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Keller A.,University of Wurzburg | Danner N.,University of Wurzburg | Grimmer G.,University of Wurzburg | Ankenbrand M.,University of Wurzburg | And 5 more authors.
Plant Biology

The identification of pollen plays an important role in ecology, palaeo-climatology, honey quality control and other areas. Currently, expert knowledge and reference collections are essential to identify pollen origin through light microscopy. Pollen identification through molecular sequencing and DNA barcoding has been proposed as an alternative approach, but the assessment of mixed pollen samples originating from multiple plant species is still a tedious and error-prone task. Next-generation sequencing has been proposed to avoid this hindrance. In this study we assessed mixed pollen probes through next-generation sequencing of amplicons from the highly variable, species-specific internal transcribed spacer 2 region of nuclear ribosomal DNA. Further, we developed a bioinformatic workflow to analyse these high-throughput data with a newly created reference database. To evaluate the feasibility, we compared results from classical identification based on light microscopy from the same samples with our sequencing results. We assessed in total 16 mixed pollen samples, 14 originated from honeybee colonies and two from solitary bee nests. The sequencing technique resulted in higher taxon richness (deeper assignments and more identified taxa) compared to light microscopy. Abundance estimations from sequencing data were significantly correlated with counted abundances through light microscopy. Simulation analyses of taxon specificity and sensitivity indicate that 96% of taxa present in the database are correctly identifiable at the genus level and 70% at the species level. Next-generation sequencing thus presents a useful and efficient workflow to identify pollen at the genus and species level without requiring specialised palynological expert knowledge. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands. Source

Luken D.J.,Laves Institute For Bienenkunde | Janke M.,Laves Institute For Bienenkunde | Lienau F.-W.,Laves Institute For Bienenkunde | von der Ohe W.,Laves Institute For Bienenkunde | Forster R.,Bundesamt fur Verbraucherschutz und Lebensmittelsicherheit BVL
Journal fur Verbraucherschutz und Lebensmittelsicherheit

In the LAVES Institute for Apiculture honey bee brood has been reared in the laboratory according to the method of Aupinel et al. (Bull Insectol 58:107-111, 2005) for studies on the effects of plant protection products as well as for further improvement of the method since 2008. The introduced methodological changes have a positive effect on the number of larvae available for experimental purposes. Furthermore, the treatment with MBC-solution (methyl-benzethonium chloride) could be replaced by pasteurization, which is to be viewed positively from a health perspective for the personnel but also in terms of possible synergistic effects with active ingredients. © 2012 Bundesamt für Verbraucherschutz und Lebensmittelsicherheit (BVL). Source

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