Adhesion force measurements on the two wax layers of the waxy zone in Nepenthes alata pitchers

The wax coverage of the waxy zone in Nepenthes alata pitchers consists of two clearly distinguishable layers, designated the upper and lower wax layers. Since these layers were reported to reduce insect attachment, they were considered to have anti-adhesive properties. However, no reliable adhesion tests have been performed with these wax layers. In this study, pull-off force measurements were carried out on both wax layers of the N. alata pitcher and on two reference polymer surfaces using deformable polydimethylsiloxane half-spheres as probes. To explain the results obtained, roughness measurements were performed on test surfaces. Micro-morphology of both surface samples and probes tested was examined before and after experiments. Pull-off forces measured on the upper wax layer were the lowest among surfaces tested. Here, contamination of probes by wax crystals detached from the pitcher surface was found. This suggests that low insect attachment on the upper wax layer is caused primarily by the breaking off of wax crystals from the upper wax layer, which acts as a separation layer between the insect pad and the pitcher surface. High adhesion forces obtained on the lower wax layer are explained by the high deformability of probes and the particular roughness of the substrate.