DEVELOPING LEAF TRAPS OF CARNIVOROUS SUNDEWS: SOURCE OF UNIQUE HYDROLASES WITH A HIGH POTENTIAL IN BIOTECHNOLOGY
APVV-23-0448 (1.9.2024-30.6.2028)
Principal Investigator: Jana Libantová
Carnivory is a special case of a functional adaptation to allow growth in nutrient-poor soils. Sundews, carnivorous plants of the Drosera genus employ modified leaves (traps) that serve multiple functions, including prey capture, digestion and nutrient absorption, and photosynthesis. The structural and morphological changes in the leaves begin during the early stages of development. Within this project’s scope, we aim to investigate the developmental stages of sundew leaves, when the functional trapping organs form and hydrolytic enzymes are produced, using transcriptomics and proteomics. The screening data will be used to generate a list of genes or proteins relevant to development or involved in prey digestion. Subsequently, we will isolate candidate genes related to digestion and assess their organ-specific expression profile, thereby contributing to a better understanding of the energy cost of trap functionality. Based on previous research, in which we established the production of a sufficient amount of purified recombinant proteins – chitinase and β-1,3-glucanase from Drosera binata and Drosera rotundifolia in an Escherichia coli expression system, we will biophysically, biochemically, and structurally characterize these enzymes. We will also identify ligands that modulate their activity and stability. In addition, we will evaluate the potential of hydrolytic enzymes from the genus Drosera for biotechnological applications by testing whether the combination of these enzymes affects the growth of selected phytopathogens.
STUDY OF HYDROLASES INVOLVED IN DIGESTION IN CARNIVOROUS PLANTS OF THE GENUS DROSERA
VEGA 2/0021/24 (1.1.2024-31.12.2027)
Principal Investigator: Jana Libantová
Carnivorous plants, including species of the genus Drosera, use a mixture of enzymes secreted by stalked glands of the leaf cuticle – the tentacles – to digest their prey. The proposed project involves the analysis of leaf proteins, including those involved in digestion, by a proteomic approach. Based on this, we will select a new hydrolase for more detailed study. Available orthologous gene sequences of related plant species will be used for primer design, and then the gene fragment and the complete gene will be isolated by PCR. The expression profile of the gene under investigation in the individual organs of Drosera plants will also be analysed. The project also includes the introduction of a eukaryotic expression system for the production and characterisation of proteins with post-translational modifications. Based on the results of previous research, some additional enzyme characteristics of purified chitinases and glucanases from carnivorous plants with potential for biotechnology will be investigated.
MOLECULAR CLONING AND BIOCHEMICAL CHARACTERISATION OF A PECULIAR PROTEASE FROM CARNIVOROUS SUNDEW
09I03-03-V04-00573 (1.7.2024-30.6.2026)
Principal Investigator: Monika Danchenko
Carnivorous plants evolved leaves with the specialised function of capturing and digesting insect prey and absorbing the desired nutrients while growing on deficient soils. Among them, sundews secrete a rich mixture of digestive enzymes by the stalked glands of the leaf skin—tentacles. To date, the genomes of individual species, including genes involved in digestion, have been poorly explored. Therefore, the fellowship focuses on the analysis of an in silico predicted hydrolytic enzyme active in digestion processes. Firstly, the project aims to isolate a selected gene coding a putative sundew protease and evaluate its expression profile in plant organs. Secondly, the project will employ a eukaryotic expression system for the synthesis of the recombinant protease and, finally, characterise the biochemical and functional properties of the purified protein using state-of-the-art methodologies. The implementation of the fellowship will enhance career prospects of the Researcher, advance the specific scientific field of carnivorous plants biology, and positively influence society by educating students and inspiring nature protection activists.