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bioRxiv · 10.1101/2020.03.26.009696

CRISPR/Cas9 mediated mutations as a new tool for studying taste in honeybees

Degirmenci, L.·Geiger, D.·Roge Ferreira, F.·Keller, A.·Krischke, B.·Beye, M.·Steffan-Dewenter, I.·Scheiner, R.
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Abstract

BackgroundHoneybees rely on nectar as their main source of carbohydrates [1]. Sucrose, glucose and fructose are the main components of plant nectars [2] [3]. Intriguingly, honeybees express only three putative sugar receptors (AmGr1, AmGr2 and AmGr3) [4], which is in stark contrast to many other insects and vertebrates. The sugar receptors are only partially characterized [5] [6]. AmGr1 detects different sugars including sucrose and glucose. AmGr2 is assumed to act as a co-receptor only, while AmGr3 is assumedly a fructose receptor. ResultsWe show that honeybee gustatory receptor AmGr3 is highly specialized for fructose perception when expressed in Xenopus oocytes. When we introduced nonsense mutations to the respective AmGr3 gene using CRISPR/Cas9 in eggs of female workers, the resulting mutants displayed almost a complete loss of responsiveness to fructose. In contrast, responses to sucrose were normal. Nonsense mutations introduced by CRISPR/Cas9 in honeybees can thus induce a measurable behavioural change and serve to characterize the function of taste receptors in vivo. ConclusionCRISPR/Cas9 is an excellent novel tool for characterizing honeybee taste receptors in vivo. Biophysical receptor characterisation in Xenopus oocytes and nonsense mutation of AmGr3 in honeybees unequivocally demonstrate that this receptor is highly specific for fructose. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=166 HEIGHT=200 SRC="FIGDIR/small/009696v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@1100f07org.highwire.dtl.DTLVardef@14ac138org.highwire.dtl.DTLVardef@d37468org.highwire.dtl.DTLVardef@7f6abb_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 0C_FLOATNO C_FIG

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Degirmenci, L., Geiger, D., Roge Ferreira, F., Keller, A., Krischke, B., Beye, M., Steffan-Dewenter, I., Scheiner, R.. 2020-03-29. CRISPR/Cas9 mediated mutations as a new tool for studying taste in honeybees. https://doi.org/10.1101/2020.03.26.009696

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