Abstract
Fluorescence in situ hybridization (FISH) allows identification of particular chromosomes and their rearrangements. Using FISH with signal enhancement via antibody amplification and enzymatically catalysed reporter deposition, we evaluated applicability of universal cytogenetic markers, namely 18S and 5S rDNA genes, U1 and U2 snRNA genes, and histone H3 genes, in the study of the karyotype evolution in moths and butterflies. Major rDNA underwent rather erratic evolution, which does not always reflect chromosomal changes. In contrast, the hybridization pattern of histone H3 genes was well conserved, reflecting the stable organisation of lepidopteran genomes. Unlike 5S rDNA and U1 and U2 snRNA genes which we failed to detect, except for 5S rDNA in a few representatives of early diverging lepidopteran lineages. To explain the negative FISH results, we used quantitative PCR and Southern hybridization to estimate the copy number and organization of the studied genes in selected species. The results suggested that their detection was hampered by long spacers between the genes and/or their scattered distribution. Our results question homology of 5S rDNA and U1 and U2 snRNA loci in comparative studies. We recommend the use of histone H3 in studies of karyotype evolution.
Highlights
Fluorescence in situ hybridization (FISH) allows identification of particular chromosomes and their rearrangements
Major rDNA clusters were detected at a terminal position in 22 out of 30 species
We tested whether commonly used cytogenetic markers, namely 18S rDNA, histone H3, 5S rDNA, and U1 and U2 snRNA genes, are applicable and informative for studies of karyotype evolution in Lepidoptera
Summary
Fluorescence in situ hybridization (FISH) allows identification of particular chromosomes and their rearrangements. FISH with the 18S rDNA probe on male pachytene nuclei revealed one chromosomal pair bearing an interstitial cluster which colocalized with a small block of DAPI-positive heterochromatin (Supplementary Fig. S5a).
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