Genetic variability in Melipona quinquefasciata (Hymenoptera, Apidae, Meliponini) from northeastern Brazil determined using the first internal transcribed spacer (ITS1)

Abstract

ABSTRACT. Melipona quinquefasciata is a ground-nesting SouthAmerican stingless bee whose geographic distribution was believed to comprise only the central and southern states of Brazil. We obtained partial sequences (about 500-570 bp) of first internal transcribed spacer (ITS1) nuclear ribosomal DNA from Melipona specimens putatively identified as M. quinquefasciata collected from different localities in northeastern Brazil. To confirm the taxonomic identity of the northeastern samples, specimens from the State of Goiás (Central region of Brazil) were included for comparison. All sequences were deposited in GenBank (accession numbers EU073751-EU073759). The mean nucleotide divergence (excluding sites with insertions/deletions) in the ITS1 sequences was only 1.4%, ranging from 0 to 4.1%. When the sites with insertions/ deletions were also taken into account, sequence divergences varied from 0 to 5.3%. In all pairwise comparisons, the ITS1 sequence from the specimens collected in Goiás was most divergent compared to the ITS1 sequences of the bees from the other locations. However, neighbor-joining phylogenetic analysis showed that all ITS1 sequences from northeastern specimens along with the sample of Goiás were resolved in a single clade with a bootstrap support of 100%. The ITS1 sequencing data thus support the occurrence of M. quinquefasciata in northeast Brazil.

Utility of the ITS1 Region for Phylogenetic Analysis in Stingless Bees: a Case Study of the Endangered Melipona yucatanica Camargo, Moure and Roubik (Hymenoptera: Meliponini).

“…In M. yucatanica ITS1 is within the range of that observed in other Melipona species (from 1391 bp in M. quadrifasciata to 1940 in M. rufiventris, Fernandes-Salomão et al, 2005). The presence of insertions and deletions (indels) in this region is a common feature leading to sequence length variation, as it has been observed between Mexican and Guatemalan M. yucatanica specimens, and among Brazilian populations of the species M. subnitida (Cruz et al, 2006) and within M. quinquefasciata (Pereira et al, 2009).…”

“…The ITS1 datasets used included the complete ITS1 gene for M. yucatanica, M. beecheii and Melipona quadrifasciata Lepeletier and partial sequences of ITS1 3' end for other nine Melipona species obtained from Genbank (FernandesSalomão et al, 2005;Cruz et al, 2006;Pereira et al, 2009 ITS1 sequences were aligned with MAFFT software (http://mafft.cbrc.jp/alignment/server/) using E-INS-i strategy. Parameter of scoring matrix for nucleotide sequences was set to 20PAM/ k = 2 as recommended when aligning related DNA sequences (Katoh & Toh, 2008).…”

“…In particular, in the tribe Meliponini, Fernandes-Salomão et al (2005) determined the complete ITS1 sequence from three Melipona species and used partial sequences of eight species to infer its phylogenetic relationships. At the intraspecific level, the ITS1 region has demonstrated its usefulness for population analysis in the Mexican species Melipona beecheii Bennett (MayItzá et al, 2009; and also in two Brazilian species: Melipona subnitida Ducke (Cruz et al, 2006) and Melipona quinquefasciata Lepeletier (Pereira et al, 2009). …”

Contribuição à filogenia de abelhas Melipona com uso de sequências parciais da região ITS1 do nrDNA

“…A região ITS1 do rDNA do táxon Melipona apresenta um grande potencial para estudos filogenéticos entre espécies (FERNANDES-SALOMÃO et al, 2002), como também pode ajudar a resolver dúvidas taxonômicas relacionadas aos subgêneros deste táxon (FERNANDES-SALOMÃO et al, 2005). Além disso, pode servir para avaliar variações intraespecíficas em populações de abelhas, como demonstrado para M. subnitida (CRUZ et al, 2006) e M. quinquefasciata (PEREIRA et al, 2009).…”

“…Estudos prévios conduzidos por Cruz et al (2006) e Pereira et al (2009) trimetil lamônio -CTAB (FOSTER;TWELL, 1996), realizando as adaptações sugeridas por Cruz et al (2006) e Pereira et al (2009).…”