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

ABSTRACT: 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.

SUBMITTER: Rottloff S

PROVIDER: S-EPMC3170555 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature

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

Rottloff Sandy S Stieber Regina R Maischak Heiko H Turini Florian G FG Heubl Günther G Mithöfer Axel A

Journal of experimental botany 20110601 13

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 bee ...[more]

PMID: 21633084

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Project description:It has been 23 years since the conservation status of highland tropical pitcher plant Nepenthes talangensis was assessed in 2000. A number of existing threats (anthropogenic and environmental) may be increasing the risk of extinction for the species. A better understanding of the ecology and conservation needs of the species is required to manage the wild populations. Specifically, better information related to population distributions, ecological requirements, priority conservation areas, the impact of future climate on suitable habitat, and current population structure is needed to properly assess extinction risks. A better understanding of the requirements of the species in its natural habitat would benefit for successfully securing the species at Botanic Gardens. We have identified 14 new occurrence records of N. talangensis in Mount Talang. Study on the ecological requirement using Random Forest (RF) and Artificial Neural Network (ANN) suggested that elevation, canopy cover, soil pH, and slope are four important variables. The population of N. talangensis was dominated by juvenile and mature (sterile) individuals, we found only a few mature males (7 individuals) and females (4 individuals) in the sampled areas. Our modelling of current conditions predicted that there were 1,076 ha of suitable habitat to very highly suitable habitat in Mount Talang, which is 14.7% of the total area. Those predicted habitats ranged in elevation from 1,740-2,558 m. Suitable habitat in 2100 was predicted to decrease in extent and be at higher elevation in the less extreme climate change scenario (SSP 1-2.6) and extreme climate change scenario (SSP 5-8.5). We projected larger habitat loss in the SSP 5-8.5 compared to the SSP 1-2.6 climate change scenario.. We proposed the category CR B1ab(iii,v), C2a(ii) as the new conservation status of N. talangensis. The status is a higher category of threat compared to the current status of the species (EN C2b, ver 2.3). Nepenthes talangensis seedlings and cuttings established in a Botanic Garden have relatively high survival rate at about 83.4%. Sixty percent of the seeds germinated in growth media successfully grew to become seedlings.

Project description:BackgroundCarnivorous 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.PitcherFluid 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.InquilineDiversity 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).ConclusionsThere 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.

Dataset Information

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

Publications

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

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