Microdissection: a tool for bee chromosome studies

Abstract

-The emergence of new molecular biology techniques has provided cytogenetics with tools which allow for the elucidation of questions that classical cytogenetics could not answer. Therefore, the present work standardizes a microdissection protocol for cytogenetic studies in bees. This methodology was first used in these insects and may contribute greatly to studies involving chromosomal rearrangements, heterochromatin composition, B chromosomes and others. For this study, the centromeric region of chromosomes in the stingless bee Tetragonisca fiebrigi was used for probe synthesis. The results demonstrated that the methodology used was efficient, presenting markings in the centromeric regions of several chromosomes. Hybridization in other sites indicates that the probe was able to detect regions that present homology with its sequence. This indicates that the technique is effective to study chromosomal evolution, genome organization and even the origin of B chromosomes.

Similarity of heterochromatic regions in the stingless bees (Hymenoptera: Meliponini) revealed by chromosome painting

“…Moreover, the present results showed that there is a high homology among the heterochromatin regions within each species once the probes generated from one chromosome hybridized in other heterochromatic regions of the original species in both Melipona rufiventris and Tetragonisca fiebrigi. Similar results were found by Fernandes et al (2011) in which probes of heterochromatin centromeric hybridized in many chromosomes.…”

“…The first report using molecular cytogenetics to elucidate the heterochromatin organization in bees was done by Fernandes et al (2011), who analyzed centromeric regions in Tetragonisca fiebrigi. No other works of this approach seem to be available in the literature, so this work is very important to understand the karyotype evolution in bees.…”

“…To check the amount of material needed for the amplification, reactions were tested from one microdissected chromosome, as well as from six chromosomes for both species. The DOP-PCR procedures were carried out under the conditions presented in Fernandes et al (2011).…”

Investigation of Partamona helleri (Apidae, Meliponini) B chromosome origin. An approach by microdissection and whole chromosome painting

“…This technique has been applied successfully to different studies on the evolution of sexual systems and origin of B chromosomes on different fish species (Diniz et al 2008;Henning et al 2008;Silva et al 2009;Voltolin et al 2010). Fernandes et al (2011) constructed specific probes useful to locate some of the regular centromeric regions of the stingless bee Tetragonisca fiebrigi. In this study, we combined microdissection and chromosome painting, C-banding, NOR banding, and base-specific fluorochrome staining to investigate the origin of the chromosome B1 in P. helleri.…”

Cytogenetic Analysis: A New Era of Procedures and Precision

“…More recently, Fernandes et al, (2011) standardized a microdissection protocol for cytogenetic studies of bees. This new technique will permit the generation of extremely specific markings of the genome --markings such as centromeric, telomeric, heterochromatic regions of supernumerary chromosomes, or even the entire chromosome itself.…”