Characterisation of insect and plant origins using DNA extracted from small volumes of bee honey |
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Authors: | Ida Bærholm Schnell Magdalena Fraser Eske Willerslev M Thomas P Gilbert |
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Institution: | 1.Centre for GeoGenetics, Natural History Museum of Denmark,University of Copenhagen,Copenhagen,Denmark;2.Department of Archaeology and Osteology,Gotland University,Visby,Sweden |
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Abstract: | A DNA-based tool was validated that potentially enables the characterisation of both plant and insect of origin of small (approximately
1 ml) samples of bee honey. Using this method, mitochondrial, nuclear and chloroplast DNA (mtDNA, nuDNA, cpDNA) markers were
successfully extracted, PCR amplified, and sequenced from a range of honeys, and the relative amount of plant nuDNA and cpDNA,
and bee mtDNA in the samples was quantified using quantitative real-time PCR. Short, but taxonomically informative lengths
of insect and plant organelle DNA could be routinely recovered from all honey samples tested, and longer organelle, and nuclear
DNA sequences can be recovered from many. The data also enabled preliminary characterisation of the quality of these different
DNA sources in honey. Although the absolute quantity of the different genetic markers varied considerably between sample,
a general trend was observed of insect mtDNA dominating over plant organelle DNA, and with plant nuclear DNA at the lowest
levels. Furthermore there was a clear correlation between the plant DNA content and the success of the PCR assays. To maximise
successful characterisation of samples, future studies are recommended to focus on the use of organelle markers, and limit
the size of PCR amplicons targeted, although with appropriate sample selection and assay optimisation, other approaches may
be possible. |
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