Cytogenetic analysis of two related Deltochilum (Coleoptera, Scarabaeidae) species: Diploid number reduction, extensive heterochromatin addition and differentiation
ABSTRACT 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.
SourceAvailable from: Diogo Cabral-de-Mello[Show abstract] [Hide abstract]
ABSTRACT: The use of fluorescent in situ hybridization (FISH) has allowed the generation of data concerning the genome structure and chromosomal organization and differentiation of diverse eukaryote organisms. This technique guided to a revolution in the cytogenetic and permitted a more clear visualization of specific DNA sequences onto chromosomes, thus generating detailed physical chromosome maps of species. For studies of genome organization and karyotype ongoing in animals, the most applied sequences are the repeated elements. These elements have been obtained from the genome of distinct species mainly through the use of polymerase chain reaction (PCR) and enzymatic restriction, and have been used for chromosome identification, establishment of chromosomal rearrangements, studies of B and sex chromosomes origin and evolution, and genome organization. Here we summarize the recent advances in the application of the FISH technology in studies of fish and insect chromosomes, under the focus of understanding the organization and evolution of their karyotypes and genomes.
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ABSTRACT: Triatoma infestans, one of the most important vectors of Trypanosoma cruzi, is very interesting model, because it shows large interpopulation variation in the amount and distribution of heterochromatin. This polymorphism involved the three large pairs up to almost all autosomal pairs, including the sex chromosomes. To understand the dynamics of heterochromatin variation in T. infestans, we isolated the AT-rich satDNA portion of this insect using reassociation kinetics (C0t), followed by cloning, sequencing and FISH. After chromosome localization, immunolabeling with anti-5-methylcytosine, anti-H4K5ac and anti-H3K9me2 antibodies was performed to determine the functional characteristics of heterochromatin. The results allowed us to reorganize the karyotype of T. infestans in accordance with the distribution of the families of repetitive DNA using seven different markers. We found that two arrays with lengths of 79 bp and 33 bp length have a strong relationship with transposable element sequences, suggesting that these two families of satDNA probably originated from Polintons. The results also allowed us to identify at least four chromosome rearrangements involved in the amplification/dispersion of AT-rich satDNA of T. infestans. These data should be very useful in new studies including those examining the cytogenomic and population aspects of this very important species of insect.Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 04/2014; 23. DOI:10.1016/j.meegid.2014.01.035 · 3.26 Impact Factor
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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. DOI:10.1186/1755-8166-6-54 · 2.66 Impact Factor