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Carnivorous plants use different morphological features to attract, trap and digest prey, mainly insects. Plants from the genus Nepenthes possess specialized leaves called pitchers that function as pitfall-traps. These pitchers are filled with a digestive fluid that is generated by the plants themselves. In order to digest caught prey in their pitchers, Nepenthes plants produce various hydrolytic enzymes including aspartic proteases, nepenthesins (Nep). Knowledge about the generation and induction of these proteases is limited. Here, by employing a FRET (fluorescent resonance energy transfer)-based technique that uses a synthetic fluorescent substrate an easy and rapid detection of protease activities in the digestive fluids of various Nepenthes species was feasible. Biochemical studies and the heterologously expressed Nep II from Nepenthes mirabilis proved that the proteolytic activity relied on aspartic proteases, however an acid-mediated auto-activation mechanism was necessary. Employing the FRET-based approach, the induction and dynamics of nepenthesin in the digestive pitcher fluid of various Nepenthes plants could be studied directly with insect (Drosophila melanogaster) prey or plant material. Moreover, we observed that proteolytic activity was induced by the phytohormone jasmonic acid but not by salicylic acid suggesting that jasmonate-dependent signaling pathways are involved in plant carnivory.

Mandal and Mukherjee super(1) have discussed about the conservation of pitcher plant, Nepenthes khasiana Hook. f. The plant is endemic to Meghalaya, where it is found in different places of Khasi Hills, Jaintia Hills and Garo Hills super(2). The Khasi people call it 'tiew rakot', which means 'demon flower'; the Jaintia people call it 'kset phare' which means 'fly net with a lid' and the Garo name of this plant is 'memang koksi' which means 'basket of the devil'. The plant is used in the traditional medicine system in Meghalaya. The fresh juice of the pitcher is used in the treatment of asthma, kidney problems, night blindness, skin diseases and leprosy super(2). The juice is also used to treat urinary infection and blockade super(3). The Jaintia people use the juice of closed pitcher as ear drop in ear infections. The pitcher pounded to a paste with water is used in the treatment of cholera. As the traditional medicine system is popular in Meghalaya, a large quantity of this plant is used by the folk healers in the state. In Khasi and Jaintia Hills, about 1878 kg of pitcher is used annually in traditional healing systems super(3). In the Nokrek Biosphere Reserve, Garo Hills, over-exploitation of this plant for medicinal purposes has been identified as one of the causes for decrease in the number of its natural populations super(5). Though there is a report of cultivation of this plan for medicinal use super(3), mostyly natural population are destroyed for the purpose population are destroyed for the pursose. the plant also has ornamental values super(5) and seeding colleted from forests are sold in the market for Rs 20-30 per seeding super(5).

Background and Aims: Carnivorous plants have developed strategies to enable growth in nutrient-poor soils. For the genus Nepenthes, this strategy represents producing pitcher-modified leaves that can trap and digest various prey. These pitchers produce a digestive fluid composed of proteins, including hydrolytic enzymes. The focus of this study was on the identification of these proteins.Methods: In order to better characterize and have an overview of these proteins, digestive fluid was sampled from pitchers at different stages of maturity from five species of Nepenthes (N. mirabilis, N. alata, N. sanguinea, N. bicalcarata and N. albomarginata) that vary in their ecological niches and grew under different conditions. Three complementary approaches based on transcriptomic resources, mass spectrometry and in silico analysis were used.Key Results: This study permitted the identification of 29 proteins excreted in the pitchers. Twenty of these proteins were never reported in Nepenthes previously and included serine carboxypeptidases, α- and β-galactosidases, lipid transfer proteins and esterases/lipases. These 20 proteins display sequence signals allowing their secretion into the pitcher fluid.Conclusions:Nepenthes pitcher plants have evolved an arsenal of enzymes to digest prey caught in their traps. The panel of new proteins identified in this study provides new insights into the digestive process of these carnivorous plants.

This is the first report of silver nanoparticles (AgNPs) synthesis utilizing the pitcher secretion from an insectivorous plant, specifically Nepenthes ventrata , through a microwave assisted green synthesis approach. The successful formation of AgNPs was validated through a comprehensive set of analyses, including UV–Vis spectroscopy, Fourier-transform infrared spectroscopy, transmission electron microscopy, DLS analysis and Zeta potential measurements. In addition gas chromatography-high-resolution mass spectrometry and liquid chromatography-high-resolution mass spectrometry analyses were conducted to examine the components present in the pitcher secretion. These analyses aimed to identify the capping and stabilizing agents in the secretion that facilitate the synthesis and stability of AgNPs. The synthesized AgNPs significantly inhibited biofilm formation by Pseudomonas aeruginosa PAO1, as demonstrated by Crystal Violet staining and fluorescence microscopy. Additionally, these AgNPs showed promising antioxidant properties through a DPPH radical scavenging assay. Furthermore, the anticancer properties of the AgNPs were analyzed using an MTT assay, which measures cell metabolic activity as an indicator of cell viability, proliferation, and cytotoxicity. Collectively, these findings suggest that the biosynthesized AgNPs possess multifaceted biological applications, showcasing their utility as both antimicrobial and antioxidative agents, and highlighting their potential in medical and environmental applications.

The lipid-derived jasmonate phytohormones (JAs) regulate a wide spectrum of physiological processes in plants such as growth, development, tolerance to abiotic stresses, and defence against pathogen infection and insect attack. Recently, a new role for JAs has been revealed in carnivorous plants. In these specialized plants, JAs can induce the formation of digestive cavities and regulate enzyme production in response to different stimuli from caught prey. Appearing to be a new function for JAs in plants, a closer look reveals that the signalling pathways involved resemble known signalling pathways from plant defence mechanisms. Moreover, the digestion-related secretome of carnivorous plants is composed of many pathogenesis-related (PR) proteins and low molecular weight compounds, indicating that the plant carnivory syndrome is related to and has evolved from plant defence mechanisms. This review describes the similarities between defence and carnivory. It further describes how, after recognition of caught insects, JAs enable the carnivorous plants to digest and benefit from the prey. In addition, a causal connection between electrical and jasmonate signalling is discussed.

Limited information exists on the variation and function of pitcher morphological diversity in Nepenthes spp., necessitating further research. Based on a previous study in Mount Pasir Cadas Panjang Ciwidey that identified N. gymnamphora , this research aims to characterize pitcher morphology in Mount Pasir Cadas Panjang Ciwidey. The method used purposive sampling from six locations of N. gymnamphora , pitchers were categorized into upper and lower types based on size. The study revealed distinct differences in shape between upper and lower pitchers. The upper type exhibited a jug-like enlargement at the bottom, narrowing towards the top and enlarging again at the peristome. In contrast, the lower type featured a uniformly enlarged diameter from the bottom to the peristome. Both types shared similar components, differing mainly in wing characteristics the upper type having unhaired and unembossed wings, while the lower type had embossed and haired wings. Pitcher colors included green, patterned, and reddish, with higher elevations correlating with a greater likelihood of reddish or patterned pitchers. This research provides comprehensive data on the morphological and anatomical variations of N. gymnamphora , offering valuable insights for future studies.

Scarcity of essential nutrients has led plants to evolve alternative nutritional strategies, such as myrmecotrophy (ant-waste-derived nutrition) and carnivory (invertebrate predation). The carnivorous plant Nepenthes bicalcarata grows in the Bornean peatswamp forests and is believed to have a mutualistic relationship with its symbiotic ant Camponotus schmitzi. However, the benefits provided by the ant have not been quantified. We tested the hypothesis of a nutritional mutualism, using foliar isotopic and reflectance analyses and by comparing fitness-related traits between ant-inhabited and uninhabited plants. Plants inhabited by C. schmitzi produced more leaves of greater area and nitrogen content than unoccupied plants. The ants were estimated to provide a 200% increase in foliar nitrogen to adult plants. Inhabited plants also produced more and larger pitchers containing higher prey biomass. C. schmitzi-occupied pitchers differed qualitatively in containing C. schmitzi wastes and captured large ants and flying insects. Pitcher abortion rates were lower in inhabited plants partly because of herbivore deterrence as herbivory-aborted buds decreased with ant occupation rate. Lower abortion was also attributed to ant nutritional service. The ants had higher δ(15)N values than any tested prey, and foliar δ(15)N increased with ant occupation rate, confirming their predatory behaviour and demonstrating their direct contribution to the plant-recycled N. We estimated that N. bicalcarata derives on average 42% of its foliar N from C. schmitzi wastes, (76% in highly-occupied plants). According to the Structure Independent Pigment Index, plants without C. schmitzi were nutrient stressed compared to both occupied plants, and pitcher-lacking plants. This attests to the physiological cost of pitcher production and poor nutrient assimilation in the absence of the symbiont. Hence C. schmitzi contributes crucially to the nutrition of N. bicalcarata, via protection of assimilatory organs, enhancement of prey capture, and myrmecotrophy. This combination of carnivory and myrmecotrophy represents an outstanding strategy of nutrient sequestration.

Objectives Nepenthes, sometimes known as tropical pitcher plants or monkey cups, is a carnivorous plant genus that contains more than 160 species. Nepenthes khasiana, India's sole representative of the genus , is a rare and endangered dioecious plant endemic to North-east India. Despite the fact that it is a prominent insectivorous plant in the Nepenthaceae family, genomic resources for the species are limited, making genomic breeding and understanding the genetic basis of botanical carnivory difficult. Herein, we report the complete chloroplast (cp) and mitochondrial (mt) genomes of N. khasiana for the first time. These organelle genomes were assembled as part of a whole-genome sequencing project aimed at gaining deeper insights into their evolutionary relations with genomes of other carnivorous plants. Data description The complete cp genome (156,914 bp) and mt genome (900,031 bp) of N. khasiana are presented here. The cp genome contains two repeat regions and 131 genes (112 unique genes): 86 protein coding genes, 8 rRNA coding genes and 37 tRNA coding genes. The mt genome contains 84 genes (55 unique genes): 50 protein coding genes, 7 rRNA coding genes and 27 tRNA coding genes. The cp and mt genomic data generated will be useful for future molecular characterization and evolutionary research related to botanical carnivory .

The pitcher trap is a striking example of convergent evolution across unrelated carnivorous plant lineages. Convergent traits that have evolved across pitcher plant lineages are essential for trap function, suggesting that key selective pressures are in action. Recent studies have also revealed patterns of divergent evolution in functional pitcher morphology within genera. Adaptations to differences in local prey assemblages may drive such divergence and, ultimately, speciation. Here, we review recent research on convergent and divergent evolution in pitcher plant traps, with a focus on the genus Nepenthes, which we propose as a new model for research into adaptive radiation and speciation.

Nepenthes is a genus comprising carnivorous tropical pitcher plants that have evolved trapping organs at the tip of their leaves for nutrient acquisition from insect trapping. Recent studies have applied proteomics approaches to identify proteins in the pitcher fluids for better understanding the carnivory mechanism, but protein identification is hindered by limited species-specific transcriptomes for Nepenthes . In this study, the proteomics informed by transcriptomics (PIT) approach was utilized to identify and compare proteins in the pitcher fluids of Nepenthes ampullaria , Nepenthes rafflesiana , and their hybrid Nepenthes × hookeriana through PacBio isoform sequencing (Iso-Seq) and liquid chromatography-mass spectrometry (LC-MS) proteomic profiling. We generated full-length transcriptomes from all three species of 80,791 consensus isoforms with an average length of 1,692 bp as a reference for protein identification. The comparative analysis found that transcripts and proteins identified in the hybrid N. × hookeriana were more resembling N. rafflesiana , both of which are insectivorous compared with omnivorous N. ampullaria that can derive nutrients from leaf litters. Previously reported hydrolytic proteins were detected, including proteases, glucanases, chitinases, phosphatases, nucleases, peroxidases, lipid transfer protein, thaumatin-like protein, pathogenesis-related protein, and disease resistance proteins. Many new proteins with diverse predicted functions were also identified, such as amylase, invertase, catalase, kinases, ligases, synthases, esterases, transferases, transporters, and transcription factors. Despite the discovery of a few unique enzymes in N. ampullaria , we found no strong evidence of adaptive evolution to produce endogenous enzymes for the breakdown of leaf litter. A more complete picture of digestive fluid protein composition in this study provides important insights on the molecular physiology of pitchers and carnivory mechanism of Nepenthes species with distinct dietary habits.