot along the Bulgarian and Romanian Black Sea coasts using genetic diversity measures. Seventeen loci were polymorphic in all populations and a total of 34 alleles were identified. Four types of tissue: muscle, retina, plasma and haemoglobin were analyzed. The percentage of polymorphic loci was high (65.38%) within populations. A low level of genetic differentiation among populations was detected, based on the Shannon’s information index (0.446-0.448) and the coefficient of genetic differentiation between populations (FST =0.014). The overall mean of within-population inbreeding estimate (FIS) was (-0.209) and demonstrated low level of inbreeding. The genetic distance (DNei) between the populations was low and vary between 0.003 and 0.014. Genetic distances among turbot populations were positively correlated with geographic distances (r = 0.474), but the association was not significant according to the Mantel test (p=0.651) and showed a lack of correlation between genetic distance and the geographic location of populations. Results identified one genetic stock with sufficient gene flow between all the three sites to prevent genetic differentiation from occurring. Only 1.4% of the genetic variation was observed among populations. Results revealed that adopting a single stock model and regional shared management could probably be appropriate for sustainable long-term use of turbot along western Black Sea coast. Determination of the contemporary state of the population distribution will be the prerequisite for determination of adequate measures for exploitation and protection of the existing turbot populations along western Black Sea coast.

Mosquitoes belonging to the Aedes mariae complex, including Aedes mariae (Sergent & Sergent), Aedes zammitii (Theobald), and Aedes phoeniciae (Coluzzi & Sabatini), are among the few animals inhabiting the extreme habitat of sea rock pools. Morphological analysis of these species and crossing experiments conducted in laboratory and natural conditions inferred the occurrence of three taxa with a parapatric distribution along the coasts of the Mediterranean Basin. To date, however, genetic characterization of the three species within the Ae. mariae complex and an assessment of their interspecific differentiation are lacking. In this study, we used both nuclear (i.e., 14 allozymic loci) and mitochondrial genetic markers (i.e., sequences of the cytochrome oxidase I and II genes) to genetically characterize the three species belonging to the complex. Then, we used mitochondrial markers to infer phylogenetic relationships between the species and estimate the time to most recent common ancestor (TMRCA). ...

Background. This article is about evolutionary processes in populations of small mobile species in the urbanized landscape. The evolutionary processes of a small mobile species in the urbanized landscape are considered in this article. Materials and methods. The morphological and genetic variety of the Helix pomatia L. populations was studied by the morphometric method and the polyacrylamyd gel electrophoresis method. Samples were collected from twelve landscape urbanized populations from south-eastern and eastern part of the current Helix pomatia L. area. Results. According to the data, the majority of the study populations of this mollusk have the satisfactory condition. The most populations are characterized as high level of heterozygosity, the low inbreeding and the large effective size values of the population testify to this. The structure of the gene pools in the populations snails in the study area is determined by the origin of these populations, the genetic-automatic processes and the microclimatic conditions of the urban environment.

Absorbing Hybridization of Cobitis taenia and Sabanejewia aurata (Cypriniformes, Cobitidae) in Water Reservoirs of Northern Ukraine Connected with Diploid-Polyploid Complex Formation. Mezhzherin, S. V., Pukhtayevych, P. P., Tsyba, А. А. — In the rivers of the Northern Ukraine by meams of karyological and allozyme analysys obtained extensive natural hybridization between Cobitis taenia and Sabanejewia aurata, initially leading to the formation of diploid hybrids. Besides that certain water bodies are predominantly populated by triploids of the C. taenia — C. tanaitica — S. aurata biotypes. Since the share of this kind of hybrid biotypes signifi cantly exceeds the number of individuals of S. aurata, it is concluded that the hybridization of a common and eurybiontic C. taenia with rarer stenobiontic S. aurata, is of absorbing character.

Hemiodus orthonops is a small fish of the Hemiodontidae family, order Characiformes, with a maximum of 25 cm standard length. Until recently, H. orthonops was an endemic species from the Paraná-Paraguay basin and it was absent from the upper Paraná River basin. Since 2008, it has started to be collected in the upper Paraná River, representing up to 10% of catches. Two population samples of H. orthonops from two localities of the upper Parana River basin (Porto Camargo and Porto Figueira) were analyzed using the allozymes electrophoresis technique. Twenty-one enzymatic loci were detected. The population sample from Porto Camargo displayed a genetic variability (He = 0.1061) higher than that from Porto Figueira (He = 0.0580) and homozygote excess in both of them. The FST value (0.2081) indicated genetic structure. The excess of homozygotes in both samples was probably due to founder effect in the population.

We investigated the evolutionary genetics of the marine bioinvader bivalve Isognomon bicolor (C. B. Adams, 1845) by the allozyme electrophoresis method. For that, we collected individuals in 2005, 2009, and 2013 at Praia de Itaipu (Niterói city, Rio de Janeiro state, southeastern Brazil). Seven allozymic systems were interpreted as eleven loci. The observed heterozygosity ranged from 0.324 in 2013 to 0.534 in 2005, but such differences were not statistically significant. High levels of genetic variation in bioinvaders can be explained as being a consequence of the Wahlund effect, which results in significant deviations from the Hardy-Weinberg equilibrium as well as in linkage disequilibrium. However, we observed no evidence of linkage disequilibrium, and only two loci showed significant deviations from the Hardy-Weinberg equilibrium (Mdh in 2005 and α-Est-1 in 2013). Temporal stability in the levels of genetic variation of I. bicolor at Praia de Itaipu may be an indication that the studied population has already reached the climax stage of the bioinvasion evolutionary process. Probably, in a first moment the Wahlund effect may have contributed for the population to respond to ecological pressures in a way contrary to population decline and extinction. In a second moment, the population may have reached maturity and climax, thus finally reaching the Hardy-Weinberg equilibrium for most loci., A genética evolutiva do bivalve marinho invasor Isognomon bicolor (C.B. Adams, 1845) foi investigada, pelo método de eletroforese de aloenzimas, para indivíduos coletados na Praia de Itaipu (Niterói – RJ) nos anos de 2005, 2009 e 2013. Sete sistemas enzimáticos foram interpretados como 11 loci gênicos. A heterozigosidade observada variou de 0,324 em 2013 a 0,534 em 2005, mas estas diferenças não foram significativas. Altos níveis de variação gênica em bioinvasores podem ser explicados como consequência do efeito Wahlund, que é acompanhado de desvios significativos ao equilíbrio Hardy-Weinberg e desequilíbrio de ligação entre loci. Não foram observados desequilíbrios de ligação e apenas dois loci apresentaram desvios significativos ao equilíbrio de Hardy-Weinberg (Mdh em 2005 e α-Est-1 em 2013). A estabilidade temporal dos níveis de variação gênica de I. bicolor na Praia de Itaipu pode ser uma indicação de que esta população tenha atingido a fase do processo evolutivo da bioinvasão conhecida como “clímax”. Num primeiro momento, o efeito Wahlund pode ter contribuído para que a população de Itaipu respondesse às pressões ecológicas no sentido contrário à redução populacional e à extinção. Em um segundo momento, esta população amadureceu para o “clímax” alcançando o equilíbrio de Hardy-Weinberg para maioria dos loci.

We investigated allozyme variation of the rare shrub Sophora koreensis in Yanggu County (South Korea), at the southern margin of the species range. To gain insights into ecological processes at the landscape level, we studied two or three populations from four localities (in total, 10 populations) in a range of ca. 6 km. We found low levels of within-population genetic variation (%P = 13.6, A = 1.14, and He = 0.026) and a moderate degree of among-population genetic differentiation (FST = 0.203). The analysis of molecular variance (AMOVA) revealed a substantially higher percentage of variation among populations within localities (17%) than among localities (5%). There was no significant relationship between pairwise genetic divergence and logarithm of pairwise geographic distance (r = 0.032, P = 0.842). These results suggest limited gene flow between populations within localities, while indicating that the optimal strategy for the preservation of the genetic variation of S. koreensis is to conserve as many populations as possible. Se investigó la variación isoenzimática del arbusto raro Sophora koreensis en el condado de Yanggu (Corea del Sur), en el extremo sur de su área de distribución. Para obtener información sobre los procesos ecológicos a nivel del paisaje, se estudiaron dos o tres poblaciones para cada una de las cuatro localidades (en total, 10 poblaciones) en un radio de unos 6 km. Se encontraron bajos niveles de variación genética intra-poblacional (%P = 13,6, A = 1,14 y He = 0,026) y un grado moderado de diferenciación genética entre poblaciones (FST = 0,203). El análisis de la varianza molecular (AMOVA) reveló un porcentaje sustancialmente mayor de variación entre poblaciones dentro cada una de las localidades (17%) que entre localidades (5%). No hubo una relación significativa entre la divergencia genética y el logaritmo de la distancia geográfica entre pares de poblaciones (r = 0,032, P = 0,842). Estos resultados sugieren un flujo genético limitado entre poblaciones dentro de las localidades, además de indicar que la mejor estrategia para la preservación de la diversidad genética de S. koreensis es la conservación del máximo número de poblaciones posible.

Fil: Meza Torres, Esteban I. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina. Fil: Stensvold, M. C. United States Department of Agriculture, Forest Service, Alaska Region; Estados Unidos. Fil: Stensvold, M. C. Iowa State University. Department of Ecology, Evolution and Organismal Biology; Estados Unidos. Fil: Farrar, D.R. Iowa State University. Department of Ecology, Evolution and Organismal Biology; Estados Unidos. Fil: Ferrucci, María Silvia. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina. Fil: Ferrucci, María Silvia. Consejo Nacional de Investigaciones Científicas y Técnica. Instituto de Botánica del Nordeste; Argentina. The genus Botrychium (Ophioglossaceae) is comprised of about 30 widely distributed species. Because of their small size and few, yet variable, morphological characters, many questionable taxa have been described over the years. Since its description in 1906, B. dusenii has been treated as both a distinct species and variously divided into four taxa: B. lunaria var. dusenii, B. lunaria var. typicum, B. lunaria var. antarctica, and B. ramosum var. patagonicum (B. matricariaefolium subsp. patagonicum). The aim of this study is to evaluate the validity of these taxa using spore wall sculpture, spore size, allelic composition employing enzyme electrophoresis, and morphological characters. We conclude that only one species of Botrychium occurs in southern South America and recognize this as B. dusenii, a species distinct from B. lunaria. Spore size and results of enzyme electrophoresis suggest that B. dusenii is an allotetraploid derived through hybridization between a diploid member of the B. lunaria complex and a diploid member of the B. campestre complex. We provide information about habitat, distribution, and full exsiccate citations.

Eight enzyme systems coding for 14 loci were assayed in Juniperus phoenicea. Mendelian inheritance was confirmed for allozymes at 10 loci (Fle 2, Gdh, Got 1, Idh, Mdh 1, Mdh 2, Men 1, Pgi, Pgm I, Pgm 2) by testing the fit of band-pattern segregation in macrogametophytes from heterozygous trees to the expected I: I ratio. Linkage relationships were examined for 21 pairs of allozyme loci and these pairs do not appear to be linked in our material.

Litvinchuk, S. N. et al., Based on allozyme variation of 410 newly collected individuals from 52 localities, we reconstructed range-wide phylogeography of the widespread Western Palearctic anuran, Pelobates fuscus. To study genetic diversity, evolutionary history, postglacial range expansion and secondary contact zones, we used a multidisciplinary approach combining information from various genetic analyses and ecological niche modeling. We confirmed the presence of two main groups in P. fuscus, initially revealed by genome size variation. Pelobates f. vespertinus presents a monomorphic group, but two main groups can be identified in P. f. fuscus: an East European and a West European group. We suggest the existence of at least four different Last Glacial refugia for P. fuscus: (1) the area between the Caspian and Azov Seas as the origin for the expansion of P. f. vespertinus; (2) the northwestern part of the Black Sea area for the East European P. f. fuscus; (3) the southwestern part of the Pannonian Plain and (4) the Po Plain for the West European P. f. fuscus. The routes of postglacial range expansions from the refugia are discussed. We newly identified a hybrid zone between P. f. fuscus and P. f. vespertinus. The width of this zone is about 12.5 km. In light of these findings, the two subspecies of P. fuscus constitute distinct evolutionary lineages and merit recognition as separate species. Our data do not provide support for the validity of P.f. insubricus. We therefore propose to synonymize this subspecies with P. f. fuscus. © 2013 Gesellschaft für Biologische Systematik., The work was partially funded by grants from the Russian Foundation of Basic Research (project nos. 12-04-01277 and 13-04-90410) and the Serbian Ministry of Education and Science and Technological Development (project no. 173043). PdP was funded by the FI program (Generalitat de Catalunya, Spain) and a grant from the Societas Europaea Herpetologica (2010). The work of AC was supported by a postdoctoral grant from Fundação para a Ciência e a Tecnologia (SFRH/BPD/72908/2010).

Paracentrotus gaimardi is a sea urchin reaching a maximum of 42 mm of diameter. It presents five morphotypes in sympatry characterized by their distinctive spine colors (e.g., green, black, gray, brown and rose). P. gaimardi is considered as a vulnerable species and it is included on the list of endangered species of the Brazilian coastline. In this work, allozyme electrophoresis was used to study the genetic variation among the five color morphotypes, from two different geographic locations along the coast of Rio de Janeiro State: Itaipu and Prainha, both in sympatry and allopatry. The underlying null hypothesis is that the different spine colors that define the morphotypes, represent only intraspecific variation of P. gaimardi. Eight polymorphic loci (Cat, a-Est-1, a-Est-2, Mdh, Pep, Pgi, Pgm and Xod) were assayed. Departures from Hardy-Weinberg equilibrium were tested at each locus in each morphotype from each locality. P. gaimardi species showed high levels of genetic variation (0.171-0.343) in accordance with the patterns observed for marine invertebrates. Diagnostic loci were not found among any of the five morphotypes and neither for the different geographic locations. A U-Mann Whitney test showed values significantly lower in sympatry than in allopatry for both Nei’s Geneti c Identities and 2 x 2 9 F-statistics. Furthermore, performance of AMOVA analysis considering hierarchical levels of individuals indicates that the differences in genetic variation are sorted among collection sites rather than color morphotypes. RESUMEN. Paracentrotus gaimardi es un pequeño erizo de mar que presenta un diámetro máximo de 42 mm. Este erizo presenta cinco morfotipos en simpatría caracterizados por sus distintivas espinas coloreadas (e.g., verde, negra, gris, café y rosa). P. gaimardi es considerada una especie vulnerable y está incluida en el listado de especies en peligro de la costa brasilera. En este trabajo se usaron caracteres aloenzimáticos para estudiar la variación genética entre cinco morfotipos de color, en dos localidades geográficas diferentes de la costa del Estado de Río de Janeiro: Itaipu y Prainha, ambas en simpatría y alopatría. La hipótesis nula analizada fue: si las diferentes espinas de color definen los morfotipos, representan solamente una variación intraespecífica de P. gaimardi. Ocho loci polimórficos (Cat, a-Est-1, a-Est-2, Mdh, Pep, Pgi, Pgm and Xod) fueron examinados. Las salidas del equilibrio de Hardy-Weinberg fueron examinadas en cada locus para cada morfotipo en cada localidad. P. gaimardi presenta grandes niveles de variación genética (0,171-0,343), lo cual concuerda con los parámetros observados para invertebrados marinos. No se encontró ningún loci diagnóstico en ninguno de los cinco morfotipos para ninguna de las áreas geográficas. La prueba del U-Mann Whitney mostró valores con baja significancia en simpatría y alopatría para las identidades genéticas de Nei y la prueba de 2x2 9 F. Adicionalmente, un análisis de AMOVA sugirió niveles jerárquicos para los individuos, indicando que las diferencias genéticas están organizadas alrededor de los sitios de colecta, más que en los morfotipos de color.

Allozymes were used to assess the genetic structure of 27 populations of Melampyrum subalpinum group and an artificial pollination experiment was carried out to examine the possibility of autogamy. Genetic variation was generally congruent with the known morphological variation of the group. The results corresponded with the central-marginal concept. Allelic enrichment due to hybridization with M. nemorosum was observed in some Austrian populations. Czech and Slovak populations do not differ from Austrian populations. The high inbreeding coefficient and the pollination experiment do not contradict the possibility of autogamy.

The genetic variability in selectively controlled in Bulgaria local honey bee populations, representing Apis mellifera macedonica subspecies (type “rodopica”), has been studied by usage of alloenzymic analysis of six enzymic systems (MDH-1, ME, EST-3, ALP, PGM and HK) corresponding to 6 loci. Totally 324 worker bee individuals from 9 different local populations belonging to breeding stock of National Bee Breeding Association were included in this investigation. All of the studied loci were found to be polymorphic in most of the populations with the exception of EST-3 locus which was established to be fixed in two of investigated populations. Polymorphism with three alleles was ascertained for MDH, ME, ALP and PGM loci and with four alleles – for EST-3 and HK loci. The most common alleles in all of the investigated populations were ME 100, EST-3 100, PGM 100 and HK 100. Two private alleles (frequency < 0.05) were found for two of the studied populations. The calculated level of polymorphism was between 88.33% and 100%. The observed and expected heterozygosities were found to range from 0.186 to 0.301, and from 0.205 to 0.305, respectively. The calculated mean Fst level was 0.028. Allele frequencies of all studied loci were used to estimate Nei’s (1972) genetic distance, which was established to range between 0.001 and 0.028 among the selectively controlled populations studied. The assignment test showed a high level of consolidation for the all studied populations., V nastoyashtoto izsledvane, na genetichen aloenzimen analiz sa podlozheni selektsionno kontrolirani populatsii medonosni pcheli ot devet (9) bazi na Natsionalnata Razvadna Asotsiatsiya po Pchelarstvo v Balgariya, prinadlezhashti kam podvid Apis mellifera macedonica (mesten tip “rodopica”). Ustanoveno e, che vsichkite izsledvani 6 aloenzimni lokusi sa polimorfni v pochti vsichki ot prouchvanite populatsii, s izklyuchenie na EST-3 lokusat, fiksiran s ediniya si alel (EST-3 100) v dve ot populatsiite. Trialelen polimorfizam e ustanoven za MDH, ME, ALP i PGM lokusite, a chetirialelen – za EST-3 i HK lokusi. Izsledvaneto sochi, che s naygolyama chestota na sreshtane v izsledvanite populatsii sa alelite ME 100, EST-3 100, PGM 100 i HK 100. Konstatirano e nalichieto na dva chastni alela (chestota < 0.05) v dve ot prouchvanite populatsii. Izchisleni sa niva na polimorfizam v diapazon ot 88.33% do 100%. Nablyudavanata i ochakvana heterozigotnost varirat mezhdu 0.186 i 0.301, i mezhdu 0.205 i 0.305, saotvetno. Izchislenoto sredno nivo na fiksatsionniya indeks (Fst) e 0.028. Ustanovenite genetichni distantsii po Nei (1972) sa v granitsata ot 0.001 do 0.028. Provedeniyat asignatsionen test (test za prinadlezhnost) pokazva visoko nivo na konsolidatsiya sred podlozhenite na selektsionen kontrol i vklyucheni v nastoyashtoto izsledvane populatsii.

Diversity among individuals in a population is an important feature linking vital rates with behaviour and spatial occupation. We measured the growth increments in the otolith of individual fishes collected on the annual fisheries survey PELGAS from 2001 to 2015. Individuals who grew larger at juvenile stage occupied later in life more off-shore habitats. Further, we analysed the allozymes of 13 different loci from 2001 to 2006. Alleles of the enzyme IDH showed different frequencies in inshore and offshore habitats. The population spatially segregates along a coast to off-shore gradient with individuals showing different early growth and allele frequencies. Results show how individuals in a population segregate spatially in different habitats in relation with phenotypic diversity. This implies modelling the population with individual-based and physiological approaches to fully grasp its dynamics. It also implies developing management strategies to conserve infra-population diversity as a means to garantee the occupation of the full range of habitats.

The burnet moth Zygaena anthyllidis, endemic to the high elevations of the Pyrenees, is vulnerable to land-use. In order to identify conservation priorities based on an assessment of genetic diversity within populations and gene flow among populations, we examined Z. anthyllidis' genetic variability and differentiation based on allozyme electrophoresis from seven populations scattered across its entire range. In comparison to other mountain Lepidoptera, the populations studied exhibit a low level of genetic diversity. Remarkable between-population differentiation ( F = 0.053), the presence of private alleles, and the lack of significant isolation-by-distance pattern characterises the genetic make-up of the species. We interpreted the pattern of genetic differentiation as a consequence of low dispersal power in combination with insufficient landscape connectivity. Ongoing land-use change might reinforce genetic differentiation due to habitat fragmentation and additionally affect negatively allozyme variability at shifting range margins, i.e. the capacity to adapt to changing environments. We therefore suggest creating a network of suitable habitats at the landscape scale to facilitate genetic exchange and to conserve the species' overall genetic variability. The burnet moth Zygaena anthyllidis, endemic to the high elevations of the Pyrenees, is vulnerable to land-use. In order to identify conservation priorities based on an assessment of genetic diversity within populations and gene flow among populations, we examined Z. anthyllidis' genetic variability and differentiation based on allozyme electrophoresis from seven populations scattered across its entire range. In comparison to other mountain Lepidoptera, the populations studied exhibit a low level of genetic diversity. Remarkable between-population differentiation (FST = 0. 053), the presence of private alleles, and the lack of significant isolation-by-distance pattern characterises the genetic make-up of the species. We interpreted the pattern of genetic differentiation as a consequence of low dispersal power in combination with insufficient landscape connectivity. Ongoing land-use change might reinforce genetic differentiation due to habitat fragmentation and additionally affect negatively allozyme variability at shifting range margins, i. e. the capacity to adapt to changing environments. We therefore suggest creating a network of suitable habitats at the landscape scale to facilitate genetic exchange and to conserve the species' overall genetic variability. © 2012 Springer Science+Business Media Dordrecht.

We examined the genetic structure and diversity of eight populations of Eryngium maritimum L. (Sea Holly) along 150 km coastline of the Gulf of Gdańsk in Poland. Allozyme electrophoresis revealed two polymorphic loci among the 31 we tested. The populations showed low levels of genetic diversity (P=6.25, A=1.062, Ht=0.026), with little genetic differentiation between populations (FST=0.03), although it was statistically significant (p

Allozyme genetic polymorphism in Bulgarian honey bee populations from four different locations in the south-eastern part of the Rhodopes Mountain was studied on six enzymic systems (MDH, ME, EST, ALP, PGM and HK) corresponding to six genetic loci. Allozyme analysis revealed that all studied loci were polymorphic in almost all investigated populations. The observed heterozygosity was found to range from 0.110 to 0.208 and Nei's genetic distance – between 0.016 and 0.061 among the studied populations. These honey bee populations were clustered in two groups in the UPGMA dendrogram. The Tihomir population was in a separate clade while other three populations (Kardzhali, Krumovgrad and Dolni Yurutci) were grouped together.

The phylogenetic relationships between naturally occurring Atlantic Crassostrea oyster species were inferred through analyses of mitochondrial (cytochrome oxidase subunit I and 16S) and nuclear (second internal transcribed spacer) sequences. We also scored 15 allozyme loci on 422 oys- ters to study population structuring of C. rhizophorae and C. brasiliana along 9000 km of the Western Atlantic coastline. Despite morphological similarities, C. virginica was genetically more closely related to C. rhizophorae than to C. brasiliana. In contrast, C. paraibanensis was genetically indistin- guishable from C. brasiliana, which is probably a junior synonym of the African C. gasar. Significant genetic differentiation between populations of C. rhizophorae and C. gasar were found along the Western Atlantic coast, supporting an isolation-by-distance pattern.

Tropical trees are generally long-lived making it difficult to assess the long-term effects of habitat fragmentation on genetic diversity. Maintenance of genetic diversity in fragmented landscapes is largely dependent on the species’ mating system and the degree of genetic connectivity (seed and pollen flow) among fragments. Currently, these parameters are largely unknown for many endangered tropical tree species. Additionally, landscape fragmentation may isolate tropical tree populations from larger, more continuous populations. The role of isolated individuals in pollen transfer within and between remnant populations is not clear. In this study, we estimate the mating system and pollen flow patterns in continuous and remnant populations of the endangered tropical tree Guaiacum sanctum (Zygophyllaceae). Fractional paternity analyses were used to estimate average gene flow distances between fragmented remnant populations and the siring success of an intermediately located, but isolated individual. In these populations, G. sanctum is a mixed-mating species (t m = 0.72 − 0.95) whose pollen is transported over large distances (>4 km). An isolated tree may have functioned as a stepping-stone between two clusters of individuals, assisting long-distance pollen movement. This individual also sired a disproportionately high number of seeds (13.9%), and is thus an important component of the reproductive success of these populations, thus rejecting Janzen’s “living-dead” hypothesis. The high levels of genetic diversity maintained as a consequence of long-distance pollen-flow suggest that this endangered species may have the potential for future adaptation and population expansion if suitable habitats become available. Universidad de Costa Rica Organization for Tropical Studies National Science Foundation UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Biología