Chromosomal mapping of repetitive DNA in Melipona seminigra merrillae Cockerell, 1919 (Hymenoptera, Apidae, Meliponini)

Abstract

Melipona Illiger, 1806 is represented by 74 known species of stingless bees, distributed throughout the Neotropical region. Cytogenetically it is the most studied stingless bee genus of the tribe Meliponini. Member species are divided in two groups based on the volume of heterochromatin. This study aim was to analyze the composition and organization of chromatin of the stingless bee subspecies Melipona seminigra merrillae Cockerell, 1919 using classical and molecular cytogenetic techniques, so contributing to a better understanding of the processes of chromosomal changes within the genus. We confirm that M. seminigra merrillae has a chromosome number of 2n = 22 and n = 11, results that differ from those reported for the genus in the absence of B chromosomes. The heterochromatic pattern revealed a karyotype composed of chromosomes with a high heterochromatin content, which makes it difficult to visualize the centromere. Silver nitrate impregnation (Ag-NOR) showed transcriptionally active sites on the second chromosomal pair. Staining of base-specific fluorophores DAPI-CMA3 indicated a homogeneous distribution of intensely DAPI-stained heterochromatin, while CMA3 markings appeared on those terminal portions of the chromosomes corresponding to euchromatin. Similar to Ag-NOR, fluorescence in situ hybridization (FISH) with 18S ribosomal DNA probe revealed distinct signals on the second pair of chromosomes. Microsatellite mapping (GA)15 showed markings distributed in euchromatic regions, while mapping with (CA)15 showed marking patterns in heterochromatic regions, together with a fully marked chromosome pair. Microsatellite hybridization, both in heterochromatic and euchromatic regions, may be related to the activity of transposable elements. These are capable of forming new microsatellites that can be dispersed and amplified in different regions of the genome, demonstrating that repetitive sequences can evolve rapidly, thus resulting in within-genus diversification.