Development and regeneration ability of the wax coverage in Nepenthes alata pitchers: a cryo-SEM approach

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Abstract

The morphogenesis of the composite epicuticular wax coverage and regeneration ability of the upper wax layer in Nepenthes alata pitchers were studied using a cryo-scanning electron microscopy. Examination of pitchers of different ages revealed six stages in the wax coverage development. In the first stage, wax crystals resemble those found recently in mature pitches of N. dicksoniana and N. ventricosa. Platelets of the upper wax layer originate from broadened tips of stalks during the last developmental stage. Contrary to previous hypotheses, we found that wax crystals of both layers as well as the stalks connecting them are oriented perpendicularly to the pitcher wall. No changes in the height of the wax coverage were detected in 4–8 weeks after mechanical removal of the upper wax layer from mature pitchers on plants. This indicates that the wax coverage in N. alata pitchers is unable to regenerate.

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Superhydrophobic and superhydrophilic plant surfaces: an inspiration for biomimetic materials.

Kerstin Koch, Wilhelm Barthlott (2009)

The diversity of plant surface structures, evolved over 460 million years, has led to a large variety of highly adapted functional structures. The plant cuticle provides structural and chemical modifications for surface wetting, ranging from superhydrophilic to superhydrophobic. In this paper, the structural basics of superhydrophobic and superhydrophilic plant surfaces and their biological functions are introduced. Wetting in plants is influenced by the sculptures of the cells and by the fine structure of the surfaces, such as folding of the cuticle, or by epicuticular waxes. Hierarchical structures in plant surfaces are shown and further types of plant surface structuring leading to superhydrophobicity and superhydrophilicity are presented. The existing and potential uses of superhydrophobic and superhydrophilic surfaces for self-cleaning, drag reduction during moving in water, capillary liquid transport and other biomimetic materials are shown.

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Movement and regeneration of epicuticular waxes through plant cuticles.

W Barthlott, Kerstin Koch, C Neinhuis (2001)

Regeneration of plant epicuticular waxes was studied in 24 plant species by high-resolution scanning electron microscopy. According to their regeneration behaviour, four groups could be distinguished: (i) regeneration occurs at all stages of development; (ii) regeneration occurs only during leaf expansion; (iii) regeneration occurs only in fully developed leaves; (iv) plants were not able to regenerate wax. Wax was removed from the leaves with water-based glue and a liquid polymer, i.e. water-based polyurethane dispersion. In young leaves these coverings could not be removed without damaging the leaves. After a few days, waxes appeared on the surface of these polymer films, which still adhered to the leaves. It is concluded that waxes move through the cuticle in a process similar to steam distillation. This hypothesis could be further substantiated in refined in vitro experiments. Wax isolated from Eucalyptus globulus was applied to a filter paper, subsequently covered with a liquid polymer and fixed onto a diffusion chamber filled with water. The diffusion chamber was put into a desiccator. After 8-10 days at room temperature, crystals similar in dimensions and shape to in situ crystals appeared on the surface of the polyurethane film. This indicates that waxes in molecular dimensions move together with the water vapor that permeates through the polymer membrane. Based on these results, we propose a new and simple hypothesis for the mechanism of wax movement: the molecules that finally form the epicuticular wax crystals are moved in the cuticular water current.

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Composite structure of the crystalline epicuticular wax layer of the slippery zone in the pitchers of the carnivorous plant Nepenthes alata and its effect on insect attachment.

S Gorb, A Henrich, N Barbakadze … (2005)

The slippery zone situated below the peristome inside pitchers of most carnivorous plants from the genus Nepenthes is covered with a thick layer of epicuticular wax. This slippery zone is reported to play a crucial role in animal trapping and prey retention. In N. alata, the wax coverage consists of two clearly distinguished layers. These layers differ in their structure, chemical composition and mechanical properties, and they reduce the insect attachment in different ways. The lower layer resembles foam, composed of interconnected membraneous platelets protruding from the surface at acute angles. The upper layer consists of densely placed separate irregular platelets, located perpendicular to the subjacent layer. Crystals of the upper layer bear small stalks, directed downwards and providing connections to the lower layer. These morphological distinctions correlate with differences in the chemical composition of waxes. The compound classes of alkanes, aldehydes, primary alcohols, free fatty acids, esters and triterpenoids occurred in extracts from both wax layers, but in different proportions. Chain length distributions in aliphatics were different in extracts from the lower and the upper wax layers. Waxes of the upper and lower layers exhibited different mechanical properties: wax of the lower layer is harder and stiffer than that of the upper layer. Moreover, crystals of the upper layer are brittle and may be easily exfoliated or broken to tiny pieces. Laboratory experiments using tethered insects showed that both wax layers reduce the attachment force of insects. It is assumed that a decrease in insect attachment on the two distinct wax layers is provided by the two different mechanisms: (1) crystals of the upper wax layer contaminate insects' adhesive pads; (2) the lower wax layer leads to a reduction of the real contact area of insects' feet with the plant surface.

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Journal

Journal ID (nlm-ta): Sci Rep

Journal ID (iso-abbrev): Sci Rep

Title: Scientific Reports

Publisher: Nature Publishing Group

ISSN (Electronic): 2045-2322

Publication date (Electronic): 29 October 2013

Publication date Collection: 2013

Volume: 3

Electronic Location Identifier: 3078

Affiliations

[1 ]Department of Functional Morphology and Biomechanics, Zoological Institute, Kiel University , Am Botanischen Garten 9, 24118 Kiel, Germany

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[a ] egorb@ 123456zoologie.uni-kiel.de

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Publisher Item ID: srep03078

DOI: 10.1038/srep03078

PMC ID: 3810656

PubMed ID: 24165663

SO-VID: ff4f3f6a-e733-4973-bde9-bdae2028d4e3

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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

Development and regeneration ability of the wax coverage in Nepenthes alata pitchers: a cryo-SEM approach

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Most cited references 12

Superhydrophobic and superhydrophilic plant surfaces: an inspiration for biomimetic materials.

Movement and regeneration of epicuticular waxes through plant cuticles.

Composite structure of the crystalline epicuticular wax layer of the slippery zone in the pitchers of the carnivorous plant Nepenthes alata and its effect on insect attachment.

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