Aphids and Ants, Mutualistic Species, Share a Mariner Element with an Unusual Location on Aphid Chromosomes

Aphids (Hemiptera, Aphididae) are small phytophagous insects. The aim of this study was to determine if the mariner elements found in the ant genomes are also present in Aphis fabae and Aphis hederae genomes and the possible existence of horizontal transfer events. Aphids maintain a relationship of mutualism with the ants. The close contact between these insects could favour horizontal transfer events of transposable elements. Myrmar mariner element isolated from Myrmica ruginodis and Tapinoma ibericum ants have also been found in the two Aphis species: A. fabae and A. hederae (Afabmar-Mr and Ahedmar-Mr elements). Besides, Afabmar-Mr could be an active transposon. Myrmar-like elements are also present in other insect species as well as in one Crustacean species. The phylogenetic study carried out with all Myrmar-like elements suggests the existence of horizontal transfer. Most aphids have 2n = 8 with a XX-X0 sex determination system. Their complicated life cycle is mostly parthenogenetic with sexual individuals only in autumn. The production of X0 males, originated by XX females which produce only spermatozoa with one X chromosome, must necessarily occur through specialized cytogenetic and molecular mechanisms which are not entirely known. In both aphid species, the mariner elements are located on all chromosomes, including the X chromosomes. However, on the two X chromosomes, no positive signals are detected in their small DAPI-negative telomere regions. The rDNA sites are located, as in the majority of Aphids species, on one of the telomere regions of each X chromosome. The hybridization patterns obtained by double FISH demonstrate that Afabmar-Mr and Ahedmar-Mr elements do not hybridize at the rDNA sites of their host species. Possible causes for the absence of these transposons in the rDNA genes are discussed, probably related with the X chromosome biology.

Evolutionary Tracks of Chromosomal Diversification in Surgeonfishes (Acanthuridae: Acanthurus) Along the World’s Biogeographic Domains

Fishes of the genus Acanthurus (Acanthuridae) are strongly related to reef environments, in a broad biogeographic context worldwide. Although their biological aspects are well known, cytogenetic information related to this genus remains incipient. In this study, Acanthurus species from populations inhabiting coastal regions of the Southwest Atlantic (SWA), South Atlantic oceanic islands (Fernando de Noronha Archipelago and Trindade Island), Greater Caribbean (GC), and Indo-Pacific Ocean (the center of the origin of the group) were analyzed to investigate their evolutionary differentiation. For this purpose, we employed conventional cytogenetic procedures and fluorescence in situ hybridization of 18S rDNA, 5S rDNA, and H3 and H2B-H2A histone sequences. The Atlantic species (A. coeruleus, A. chirurgus, and A. bahianus) did not show variations among them, despite their vast continental and insular distribution. In contrast, A. coeruleus from SWA and GC diverged from each other in the number of 18S rDNA sites, a condition likely associated with the barrier created by the outflows of the Amazonas/Orinoco rivers. The geminate species A. tractus had a cytogenetic profile similar to that of A. bahianus. However, the chromosomal macrostructures and the distribution of rDNA and hisDNA sequences revealed moderate to higher rates of diversification when Acanthurus species from recently colonized areas (Atlantic Ocean) were compared to A. triostegus, a representative species from the Indian Ocean. Our cytogenetic data covered all Acanthurus species from the Western Atlantic, tracked phylogenetic diversification throughout the dispersive process of the genus, and highlighted the probable diversifying role of ocean barriers in this process.

Analyses of the Updated “Animal rDNA Loci Database” with an Emphasis on Its New Features

We report on a major update to the animal rDNA loci database, which now contains cytogenetic information for 45S and 5S rDNA loci in more than 2600 and 1000 species, respectively. The data analyses show the following: (i) A high variability in 5S and 45S loci numbers, with both showing 50-fold or higher variability. However, karyotypes with an extremely high number of loci were rare, and medians generally converged to two 5S sites and two 45S rDNA sites per diploid genome. No relationship was observed between the number of 5S and 45S loci. (ii) The position of 45S rDNA on sex chromosomes was relatively frequent in some groups, particularly in arthropods (14% of karyotypes). Furthermore, 45S rDNA was almost exclusively located in microchromosomes when these were present (in birds and reptiles). (iii) The proportion of active NORs (positively stained with silver staining methods) progressively decreased with an increasing number of 45S rDNA loci, and karyotypes with more than 12 loci showed, on average, less than 40% of active loci. In conclusion, the updated version of the database provides some new insights into the organization of rRNA genes in chromosomes. We expect that its updated content will be useful for taxonomists, comparative cytogeneticists, and evolutionary biologists.

Tracking the Evolutionary Trends Among Small-Size Fishes of the Genus Pyrrhulina (Characiforme, Lebiasinidae): New Insights From a Molecular Cytogenetic Perspective

Miniature fishes have always been a challenge for cytogenetic studies due to the difficulty in obtaining chromosomal preparations, making them virtually unexplored. An example of this scenario relies on members of the family Lebiasinidae which include miniature to medium-sized, poorly known species, until very recently. The present study is part of undergoing major cytogenetic advances seeking to elucidate the evolutionary history of lebiasinids. Aiming to examine the karyotype diversification more deeply in Pyrrhulina, here we combined classical and molecular cytogenetic analyses, including Giemsa staining, C-banding, repetitive DNA mapping, comparative genomic hybridization (CGH), and whole chromosome painting (WCP) to perform the first analyses in five Pyrrhulina species (Pyrrhulina aff. marilynae, Pyrrhulina sp., P. obermulleri, P. marilynae and Pyrrhulina cf. laeta). The diploid number (2n) ranged from 40 to 42 chromosomes among all analyzed species, but P. marilynae is strikingly differentiated by having 2n = 32 chromosomes and a karyotype composed of large meta/submetacentric chromosomes, whose plesiomorphic status is discussed. The distribution of microsatellites does not markedly differ among species, but the number and position of the rDNA sites underwent significant changes among them. Interspecific comparative genome hybridization (CGH) found a moderate divergence in the repetitive DNA content among the species’ genomes. Noteworthy, the WCP reinforced our previous hypothesis on the origin of the X1X2Y multiple sex chromosome system in P. semifasciata. In summary, our data suggest that the karyotype differentiation in Pyrrhulina has been driven by major structural rearrangements, accompanied by high dynamics of repetitive DNAs.

Adding New Pieces to the Puzzle of Karyotype Evolution in Harttia (Siluriformes, Loricariidae): Investigation of Amazonian Species

A remarkable morphological diversity and karyotype variability can be observed in the Neotropical armored catfish genus Harttia. These fishes offer a useful model to explore both the evolution of karyotypes and sex chromosomes, since many species possess male-heterogametic sex chromosome systems and a high rate of karyotype repatterning. Based on the karyotype organization, the chromosomal distribution of several repetitive DNA classes, and the rough estimates of genomic divergences at the intraspecific and interspecific levels via Comparative Genomic Hybridization, we identified shared diploid chromosome numbers (2n = 54) but different karyotype compositions in H. dissidens (20m + 26sm + 8a) and Harttia sp. 3 (16m + 18sm + 14st + 6a), and different 2n in H. guianensis (2n = 58; 20m + 26sm + 2st + 10a). All species further displayed similar patterns of chromosomal distribution concerning constitutive heterochromatin, 18S ribosomal DNA (rDNA) sites, and most of the surveyed microsatellite motifs. Furthermore, differences in the distribution of 5S rDNA sites and a subset of microsatellite sequences were identified. Heteromorphic sex chromosomes were lacking in H. dissidens and H. guianensis at the scale of our analysis. However, one single chromosome pair in Harttia sp. 3 males presented a remarkable accumulation of male genome-derived probe after CGH, pointing to a tentative region of early sex chromosome differentiation. Thus, our data support already previously outlined evidence that Harttia is a vital model for the investigation of teleost karyotype and sex chromosome dynamics.

Spontaneous generation of diversity in Arachis neopolyploids (A. ipaënsis x A. duranensis)4x replays the early stages of peanut evolution

Polyploidy is considered a driving force in plant evolution and domestication. Although in the genus Arachis, several diploid species were traditionally cultivated for their seeds, only the allotetraploid peanut Arachis hypogaea became the successful, widely spread legume crop. This suggests that polyploidy has given selective advantage for domestication of peanut. Here we study induced allotetraploid (neopolyploid) lineages obtained from crosses between the peanut’s progenitor species, A. ipaënsis and A. duranensis, at earlier and later generations. We observed plant morphology, seed dimensions, and genome structure using cytogenetics (FISH and GISH) and SNP genotyping. The neopolyploid lineages show more variable fertility and seed morphology than their progenitors and cultivated peanut. They also showed sexual and somatic genome instability, evidenced by changes of number of detectable 45S rDNA sites, and extensive homoeologous recombination indicated by mosaic patterns of chromosomes and changes in dosage of SNP alleles derived from the diploid species. Genome instability was not randomly distributed across the genome: the more syntenic chromosomes, the higher homoeologous recombination. Instability levels are higher than observed on peanut lines, therefore it is likely that more unstable lines tend to perish. We conclude that early stages of the origin and domestication of the allotetraploid peanut involved two genetic bottlenecks: the first, common to most allotetraploids, is composed of the rare hybridization and polyploidization events, followed by sexual reproductive isolation from its wild diploid relatives. Here we suggest a second bottleneck: the survival of the only very few lineages that had stronger mechanisms for limiting genomic instability.

First cytogenetic characterization of the Amazon Catfish Leiarius marmoratus (Gill, 1870) and its hybrid with Pseudoplatystoma reticulatum (Eigenmann & Eigenmann, 1889)

This study reports the first cytogenetic characterization of the Amazonian catfish Leiarius marmoratus (“jandiá”) and its F1 (first generation) hybrid “cachandiá” with Pseudoplatystoma reticulatum (“cachara”). A diploid number of 56 chromosomes and a single argyrophilic nucleolus organizer region (Ag-NOR) in the short arm of two sub-telocentric chromosomes were observed for both L. marmoratus and P. reticulatum, but with differences in the karyotype formula and the size of the chromosome pair with NORs. The hybrid showed 2n = 56 chromosomes with an intermediate karyotype when compared to the parental species. A single Ag-NOR was maintained in the hybrid but located in two chromosomes with marked differences in size and presenting intraindividual variation in NOR activity (nucleolar dominance). For L. marmoratus and the hybrid, heterochromatic bands were predominately distributed in the terminal, centromeric, and sub-centromeric regions of some chromosomes and 5S rDNA sites located in two distinct sub-telocentric chromosomes, similar to the previously described for P. reticulatum. The data suggested that the hybrid karyotype might be insufficient for a precise discrimination of hybrids, however, Ag-NOR can be used as a chromosome marker to differentiate “cachandiá” from L. marmoratus and P. reticulatum. The current study also provides insights into the chromosomal features of L. marmoratus and contributes with novel cytogenetic information of this native Amazonian catfish included in the Pimelodidae family.

Composition and Nature of Heterochromatin in the Electrical Fish (Knifefishes) Gymnotus (Gymnotiformes: Gymnotidae)

Fishes of the genus Gymnotus have been suggested as a good model for biogeographic studies in the South American continent. In relation to heterochromatin, species of this genus have blocks preferably distributed in the centromeric region. The content of these regions has been shown to be variable, with description of transposable elements, pseudogenes of 5S rDNA and satellite sequences. In G. carapo Clade, although geographically separated, species with 2n = 54 chromosomes share the distribution of many 5S rDNA sites, a unique case within the genus. Here, repetitive DNA sequences from G. sylvius (2n = 40) and G. paraguensis (2n = 54) were isolated and mapped to understand their constitution. The chromosome mapping by FISH showed an exclusive association in the centromeres of all chromosomes. However, the cross-FISH did not show positive signs of interspecific hybridization, indicating high levels of heterochromatic sequence specificity. In addition, COI-1 sequences were analyzed in some species of Gymnotus, which revealed a close relationship between species of clade 2n = 54, which have multiple 5S rDNA sites. Possibly, the insertion of retroelements or pseudogenization and dispersion of this sequence occurred before the geographic dispersion of the ancestor of this clade from the Amazon region to the hydrographic systems of Paraná-Paraguay, a synapomorphy for the group.

Cytogenetic Analysis, Heterochromatin Characterization and Location of the rDNA Genes of Hycleus scutellatus (Coleoptera, Meloidae); A Species with an Unexpected High Number of rDNA Clusters

Meloidae are commonly known as blister beetles, so called for the secretion of cantharidin, a toxic substance that causes irritation and blistering. There has been a recent increase in the interest of the cantharidin anticancer potential of this insect group. Cytogenetic and molecular data in this group are scarce. In this study, we performed a karyotype analysis of Hycleus scutellatus, an endemic species of the Iberian Peninsula. We determined its chromosome number, 2n = 20, as well as the presence of the X and Y sex chromosomes. In addition to a karyotype analysis, we carried out DAPI staining. By fluorescence in situ hybridization we mapped the rDNA clusters on 12 different chromosomes. Compared to others, this species shows an unusually high number of chromosomes carrying rDNA. This is one of the highest numbers of rDNA sites found in the Polyphaga suborder (Coleoptera). Additionally, we isolated a satellite DNA family (Hyscu-H), which was located within the pericentromeric regions of all chromosomes, including the sex chromosomes. The results suggest that Hyscu-H is likely to be one of the most abundant satellite DNA repeats in H. scutellatus.

Origin and evolution of highly polymorphic rDNA sites in Alstroemeria longistaminea (Alstroemeriaceae) and related species

<i>Alstroemeria</i> (Alstroemeriaceae) displays a conserved and highly asymmetric karyotype, where most rDNA sites can be properly recognized by the size and morphology of the chromosomes. We analyzed the intraspecific variation of rDNA sites in <i>A. longistaminea </i>and compared with their distribution in other species (<i>A. caryophyllaea</i> and <i>A. piauhyensis</i>) and a representative of the sister genus, Bomarea edulis. All three <i>Alstroemeria</i> species presented 2n = 16 and one to six B chromosomes were found in some individuals of <i>A. longistaminea</i>. There was a set of 12 conserved rDNA sites (four 5S and eight 35S) and up to 11 variable sites. B chromosomes were almost entirely covered by 35S signals, coupled with tiny 5S sites. Noteworthy, most rDNA sites found in <i>A. caryophyllaea</i> and <i>A. piauhyensis</i> were localized in chromosome positions similar to those in <i>A. longistaminea</i>, suggesting the existence of conserved hotspots for rDNA accumulation. Some of these hotspots were absent in Chilean <i>Alstromeria</i> as well in <i>B. edulis</i>. We propose that insertions of rDNA sequences on chromosomes do not occur randomly but rather on preferential sites or hotspots for insertions. The maintenance of these arrays, however, may be favored/constrained by different factors, resulting in stable or polymorphic sites.