A1 Refereed original research article in a scientific journal
Molecular identification of cryptic bumblebee species from degraded samples using PCR-RFLP approach
Authors: Vesterlund SR, Sorvari J, Vasemagi A
Publisher: WILEY-BLACKWELL
Publication year: 2014
Journal: Molecular Ecology Resources
Journal name in source: MOLECULAR ECOLOGY RESOURCES
Journal acronym: MOL ECOL RESOUR
Volume: 14
Issue: 1
First page : 122
Last page: 126
Number of pages: 5
ISSN: 1755-098X
DOI: https://doi.org/10.1111/1755-0998.12168(external)
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 (446bp) 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 (1064bp) 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.