Guided droplet transport on synthetic slippery surfaces inspired by a pitcher plant.

We show how anisotropic, grooved features facilitate the trapping and directed transport of droplets on lubricated, liquid-shedding surfaces. Capillary action pins droplets to topographic surface features, enabling transport along the feature while inhibiting motion across (or detachment from) the feature. We demonstrate the robustness of this capillary-based mechanism for directed droplet transport on slippery surfaces by combining experiments on synthetic, lubricant-infused surfaces with observations on the natural trapping surface of a carnivorous pitcher plant. Controlling liquid navigation on synthetic surfaces promises to unlock significant potential in droplet-based technologies. Our observations also offer novel insight into the evolution of the Nepenthes pitcher plant, indicating that the 'pitfall' trapping mechanism is enhanced by the lubricant-infused, macroscopic grooves on the slippery peristome surface, which guide prey into the trap in a way that is more tightly controlled than previously considered.

The cobra's tongue: Rethinking the function of the "fishtail appendage" on the pitcher plant Darlingtonia californica.

PREMISE OF STUDY: Carnivorous pitcher plants employ a variety of putative adaptations for prey attraction and capture. One example is the peculiar forked "fishtail appendage", a foliar structure widely presumed to function as a prey attractant on adult leaves of Darlingtonia californica (Sarraceniaceae). This study tests the prediction that the presence of the appendage facilitates prey capture and can be considered an example of an adaptation to the carnivorous syndrome. METHODS: In a field experiment following a cohort of Darlingtonia leaves over their growing season, before the pitcher traps opened, the fishtail appendages from half of the leaves were removed. Additionally, all appendages were removed from every plant at two small, isolated populations. After 54 and 104 d, prey items were collected to determine whether differences in prey composition and biomass existed between experimental and unmanipulated control leaves. KEY RESULTS: Removal of the fishtail appendage did not reduce pitcher leaves' prey biomass nor alter their prey composition at either the level of individual leaves or entire populations. Fishtail appendages on plants growing in shaded habitats contained significantly greater chlorophyll concentrations than those on plants growing in full sun. CONCLUSIONS: These results call into question the longstanding assumption that the fishtail appendage on Darlingtonia is an adaptation critical for the attraction and capture of prey. I suggest alternative evolutionary explanations for the role of the fishtail structure and repropose a hypothesis on the mutualistic nature of pitcher plant-arthropod trophic interactions.

Fenestration: a window of opportunity for carnivorous plants.

A long-standing but controversial hypothesis assumes that carnivorous plants employ aggressive mimicry to increase their prey capture success. A possible mechanism is that pitcher plants use aggressive mimicry to deceive prey about the location of the pitcher's exit. Specifically, species from unrelated families sport fenestration, i.e. transparent windows on the upper surfaces of pitchers which might function to mimic the exit of the pitcher. This hypothesis has not been evaluated against alternative hypotheses predicting that fenestration functions to attract insects from afar. By manipulating fenestration, we show that it does not increase the number of Drosophila flies or of two ant species entering pitchers in Sarracenia minor nor their retention time or a pitcher's capture success. However, fenestration increased the number of Drosophila flies alighting on the pitcher compared with pitchers of the same plant without fenestration. We thus suggest that fenestration in S. minor is not an example of aggressive mimicry but rather functions in long-range attraction of prey. We highlight the need to evaluate aggressive mimicry relative to alternative concepts of plant-animal communication.

Isolation of viable multicellular glands from tissue of the carnivorous plant, Nepenthes.

Many plants possess specialized structures that are involved in the production and secretion of specific low molecular weight compounds and proteins. These structures are almost always localized on plant surfaces. Among them are nectaries or glandular trichomes. The secreted compounds are often employed in interactions with the biotic environment, for example as attractants for pollinators or deterrents against herbivores. Glands that are unique in several aspects can be found in carnivorous plants. In so-called pitcher plants of the genus Nepenthes, bifunctional glands inside the pitfall-trap on the one hand secrete the digestive fluid, including all enzymes necessary for prey digestion, and on the other hand take-up the released nutrients. Thus, these glands represent an ideal, specialized tissue predestinated to study the underlying molecular, biochemical, and physiological mechanisms of protein secretion and nutrient uptake in plants. Moreover, generally the biosynthesis of secondary compounds produced by many plants equipped with glandular structures could be investigated directly in glands. In order to work on such specialized structures, they need to be isolated efficiently, fast, metabolically active, and without contamination with other tissues. Therefore, a mechanical micropreparation technique was developed and applied for studies on Nepenthes digestion fluid. Here, a protocol is presented that was used to successfully prepare single bifunctional glands from Nepenthes traps, based on a mechanized microsampling platform. The glands could be isolated and directly used further for gene expression analysis by PCR techniques after preparation of RNA.

The use of light in prey capture by the tropical pitcher plant Nepenthes aristolochioides.

Nepenthes pitcher plants deploy tube-shaped pitchers to catch invertebrate prey; those of Nepenthes aristolochioides possess an unusual translucent dome. The hypothesis was tested that N. aristolochioides pitchers operate as light traps, by quantifying prey capture under three shade treatments. Flies are red-blind, with visual sensitivity maxima in the UV, blue, and green wavebands. Red celluloid filters were used to reduce the transmission of these wavebands into the interior of the pitchers. Those that were shaded at the rear showed a 3-fold reduction in Drosophila caught, relative to either unshaded control pitchers, or pitchers that were shaded at the front. Thus, light transmitted through the translucent dome is a fundamental component of N. aristolochioides' trapping mechanism.

Functional characterization of a class III acid endochitinase from the traps of the carnivorous pitcher plant genus, Nepenthes.

Carnivory in plants is an adaptation strategy to nutrient-poor environments and soils. Carnivorous plants obtain some additional mineral nutrients by trapping and digesting prey; the genus Nepenthes is helped by its specialized pitcher traps. To make the nutrients available, the caught prey needs to be digested, a process that requires the concerted activity of several hydrolytic enzymes. To identify and investigate the various enzymes involved in this process, fluid from Nepenthes traps has been analysed in detail. In this study, a novel type of Nepenthes endochitinase was identified in the digestion fluid of closed pitchers. The encoding endochitinase genes have been cloned from eight different Nepenthes species. Among these, the deduced amino acid sequence similarity was at least 94.9%. The corresponding cDNA from N. rafflesiana was heterologously expressed, and the purified protein, NrChit1, was biochemically characterized. The enzyme, classified as a class III acid endochitinase belonging to family 18 of the glycoside hydrolases, is secreted into the pitcher fluid very probably due to the presence of an N-terminal signal peptide. Transcriptome analyses using real-time PCR indicated that the presence of prey in the pitcher up-regulates the endochitinase gene not only in the glands, which are responsible for enzyme secretion, but at an even higher level, in the glands' surrounding tissue. These results suggest that in the pitchers' tissues, the endochitinase as well as other proteins from the pitcher fluid might fulfil a different, primary function as pathogenesis-related proteins.

Nutritional benefit from leaf litter utilization in the pitcher plant Nepenthes ampullaria.

The pitcher plant Nepenthes ampullaria has an unusual growth pattern, which differs markedly from other species in the carnivorous genus Nepenthes. Its pitchers have a reflexed lid and sit above the soil surface in a tighly packed 'carpet'. They contain a significant amount of plant-derived materials, suggesting that this species is partially herbivorous. We tested the hypothesis that the plant benefits from leaf litter utilization by increased photosynthetic efficiency sensu stricto cost/benefit model. Stable nitrogen isotope abundance indicated that N. ampullaria derived around 41.7 ± 5.5% of lamina and 54.8 ± 7.0% of pitcher nitrogen from leaf litter. The concentrations of nitrogen and assimilation pigments, and the rate of net photosynthesis (A(N)), increased in the lamina as a result of feeding, but did not increase in the trap. However, maximal (F(v) /F(m)) and effective photochemical quantum yield of photosystem II (Φ(PSII)) were unaffected. Our data indicate that N. ampullaria benefits from leaf litter utilization and our study provides the first experimental evidence that the unique nitrogen sequestration strategy of N. ampullaria provides benefits in term of photosynthesis and growth.

Traps of carnivorous pitcher plants as a habitat: composition of the fluid, biodiversity and mutualistic activities.

BACKGROUND: Carnivorous pitcher plants (CPPs) use cone-shaped leaves to trap animals for nutrient supply but are not able to kill all intruders of their traps. Numerous species, ranging from bacteria to vertrebrates, survive and propagate in the otherwise deadly traps. This paper reviews the literature on phytotelmata of CPPs. PITCHER: Fluid as a Habitat The volumes of pitchers range from 0·2 mL to 1·5 L. In Nepenthes and Cephalotus, the fluid is secreted by the trap; the other genera collect rain water. The fluid is usually acidic, rich in O(2) and contains digestive enzymes. In some taxa, toxins or detergents are found, or the fluid is extremely viscous. In Heliamphora or Sarracenia, the fluid differs little from pure water. INQUILINE: Diversity Pitcher inquilines comprise bacteria, protozoa, algae, fungi, rotifers, crustaceans, arachnids, insects and amphibia. The dominant groups are protists and Dipteran larvae. The various species of CPPs host different sets of inquilines. Sarracenia purpurea hosts up to 165 species of inquilines, followed by Nepenthes ampullaria with 59 species, compared with only three species from Brocchinia reducta. Reasons for these differences include size, the life span of the pitcher as well as its fluid. MUTUALISTIC: Activities Inquilines closely interact with their host. Some live as parasites, but the vast majority are mutualists. Beneficial activities include secretion of enzymes, feeding on the plant's prey and successive excretion of inorganic nutrients, mechanical break up of the prey, removal of excessive prey and assimilation of atmospheric N(2). CONCLUSIONS: There is strong evidence that CPPs influence their phytotelm. Two strategies can be distinguished: (1) Nepenthes and Cephalotus produce acidic, toxic or digestive fluids and host a limited diversity of inquilines. (2) Genera without efficient enzymes such as Sarracenia or Heliamphora host diverse organisms and depend to a large extent on their symbionts for prey utilization.

Slippery surfaces of pitcher plants: Nepenthes wax crystals minimize insect attachment via microscopic surface roughness.

Pitcher plants of the genus Nepenthes efficiently trap and retain insect prey in highly specialized leaves. Besides a slippery peristome which inhibits adhesion of insects they employ epicuticular wax crystals on the inner walls of the conductive zone of the pitchers to hamper insect attachment by adhesive devices. It has been proposed that the detachment of individual crystals and the resulting contamination of adhesive organs is responsible for capturing insects. However, our results provide evidence in favour of a different mechanism, mainly based on the stability and the roughness of the waxy surface. First, we were unable to detect a large quantity of crystal fragments on the pads of insects detached from mature pitcher surfaces of Nepenthes alata. Second, investigation of the pitcher surface by focused ion beam treatment showed that the wax crystals form a compact 3D structure. Third, atomic force microscopy of the platelet-shaped crystals revealed that the crystals are mechanically stable, rendering crystal detachment by insect pads unlikely. Fourth, the surface profile parameters of the wax layer showed striking similarities to those of polishing paper with low grain size. By measuring friction forces of insects on this artificial surface we demonstrate that microscopic roughness alone is sufficient to minimize insect attachment. A theoretical model shows that surface roughness within a certain length scale will prevent adhesion by being too rough for adhesive pads but not rough enough for claws.

Trap geometry in three giant montane pitcher plant species from Borneo is a function of tree shrew body size.

*Three Bornean pitcher plant species, Nepenthes lowii, N. rajah and N. macrophylla, produce modified pitchers that 'capture' tree shrew faeces for nutritional benefit. Tree shrews (Tupaia montana) feed on exudates produced by glands on the inner surfaces of the pitcher lids and defecate into the pitchers. *Here, we tested the hypothesis that pitcher geometry in these species is related to tree shrew body size by comparing the pitcher characteristics with those of five other 'typical' (arthropod-trapping) Nepenthes species. *We found that only pitchers with large orifices and lids that are concave, elongated and oriented approximately at right angles to the orifice capture faeces. The distance from the tree shrews' food source (that is, the lid nectar glands) to the front of the pitcher orifice precisely matches the head plus body length of T. montana in the faeces-trapping species, and is a function of orifice size and the angle of lid reflexion. *Substantial changes to nutrient acquisition strategies in carnivorous plants may occur through simple modifications to trap geometry. This extraordinary plant-animal interaction adds to a growing body of evidence that Nepenthes represents a candidate model for adaptive radiation with regard to nitrogen sequestration strategies.

Differential expressed protein in developing stages of Nepenthes gracilis Korth. pitcher.

Nepenthes gracilis Korth. is a member of carnivorous plants in family Nepenthaceae. The plants have beautiful and economically important pitchers. It is interesting to study the protein(s) correlated with the pitcher. Crude proteins were extracted from leaf, leaf with developing pitcher and developed pitcher of the same plant and analyzed by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Two protein bands with molecular weights of 42.7 and 38 kDa were obtained from young leaf and leaf with developing pitcher, respectively. The 42.7 kDa protein was identified as phosphoglycerate kinase (PGK) by Liquid Chromatography Mass Spectrometry (LC-MS/MS), but the 38 kDa band is an unknown protein. Both proteins were differentially expressed in each developing stage of the pitcher, thus may be powerful candidates play role in development pathway of leaf and pitcher.

Longevity, lignin content and construction cost of the assimilatory organs of Nepenthes species.

BACKGROUND AND AIMS: This study examined level of causal relationships amongst functional traits in leaves and conjoint pitcher cups of the carnivorous Nepenthes species. METHODS: Physico-chemical properties, especially lignin content, construction costs, and longevity of the assimilatory organs (leaf and pitcher) of a guild of lowland Nepenthes species inhabiting heath and/or peat swamp forests of Brunei, northern Borneo were determined. KEY RESULTS: Longevity of these assimilatory organs was linked significantly to construction cost, lignin content and structural trait of tissue density, but these effects are non-additive. Nitrogen and phosphorus contents (indicators of Rubisco and other photosynthetic proteins), were poor predictors of organ longevity and construction cost, suggesting that a substantial allocation of biomass of the assimilatory organs in Nepenthes is to structural material optimized for prey capture, rigidity and escape from biotic and abiotic stresses rather than to light interception. Leaf payback time - a measure of net carbon revenue - was estimated to be 48-60 d. This is in line with the onset of substantial mortality by 2-3 months of tagged leaves in many of the Nepenthes species examined. However, this is a high ratio (i.e. a longer minimum payback time) compared with what is known for terrestrial, non-carnivorous plants in general (5-30 d). CONCLUSIONS: It is concluded that the leaf trait bivariate relationships within the Nepenthes genus, as in other carnivorous species (e.g. Sarraceniaceae), is substantially different from the global relationship documented in the Global Plant Trait Network.

Nutrient limitation and morphological plasticity of the carnivorous pitcher plant Sarracenia purpurea in contrasting wetland environments.

* Plasticity of leaf nutrient content and morphology, and macronutrient limitation were examined in the northern pitcher plant, Sarracenia purpurea subsp. purpurea, in relation to soil nutrient availability in an open, neutral pH fen and a shady, acidic ombrotrophic bog, over 2 yr following reciprocal transplantation of S. purpurea between the wetlands. * In both wetlands, plants were limited by nitrogen (N) but not phosphorus (P) (N content < 2% DW(-1), N : P < 14) but photosynthetic quantum yields were high (F(V)/F(M) > 0.79). Despite carnivory, leaf N content correlated with dissolved N availability to plant roots (leaf N vs , r(2) = 0.344, P < 0.0001); carnivorous N acquisition did not apparently overcome N limitation. * Following transplantation, N content and leaf morphological traits changed in new leaves to become more similar to plants in the new environment, reflecting wetland nutrient availability. Changes in leaf morphology were faster when plants were transplanted from fen to bog than from bog to fen, possibly reflecting a more stressful environment in the bog. * Morphological plasticity observed in response to changes in nutrient supply to the roots in natural habitats complements previous observations of morphological changes with experimental nutrient addition to pitchers.