title: Molecular identification of cryptic bumblebee species from degraded samples using PCR-RFLP approach.
abstract: The worldwide decline and local extinctions of bumblebees have raised a need for fast and accurate tools for species identification. Morphological characters are often not sufficient, and molecular methods have been increasingly used for reliable identification of bumblebee species. Molecular methods often require high-quality DNA which makes them less suitable for analysis of low-quality or older samples. We modified the PCR-RFLP protocol for an efficient and cost-effective identification of four bumblebee species in the subgenus Bombus s. str. (B. lucorum, B. terrestris, B. magnus and B. cryptarum). We used a short partial mitochondrial COI fragment (446 bp) and three diagnostic restriction enzymes (Hinf I, Hinc II and Hae III) to identify species from degraded DNA material. This approach allowed us to efficiently determine the correct species from all degraded DNA samples, while only a subset of samples 64.6% (31 of 48) resulted in successful amplification of a longer COI fragment (1064 bp) using the previously described method. This protocol can be applied for conservation and management of bumblebees within this subgenus and is especially useful for fast species identification from degraded samples.
authors.affiliation: Department of Biology, Section of Ecology, University of Turku, Turku, FI-20014, Finland.
authors.forename: S-R
authors.initials: SR
authors.lastname: Vesterlund
authors.forename: J
authors.initials: J
authors.lastname: Sorvari
authors.forename: A
authors.initials: A
authors.lastname: Vasemägi
mesh_terms: D000818:Animals; D001516:Bees / Q000145:classification* / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D003576:Electron Transport Complex IV / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D014481:United States
chemical_list: D004272:DNA, Mitochondrial; D003576:Electron Transport Complex IV; D004262:DNA Restriction Enzymes
keywords: Bombus; COI; PCR-RFLP; conservation; degraded DNA; species identification