Jerzy Sell

Host compatibility as a critical factor in management unit recognition: population‐level differences in mussel–fish relationships

Summary 1. The recognition of management units (MUs) that respect interpopulation distinctions in management needs is central to many biological applications addressing species conservation, biological invasions and ecosystem processes. 2. We present a methodological approach for the evaluation of population-level differences in the host compatibility of natural populations of affiliate (dependent) species. Two experiments were performed to diagnose the sources of variability in the relationships between an endangered freshwater mussel Unio crassus and its host fish species in a fragmented river system in Central Europe. 3. Despite the common approach applied to the conservation of U. crassus throughout the study area, we identified differences in the ability of U. crassus to infest particular host fish species between nearby and recently isolated mussel populations. These differences could strongly influence their reproductive success under a constant conservation regime. 4. The observed differences were accompanied by genetic and morphometric distinctions, as revealed by microsatellite, mtDNA and elliptic Fourier descriptor analyses. The genetic data indicated substantial reproductive isolation, which could have resulted in adaptive or random changes responsible for the differences in host compatibility. 5. We showed that experimental testing of physiological host compatibility can be effectively used for the detection of different MUs of U. crassus, with direct implications for the management of host populations. In combination with traditional genetic and morphometric techniques, this approach may provide a general framework for the recognition and care of management units of affiliate species. 6. Synthesis and applications. Population-level evaluations of host compatibility should be utilized in management unit recognition and the formulation of management targets for affiliate species. We demonstrated that small-scale cross-compatibility testing can be effectively used to diagnose the sources of variability in host relationships with direct management implications. This approach can enhance the targeting of management actions in many biological applications addressing species conservation, biological invasions and ecosystem processes.

A new view on dam lines in Polish Arabian horses based on mtDNA analysis

Polish Arabian horses are one of the oldest and the most important Arab populations in the world. The Polish Arabian Stud Book and the Genealogical Charts by Skorkowski are the main sources of information on the ancestors of Polish Arabs. Both publications were viewed as credible sources of information until the 1990s when the data regarding one of the dam lines was questioned. The aim of the current study was to check the accuracy of the pedigree data of Polish dam lines using mtDNA analysis. The analyses of a 458 bp mtDNA D-loop fragment from representatives of 15 Polish Arabian dam lines revealed 14 distinct haplotypes. The results were inconsistent with pedigree data in the case of two lines. A detailed analysis of the historical sources was performed to explain these discrepancies. Our study revealed that representatives of different lines shared the same haplotypes. We also noted a genetic identity between some lines founded by Polish mares of unknown origin and lines established by desert-bred mares.

Identification and Characterization of the First Microsatellite Loci for the Thick-Shelled River Mussel Unio crassus (Bivalvia: Unionidae)

The thick-shelled river mussel Unio crassus is critically endangered throughout its range as a result of increasing human activity and habitat loss. Next generation DNA sequencing was used to develop a set of microsatellite markers that can be used for future ecological and population genetics studies of U. crassus. A total of 11 polymorphic loci were identified and characterized using 57 individuals from two Polish populations. Numbers of alleles ranged from 2 to 15 and expected heterozygosity levels ranged from 0.069 to 0.899. Deficiency of heterozygous genotypes was observed in four loci. Marker independence was confirmed with tests for linkage disequilibrium, however, analyses indicated evidence of null alleles in four loci. The microsatellite markers developed specifically for U. crassus provide a valuable tool for future ecological, population genetic assessments, and conservation management of this species.

Low genetic diversity in Polish populations of sibling ant species: Lasius niger (L.) and Lasius platythorax Seifert (Hymenoptera, Formicidae)

We present preliminary data on mitochondrial DNA diversity within and among populations of the ants Lasius niger and Lasius platythorax in Poland. Phylogenetic analysis based on the mitochondrial DNA markers: cytochrome c oxidase subunit I (cox1) and 16S ribosomal RNA (16S rRNA) confirms the species status of L. niger and L. platythorax. Intraspecific variability is low in both species, which might be a result of severe bottlenecks and rapid postglacial expansion into Central Europe.

Gender-Associated Mitochondrial DNA Heteroplasmy in Somatic Tissues of the Endangered Freshwater Mussel Unio crassus (Bivalvia: Unionidae): Implications for Sex Identification and Phylogeographical Studies.

Some bivalve species possess two independent mitochondrial DNA lineages: maternally (F-type) and paternally (M-type) inherited. This phenomenon is called doubly uniparental inheritance. It is generally agreed that F-type mtDNA is typically present in female somatic and gonadal tissues as well as in male somatic tissues, whereas the M-type mtDNA occurs only in male germ line and gonadal tissue. In the present study, the mtDNA heteroplasmy (for both F and M genomes) in male somatic tissues of Unio crassus (Philipsson, 1788), species threatened with extinction, has been confirmed. Taking advantage from the presence of Mcox1 marker only in male somatic tissues, we developed a new method of sex identification in this endangered species, using nondestructive tissue sampling. Probability of correct sex identification was estimated at 97.5%. The present study is the first report on gender-associated mitochondrial DNA heteroplasmy in male somatic tissues of thick-shelled river mussel and first approach to U. crassus sex identification at molecular level. Our study also confirmed the utility of paternally inherited Mcox1 gene fragment as a complementary molecular tool for resolving phylogeographical relationships among populations of thick-shelled river mussel.

The complete maternal and paternal mitochondrial genomes of Unio crassus: Mitochondrial molecular clock and the overconfidence of molecular dating

The availability of a rapidly growing number of complete mitochondrial genome sequences provokes high confidence dating approaches. However, even if the congruence between mitochondrial and nuclear markers is reasonable, the resulting topologies are frequently questionable. The unique opportunity to study the evolutionary history of two independent mitochondrial genomes in one phylogenetic context exists in the freshwater mussels family Unionidae. The two lineages function under doubly uniparental inheritance since well before the emergence of the family. Despite the relatively high number of available complete sequences of maternally inherited genomes, comparative analyses are limited by the small number of sequences of counterpart paternally inherited genomes. We have sequenced for the first time the representative set of five sequences (two maternal and three paternal) from the species Unio crassus. Comparative analysis of the phylogenies reconstructed using relevant mitogenomic data available in GenBank (13 species in total) reveal that single - genome inferences are congruent only if the relaxed clock is assumed.

Patterns of genetic differentiation and population history of endemic isopods (Asellidae) from ancient Lake Ohrid: combining allozyme and mtDNA data

Ancient lakes as places of extensive speciation processes have been characterized by a high degree of endemicity and biodiversity. The most outstanding European ancient lake is the oligotrophic and karstic Balkan Lake Ohrid. The lake is inhabited by a number of endemic species, but their evolutionary history is largely unresolved. in the present study, the genetic structure, gene genealogy and demographic history of the representatives of the Ohridian endemic Proasellus species were studied using both biparentally (allozyme loci) and maternally (partial mitochondrial cytochrome oxidase subunit I gene) inherited markers. Both data sets gave similar results and supported discrepancies among genetic differentiation, the current morphology-based taxonomy and bathymetric segregation. Horizontal distribution of endemic Proasellus species (Lake Ohrid vs adjacent feeder springs) within the lake presumably promote parapatric speciation whereas the main role of vertical barriers into diversification processes was not fully supported. The analyses of demographic history suggested the decline of endemic isopod populations. The radiation of endemic Proasellus populations within the lake could have started from the sublittoral/profundal zone towards the littoral or in the opposite direction — from the littoral to the profundal. Our analyses did not exclude both possibilities.

Morphologically Intermediate form between Athous haemorrhoidalis and A. vittatus (Coleoptera: Elateridae): A Case of Hybridization? a Preliminary Study

Abstract. In the present preliminary study we report an investigation on taxonomic status of individuals displaying diagnostic traits of intermediate values between the morphologically typical forms of Athous haemorrhoidalis (Fabricius 1801) and A. vittatus (Gmelin 1790) species occurring sympatrically. The presence of the “intermediate form” invoked the hypothesis of interspecific hybridization between A. haemorrhoidalis and A. vittatus species. An alternative explanation is that the “intermediate form” comprises individuals that are morphological variants of either of the species, without interspecific gene flow. We used partial sequences of two mitochondrial genes coding for the cytochrome oxidase subunit I (cox1) and 16S ribosomal RNA (rrnL), respectively. Additionally, we examined the variability of five polymorphic allozyme loci: 6Pgd, Idh, Mdh, Pgm and Gpi. Altogether, 28 A. haemorrhoidalis, 7 A. vittatus and 8 “intermediate form” individuals were included into the study. Phylogenetic analyses produced the trees of a consistent topology with high statistical support regardless of the method used. The specimens signified as the “intermediate form” constituted a monophyletic group with individuals of A. vittatus species. Empirical data confirmed that the reproductive isolation between the species studied is complete. No haplotype was shared between A. haemorrhoidalis and A. vittatus in sympatry. Similar results were obtained using the analysis of allozyme data. Especially at Idh and 6Pgd loci, alleles characteristic for A. vittatus and the “intermediate form” were not found in A. haemorrhoidalis, despite the larger sample examined. This suggests that the “intermediate form” is a phenotypic variant of A. vittatus without the signature of introgression.

Genetic Variability in Natural Populations of Eurytopic Ostracod Candona neglecta Sars

Abstract An electrophoretic survey of allozyme variation was conducted in four, highly polymorphic loci on nine populations of ostracod Candona neglecta Sars from three different environments: the profundal of post-glacial lakes, deep muddy bottom of the Baltic Sea and small astatic water bodies. The results suggest lack of genetic isolation between populations from lake profundal and the Baltic Sea. On the other hand a very distinct founder effect can be noted in the case of young, isolated populations from small astatic basins. It is suggested that a population inhabiting a large lake may be genetically subdivided due to differentiated eutrophication.

Instances of erroneous DNA barcoding of metazoan invertebrates: Are universal cox1 gene primers too “universal”?

The cytochrome c oxidase subunit I (cox1) gene is the main mitochondrial molecular marker playing a pivotal role in phylogenetic research and is a crucial barcode sequence. Folmer’s “universal” primers designed to amplify this gene in metazoan invertebrates allowed quick and easy barcode and phylogenetic analysis. On the other hand, the increase in the number of studies on barcoding leads to more frequent publishing of incorrect sequences, due to amplification of non-target taxa, and insufficient analysis of the obtained sequences. Consequently, some sequences deposited in genetic databases are incorrectly described as obtained from invertebrates, while being in fact bacterial sequences. In our study, in which we used Folmer’s primers to amplify COI sequences of the crustacean fairy shrimp Branchipus schaefferi (Fischer 1834), we also obtained COI sequences of microbial contaminants from Aeromonas sp. However, when we searched the GenBank database for sequences closely matching these contaminations we found entries described as representatives of Gastrotricha and Mollusca. When these entries were compared with other sequences bearing the same names in the database, the genetic distance between the incorrect and correct sequences amplified from the same species was c.a. 65%. Although the responsibility for the correct molecular identification of species rests on researchers, the errors found in already published sequences data have not been re-evaluated so far. On the basis of the standard sampling technique we have estimated with 95% probability that the chances of finding incorrectly described metazoan sequences in the GenBank depend on the systematic group, and variety from less than 1% (Mollusca and Arthropoda) up to 6.9% (Gastrotricha). Consequently, the increasing popularity of DNA barcoding and metabarcoding analysis may lead to overestimation of species diversity. Finally, the study also discusses the sources of the problems with amplification of non-target sequences.