Search bioRxiv⌕ Search

Biology subjects

Degirmenci, L.

Publications and source records attributed to Degirmenci, L..

3 recordsLinked to original sources

A fast and easy mass sampling method for social bee crop content

Honey bees (Apis mellifera) are capable of foraging over large distances around their nest and can thus serve as ideal bioindicators for various research questions. While foraging for nectar, they collect a variety of pollutants such as environmental toxins and agrochemicals, which can be traced in their digestive tract. Likewise, naturally occurring compounds in a bees crop content (e.g., different sugars and pollen grains) can be analyzed to assess the foraging preferences of bees or to study the species composition of flowering plants in the respective environment. Regardless of the research question, a precise, reproducible, and efficient method for examining pollinators crop contents is needed to fully exploit the honey bees potential as a bioindicator. Existing methods are often insufficiently precise with respect to the recovered volume and content, too time-consuming or lack the opportunity to store samples flexibly. Consequently, they do not meet the requirements of large-scale sampling commonly needed in scientific field work. We present a novel, rapid and scalable method for fast and easy mass sampling of honey bee crop contents which holds potential for adaptation to other pollinator taxa such as bumblebees. With this method individuals can be collected in the field and stored prior to sample processing. Crop contents can be obtained later on by centrifuging the entire individual, thereby decoupling sample collection from further analysis. Afterwards, insect samples and extracted fluids can be stored again, enabling subsequent analysis in an efficient and time-saving manner. We demonstrate that crop contents of previously fed animals can be obtained precisely in terms of both volume and composition and present an example of application in honey bee foraging ecology as a proof of concept.

ecology↗

Using a combination of CRISPR/Cas9, behavioural experiments and functional analysis to characterise taste receptors in honeybees

Honeybees (Apis mellifera) need their fine sense of taste to evaluate nectar and pollen sources. Gustatory receptors (Grs) translate taste signals into electrical responses. In vivo experiments primarily demonstrate collective responses of the whole Gr-set, but little is known about the individual impact of receptors. Here, we disentangle for the first time the contributions of three gustatory receptors (AmGr1-3) in sugar sensing of honeybees by combining CRISPR/Cas9 mediated genetic knock-out, electrophysiology and behaviour. AmGr1 responds to multiple sugars. Bees lacking this receptor have a reduced response to sucrose and glucose but not to fructose. AmGr2 acts as co-receptor of AmGr1 in a heterologous expression system, but honeybee knock-out mutants perform normally. Eliminating AmGr3 while preserving AmGr1 and AmGr2 abolishes the perception of fructose but not of sucrose. We thus dissociate the roles of AmGr1, AmGr2 and AmGr3 in honeybee taste perception.

physiology↗

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

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

genetics↗