Effects of arthropod inquilines on growth and reproductive effort among metacommunities of the purple pitcher plant (Sarracenia purpurea var. montana).

Many plant species harbor communities of symbionts that release nutrients used by their host plants. However, the importance of these nutrients to plant growth and reproductive effort is not well understood. Here, we evaluate the relationship between the communities that colonize pitcher plant phytotelmata and the pitcher plants' vegetative growth and flower production to better understand the symbiotic role played by phytotelma communities. We focus on the mountain variety purple pitcher plant (Sarracenia purpurea var. montana), which occurs in small and isolated populations in Western North Carolina. We found that greater symbiont community diversity is associated with higher flower production the following season. We then examined geographic variation in communities and found that smaller plant populations supported less diverse symbiont communities. We relate our observations to patterns of community diversity predicted by community ecology theory.

Transcriptome-wide shift from photosynthesis and energy metabolism upon endogenous fluid protein depletion in young Nepenthes ampullaria pitchers.

Carnivorous pitcher plants produce specialised pitcher organs containing secretory glands, which secrete acidic fluids with hydrolytic enzymes for prey digestion and nutrient absorption. The content of pitcher fluids has been the focus of many fluid protein profiling studies. These studies suggest an evolutionary convergence of a conserved group of similar enzymes in diverse families of pitcher plants. A recent study showed that endogenous proteins were replenished in the pitcher fluid, which indicates a feedback mechanism in protein secretion. This poses an interesting question on the physiological effect of plant protein loss. However, there is no study to date that describes the pitcher response to endogenous protein depletion. To address this gap of knowledge, we previously performed a comparative RNA-sequencing experiment of newly opened pitchers (D0) against pitchers after 3 days of opening (D3C) and pitchers with filtered endogenous proteins (>10 kDa) upon pitcher opening (D3L). Nepenthes ampullaria was chosen as a model study species due to their abundance and unique feeding behaviour on leaf litters. The analysis of unigenes with top 1% abundance found protein translation and stress response to be overrepresented in D0, compared to cell wall related, transport, and signalling for D3L. Differentially expressed gene (DEG) analysis identified DEGs with functional enrichment in protein regulation, secondary metabolism, intracellular trafficking, secretion, and vesicular transport. The transcriptomic landscape of the pitcher dramatically shifted towards intracellular transport and defence response at the expense of energy metabolism and photosynthesis upon endogenous protein depletion. This is supported by secretome, transportome, and transcription factor analysis with RT-qPCR validation based on independent samples. This study provides the first glimpse into the molecular responses of pitchers to protein loss with implications to future cost/benefit analysis of carnivorous pitcher plant energetics and resource allocation for adaptation in stochastic environments.

Inter- and intraspecific competition and shade avoidance in the carnivorous pale pitcher plant in a nutrient-poor savanna.

PREMISE OF THE STUDY: Ecologists generally agree that weak interspecific competition for light contributes to high plant species diversity in ecosystems with nutrient-poor soils. However, the role of competition for light in such ecosystems that are also maintained by fire is poorly understood. I quantified intra- and interspecific competition for light in a fire-maintained nutrient-poor pine savanna by contrasting the effects of conspecific and heterospecific neighbors of the pale pitcher plant, Sarracenia alata. METHODS: Accounting for initial neighbor abundance/aboveground production and initial transplant size, I measured growth and survival of small and large pitcher plant ramets of Sarracenia alata transplanted to the vicinity of natural, undisturbed mixtures of large pitcher plants and their heterospecific neighbors in the field. I tested competition for light and nutrients by clipping conspecific neighbors and by excluding prey from unclipped neighbors of transplants. I tested interspecific competition by uprooting heterospecific neighbors. KEY RESULTS: Plant survivorship increased when conspecific neighbors were clipped and/or starved but not when heterospecific neighbors were uprooted. Small pitcher plants benefited from clipping large conspecific neighbors, suggesting that competition for light was important. Large pitcher plants benefited from excluding prey from their neighbors, with no additional benefit of clipping, suggesting that competition for prey limited their survival. Transplants produced new pitchers that were taller with narrower openings (i.e., shade avoidance) when heterospecific neighbors were left intact but not when conspecifics were unclipped. CONCLUSIONS: Results demonstrate size-dependent intraspecific competition for light and nutrients and interspecific shade avoidance in Sarracenia alata, which could be important to understanding species coexistence in fire-maintained nutrient-poor ecosystems.

Effects of Exyra ridingsii (Lepidoptera: Noctuidae) on Sarracenia flava (Nepenthales: Sarraceniaceae).

Exyra ridingsii (Riley) (Lepidoptera: Noctuidae) is a moth whose obligate host is the pitcher plant Sarracenia flava (L.) (Nepenthales: Sarraceniaceae). The entire life cycle of the moth is completed in the trumpets of this fire-dependent plant that is found throughout the southeastern United States in bogs, long-leaf pine savannas, and pocosins. The purpose of this study was to determine the effects of E. ridingsii on S. flava, including the effect of herbivory on trumpet height in the year subsequent to feeding and whether moths select trumpets for oviposition based on height. Although most forms of herbivory by insects might be expected to have negative effects on plants by reducing photosynthetic abilities, it would be counterproductive for herbivory by E. ridingsii to negatively affect S. flava as this plant is the only possible habitat for E. ridingsii. At each site in selected quadrats, the number of trumpets, trumpet height, trumpet status, number of trumpets in a clump, and number of clumps were recorded. The relationship between height and herbivory was analyzed using a linear model, and a positive correlation was found between height and herbivory. E. ridingsii herbivory had no effect on the next year's growth of S. flava based on a Spearman's correlation. Therefore, we concluded that E. ridingsii has little effect on S. flava populations and has likely evolved to selectively avoid herbivory on more vulnerable, smaller plants.

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.

Oriented cell division shapes carnivorous pitcher leaves of Sarracenia purpurea.

Complex morphology is an evolutionary outcome of phenotypic diversification. In some carnivorous plants, the ancestral planar leaf has been modified to form a pitcher shape. However, how leaf development was altered during evolution remains unknown. Here we show that the pitcher leaves of Sarracenia purpurea develop through cell division patterns of adaxial tissues that are distinct from those in bifacial and peltate leaves, subsequent to standard expression of adaxial and abaxial marker genes. Differences in the orientation of cell divisions in the adaxial domain cause bifacial growth in the distal region and adaxial ridge protrusion in the middle region. These different growth patterns establish pitcher morphology. A computer simulation suggests that the cell division plane is critical for the pitcher morphogenesis. Our results imply that tissue-specific changes in the orientation of cell division underlie the development of a morphologically complex leaf.

Ants provide nutritional and defensive benefits to the carnivorous plant Sarracenia minor.

Ants can have important, but sometimes unexpected, effects on the plants they associate with. For carnivorous plants, associating with ants may provide defensive benefits in addition to nutritional ones. We examined the effects of increased ant visitation and exclusion of insect prey from pitchers of the hooded pitcher plant Sarracenia minor, which has been hypothesized to be an ant specialist. Visitation by ants was increased by placing PVC pipes in the ground immediately adjacent to 16 of 32 pitcher plants, which created nesting/refuge sites. Insects were excluded from all pitchers of 16 of the plants by occluding the pitchers with cotton. Treatments were applied in a 2 x 2 factorial design in order to isolate the hypothesized defensive benefits from nutritional ones. We recorded visitation by ants, the mean number of ants captured, foliar nitrogen content, plant growth and size, and levels of herbivory by the pitcher plant mining moth Exyra semicrocea. Changes in ant visitation and prey capture significantly affected nitrogen content, plant height, and the number of pitchers per plant. Increased ant visitation independent of prey capture reduced herbivory and pitcher mortality, and increased the number of pitchers per plant. Results from this study show that the hooded pitcher plant derives a double benefit from attracting potential prey that are also capable of providing defense against herbivory.

Adaptive significance and ontogenetic variability of the waxy zone in Nepenthes rafflesiana.

BACKGROUND AND AIMS: The slippery waxy zone in the upper part of pitchers has long been considered the key trapping structure of the Nepenthes carnivorous plants; however, the presence of wax is reported to be variable within and between species of this species-rich genus. This study raises the question of the adaptive significance of the waxy zone and investigates the basis for an ontogenetic cause of its variability and correlation with pitcher shape. METHODS: In Brunei (Borneo) the expression of the waxy zone throughout plant ontogeny was studied in two taxa of the Nepenthes rafflesiana complex, typica and elongata, which differ in pitcher shape and size. We also tested the adaptive significance of this zone by comparing the trapping efficiency and the number of prey captured of wax-bearing and wax-lacking plants. KEY RESULTS: In elongata, the waxy zone is always well expanded and the elongated pitchers change little in form during plant development. Wax efficiently traps experimental ants but the number of captured prey in pitchers is low. In contrast, in typica, the waxy zone is reduced in successively produced pitchers until it is lost at the end of the plant's juvenile stage. The form of pitchers thus changes continuously throughout plant ontogeny, from elongated to ovoid. In typica, the number of captured prey is greater, but the role of wax in trapping is minor compared with that of the digestive liquid, and waxy plants do not show a higher insect retention and prey abundance as compared with non-waxy plants. CONCLUSIONS: The waxy zone is not always a key trapping structure in Nepenthes and can be lost when supplanted by more efficient features. This study points out how pitcher structure is submitted to selection, and that evolutionary changes in developmental mechanisms could play a role in the morphological diversity of Nepenthes.

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.