Cytochrome c oxidase I primers for corbiculate bees: DNA barcode and mini‐barcode

Abstract

Bees (Apidae), of which there are more than 19 900 species, are extremely important for ecosystem services and economic purposes, so taxon identity is a major concern. The goal of this study was to optimize the DNA barcode technique based on the Cytochrome c oxidase (COI) mitochondrial gene region. This approach has previously been shown to be useful in resolving taxonomic inconsistencies and for species identification when morphological data are poor. Specifically, we designed and tested new primers and standardized PCR conditions to amplify the barcode region for bees, focusing on the corbiculate Apids. In addition, primers were designed to amplify small COI amplicons and tested with pinned specimens. Short barcode sequences were easily obtained for some Bombus century-old museum specimens and shown to be useful as mini-barcodes. The new primers and PCR conditions established in this study proved to be successful for the amplification of the barcode region for all species tested, regardless of the conditions of tissue preservation. We saw no evidence of Wolbachia or numts amplification by these primers, and so we suggest that these new primers are of broad value for corbiculate bee identification through DNA barcode.

Morphological, chemical, and molecular analyses differentiate populations of the subterranean nesting stingless bee Mourella caerulea (Apidae: Meliponini)

“…We extracted total DNA from the middle leg of one individual per nest (sampling site in the case of Bagé) according to the methodology proposed by Walsh et al (1991). We performed a PCR using conditions previously described (Françoso and Arias 2013;Simon et al 1994). For the PCR, it was used 5 μl of DNA, 2 μl of each primer (10 pmol), 25 μl of 2× Promega PCRMaster Mix [Taq-DNA polymerase (0.05 units/μl), 2× reaction buffer, 400 μM of each dNTP, and 3 mM of MgCl 2 ], and 16 μl of ultra-pure water.…”

The marker choice: Unexpected resolving power of an unexplored CO1 region for layered DNA barcoding approaches

“…[29]) and insects (e.g. [30]). More interestingly, in most studies using this partition of the CO1 gene, taxon specific primers have been used instead of the universal primers established by Folmer et al in 1994 [29–32].…”

Patterns of sexual size dimorphism in stingless bees: Testing Rensch’s rule and potential causes in highly eusocial bees (Hymenoptera: Apidae, Meliponini)

“…To remove the phylogenetic effects from the analysis of data, we built a phylogeny of Meliponini species mainly based on the genetic information available on the GenBank. This genetic information involved partial sequences of two mitochondrial loci (cytochrome oxidase subunit I, COI; and 16S ribosomal RNA, 16S) and four nuclear loci (arginine kinase, ArgK; elongation factor 1 alpha, EF‐1α; long‐wavelength rhodopsin, LWR; and 28S ribosomal RNA, 28S) that were used in previous phylogenetic studies in stingless bees (Françoso & Arias, ; Halcroft et al, ; May‐Itza, Quezada‐Euán, Medina, Enríquez, & De la Rúa, ; Ramírez et al, ; Rasmussen & Cameron, ; Ruiz, May‐Itza, Quezada‐Euán, & De la Rúa, ). Particularly, COI sequences for three Meliponini species ( Frieseomelitta nigra , Lestrimelitta niitkib , and Partamona bilineata ) were provided by the Canadian Barcode of Life Network.…”

“…To remove the phylogenetic effects from the analysis of data, we built a phylogeny of Meliponini species mainly based on the genetic information available on the GenBank. This genetic information involved partial sequences of two mitochondrial loci (cytochrome oxidase subunit I, COI; and 16S ribosomal RNA, 16S) and four nuclear loci (arginine kinase, ArgK; elongation factor 1 alpha, EF-1α; long-wavelength rhodopsin, LWR; and 28S ribosomal RNA, 28S) that were used in previous phylogenetic studies in stingless bees (Françoso & Arias, 2013;Halcroft et al, 2016 we first homologated the taxonomic information found in the literature search with the updated revision by Camargo and Pedro (2007) and Rasmussen and Cameron (2010). We also retrieved from GenBank genetic information of five corbiculate bee species (Apis…”