Cytogenetic analysis of two related Deltochilum (Coleoptera, Scarabaeidae) species: Diploid number reduction, extensive heterochromatin addition and differentiation

Departamento de Genética, Centro de Ciências Biológicas/CCB, Universidade Federal de Pernambuco/UFPE, Recife, Pernambuco, Brazil.
Micron (Impact Factor: 1.99). 10/2009; 41(2):112-7. DOI: 10.1016/j.micron.2009.10.005
Source: PubMed


Male mitotic and meiotic chromosomes of two species of the genus Deltochilum (Scarabaeidae) were analyzed through conventional staining, C-banding, base-specific fluorochromes, silver nitrate staining (AgNO(3)) and FISH (45S rDNA). The two species possessed karyotypes with 2n=14, neo-XY and meta-submetacentric chromosomes. The analysis of constitutive heterochromatin (CH) revealed mainly diphasic chromosomes in the two species, showing heterochromatic long arms. Silver nitrate staining labeled the blocks corresponding to CH in D. (Deltohyboma) aff morbillosum while in D. (Deltohyboma) calcaratum, AgNO(3) staining revealed only the CH blocks of the diphasic autosomes. The fluorochrome staining revealed in D. (D.) calcaratum the diphasic autosomes and the sex chromosomes with CMA(3)(+) blocks, and in D. (D.) aff morbillosum, the GC-rich sequences were restricted to the terminal regions of the long arms of the pairs 1 and 2 and the X. The FISH revealed 45S rDNA sites in two autosomic pairs and in the X chromosome. The analyses performed allowed for the identification of cytogenetic markers and the discussion of possible chromosome rearrangements that have been involved in the karyotypic differentiation of these species mainly related to the repetitive genome.

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    • "The PCR products containing a pool of Mariner sequences, and the C 0 t-1 DNA fraction were labeled with biotin-11-dATP by nick translation using the Bionick Labeling System kit (Invitrogen, San Diego, CA, USA). The FISH protocol followed the adaptations described by Cabral-de-Mello et al. [79]. The probes were labeled with biotin-14-dATP and detected by avidin-FITC (fluorescein isothiocynate) conjugated (Sigma-Aldrich, St. Louis, MO, USA). "
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    ABSTRACT: Background With the aim to increase the knowledge on the evolution of coleopteran genomes, we investigated through cytogenetics and nucleotide sequence analysis Mariner transposons in three Scarabaeinae species (Coprophanaeus cyanescens, C. ensifer and Diabroctis mimas). Results The cytogenetic mapping revealed an accumulation of Mariner transposon in the pericentromeric repetitive regions characterized as rich in heterochromatin and C0t-1 DNA fraction (DNA enriched with high and moderately repeated sequences). Nucleotide sequence analysis of Mariner revealed the presence of two major groups of Mariner copies in the three investigated coleoptera species. Conclusions The Mariner is accumulated in the centromeric area of the coleopteran chromosomes probably as a consequence of the absence of recombination in the heterochromatic regions. Our analysis detected high diversification of Mariner sequences during the evolutionary history of the group. Furthermore, comparisons between the coleopterans sequences with other insects and mammals, suggest that the horizontal transfer (HT) could have acted in the spreading of the Mariner in diverse non-related animal groups.
    Molecular Cytogenetics 11/2013; 6(1). DOI:10.1186/1755-8166-6-54 · 2.14 Impact Factor
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    • "The great amount of constitutive heterochromatin (CH) observed in the karyotypes of the species analyzed here was also described in other Scarabaeinae representatives , such as Deltochilum calcaratum, D. aff. morbillosum (Deltochilini) and some Phanaeini species, such as Coprophanaeus (Coprophanaeus) cyanescens, C. (Megaphanaeus) ensifer and Diabroctis mimas (Bione et al., 2005a; Cabral-de-Mello et al., 2010b; Oliveira et al., 2010). However, the subfamily Scarabaeinae shows different patterns of CH quantity and distribution, including species that conserve the pattern that is probably the ancestral condition of the family Scarabaeidae, with a predominance of centromeric/pericentromeric heterochromatin blocks (Moura et al., 2003; Wilson and Angus, 2004, 2005; Bione et al., 2005b). "
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    ABSTRACT: The genus Phanaeus is included in the tribe Phanaeini, one of the most diverse tribes within the subfamily Scarabaeinae in terms of chromosomal characteristics. However, so far the species of this genus were not studied with differential cytogenetic techniques, limiting any inference of the probable mechanisms responsible for this diversity. In this work, several techniques were applied with the aim of cytogenetically characterizing two Phanaeus species. The karyotype found for Phanaeus (Notiophanaeus) chalcomelas was 2n = 12, neo-XY, and that of P. (N.) splendidulus was 2n = 20, Xyp, considered primitive for the family Scarabaeidae. The chromosomes of both species showed a high amount of constitutive heterochromatin (CH), with blocks rich in base pairs GC (CMA3 (+)). Moreover, in P. (N.) chalcomelas the marks revealed by C-banding and fluorochrome staining were different in size, showing CH variability. Sites of 18S ribosomal DNA (rDNA) were identified in one autosomal pair of P. (N.) chalcomelas and in five autosomal pairs of P. (N.) splendidulus. On the other hand, only one autosomal pair exhibited 5S rDNA sequences in these species. The results suggest that the karyotype differentiation of the Phanaeus species studied here involved pericentric inversions and centric fusions, as well as mechanisms related to amplification and dispersion of CH and rDNA sequences.
    Genetics and Molecular Biology 09/2013; 36(3):341-346. DOI:10.1590/S1415-47572013005000031 · 1.20 Impact Factor
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    • "Interestingly in species whose the relationship in position for heterochromatic blocks and major rDNA was possible to determine it was observed a general pattern for non co-localization in some representatives without dispersion for these two chromosomal markers, such as in Dichotomius [28]. In species with spreading of these elements in general they were co-located, such as in Deltochilum and Coprophaneus [26,27]. Our results indicate that the same evolutionary forces might be acting on these two components of the Scarabaeinae genome, resulting in the spreading of the major rDNA clusters along with heterochromatin. "
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    ABSTRACT: Scarabaeinae beetles show a high level of macro-chromosomal variability, although the karyotypic organization of heterochromatin and multigene families (rDNAs and histone genes) is poorly understood in this group. To better understand the chromosomal organization and evolution in this group, we analyzed the karyotypes, heterochromatin distribution and chromosomal locations of the rRNAs and histone H3 genes in beetles belonging to eight tribes from the Scarabaeinae subfamily (Coleoptera, Scarabaeidae). The number of 18S rRNA gene (a member of the 45S rDNA unit) sites varied from one to 16 and were located on the autosomes, sex chromosomes or both, although two clusters were most common. Comparison of the 45S rDNA cluster number and the diploid numbers revealed a low correlation value. However, a comparison between the number of 45S rDNA sites per genome and the quantity of heterochromatin revealed (i) species presenting heterochromatin restricted to the centromeric/pericentromeric region that contained few rDNA sites and (ii) species with a high quantity of heterochromatin and a higher number of rDNA sites. In contrast to the high variability for heterochromatin and 45S rDNA cluster, the presence of two clusters (one bivalent cluster) co-located on autosomal chromosomes with the 5S rRNA and histone H3 genes was highly conserved. Our results indicate that the variability of the 45S rDNA chromosomal clusters is not associated with macro-chromosomal rearrangements but are instead related to the spread of heterochromatin. The data obtained also indicate that both heterochromatin and the 45S rDNA loci could be constrained by similar evolutionary forces regulating spreading in the distinct Scarabaeinae subfamily lineages. For the 5S rRNA and the histone H3 genes, a similar chromosomal organization could be attributed to their association/co-localization in the Scarabaeinae karyotypes. These data provide evidence that different evolutionary forces act at the heterochromatin and the 45S rDNA loci compared to the 5S rRNA and histone H3 genes during the evolution of the Scarabainae karyotypes.
    BMC Genetics 10/2011; 12:88. DOI:10.1186/1471-2156-12-88 · 2.40 Impact Factor
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