Badania strukturalne i funkcjonalne roślinnych enzymów metabolizmu argininy jako molekularny cel w projektowaniu nowych środków ochrony roślin.
Structural and functional studies of the plant enzymes of arginine metabolism as molecular targets of novel plant protection products.
Typ projektu: naukowo-badawczy
Słowa kluczowe: Biosynteza argininy asymilacja azotu odpowiedź na stress biologia strukturalna mechanizmy obronne roślin herbicydy
Słowa kluczowe (angielski): Arginine biosynthesis nitrogen assimilation stress response structural biology defense mechanisms in plants herbicides
Członkowie konsorcjum: Projekt nie był realizowany w ramach konsorcjum
Okres realizacji projektu: 3.10.2022 - 2.10.2025
Instytucja finansująca: Narodowe Centrum Nauki
Nazwa programu: Sonata
Kierownik projektu: Bartosz Sekuła
Wartość dofinansowania: 1 781 200,00 PLN
Wartość projektu: 1 781 200,00 PLN
Plants have developed advanced defensive mechanisms to grow under stress-inducing conditions, like drought, nutrient shortage, extreme temperatures, or invasion of pest and pathogens. Arginine is a central part of plant development and defensive response mechanisms; it serves not only as a building block of proteins but also as an effective storage form of organic nitrogen, and a precursor of many defensive metabolites, like polyamines, proline, or glutamate. After nitrogen uptake, plants synthesize ornithine through the cyclic or linear pathways. Then, as a part of the urea cycle, arginine is produced. Arginine reserves are mobilized to synthesize and accumulate protective catabolites under stress conditions. Although the general organization of plant arginine metabolism seems to be similar to other eucaryotes and procaryotes, the specific function of the pathway and each enzyme significantly differentiate plants from species of other domains of life. In general, animals use urea cycle to excrete excess of nitrogen, while plants recycle it, because it the most limiting essential element for plant development. The studies on plant arginine metabolism have seen a tremendous development in recent years, but the research has been limited mainly to genetic approach, with marginal studies at the protein level. The project is aimed to explain one of the most essential plant metabolic pathways that is far from full understanding. It tackles structural and functional characterization of the selected plant enzymes involved in arginine production and conversion. The studies will be conducted with structural (X-ray crystallography, cryo-electron microscopy), biochemical (enzymatic assays), and biophysical (affinity studies) methodology. The enzymes will be thoroughly investigated, their reaction mechanisms and substrate specificity will be characterized. Additionally, the determined molecular structures with various ligands (substrates, products, cofactors, etc.) will be used to pinpoint key structural features that drive arginine biosynthesis and conversion in plants. This will become a basis for rational design of site-directed functional mutants and investigations of specific activators/inhibitors of the studied enzymes that modulate the rate of their action. It leads to the potential important practical applications in agriculture. It was shown that enzymes from arginine metabolism are blocked by natural toxins produced by pathogenic bacteria. Specific inhibition of the process of arginine production or mobilization, similarly to natural phytotoxins, can be promising mode of action of novel herbicides. Such solutions are essential in the light of approaching legislative withdrawal of many plant protection products and alarming rising resistance to currently used substances observed in weeds, pests, and pathogens. On the other hand, crops absorb only a fraction of nitrogen from applied fertilizers (~35% in the most optimal conditions). Activators or functional mutants that facilitate arginine production and turnover may be used to increase the efficacy of nitrogen uptake; they may also provide additional protection against various stress conditions through the accumulation of defensive compounds. An example of practical use of this feature is the action of overexpressed arginase in tomatoes that inhibited growth of some pest larvae through depletion of arginine. Agricultural food production struggle with many environmental (rapid soil degradation, increasing pollution), health (harmful effect of pesticides and herbicides, e.g., potential carcinogenicity of glyphosate), and economical (nitrogen is the most expensive element of fertilizers) challenges and without novel safe solutions future sustainability of food production may be compromised. The results of the project will bring not only cognitive value (novel structures and deeper understanding of the primary metabolism in plants), they will also provide a groundwork for the future development of novel and safer solutions for agriculture.
Plants have developed advanced defensive mechanisms to grow under stress-inducing conditions, like drought, nutrient shortage, extreme temperatures, or invasion of pest and pathogens. Arginine is a central part of plant development and defensive response mechanisms; it serves not only as a building block of proteins but also as an effective storage form of organic nitrogen, and a precursor of many defensive metabolites, like polyamines, proline, or glutamate. After nitrogen uptake, plants synthesize ornithine through the cyclic or linear pathways. Then, as a part of the urea cycle, arginine is produced. Arginine reserves are mobilized to synthesize and accumulate protective catabolites under stress conditions. Although the general organization of plant arginine metabolism seems to be similar to other eucaryotes and procaryotes, the specific function of the pathway and each enzyme significantly differentiate plants from species of other domains of life. In general, animals use urea cycle to excrete excess of nitrogen, while plants recycle it, because it the most limiting essential element for plant development. The studies on plant arginine metabolism have seen a tremendous development in recent years, but the research has been limited mainly to genetic approach, with marginal studies at the protein level. The project is aimed to explain one of the most essential plant metabolic pathways that is far from full understanding. It tackles structural and functional characterization of the selected plant enzymes involved in arginine production and conversion. The studies will be conducted with structural (X-ray crystallography, cryo-electron microscopy), biochemical (enzymatic assays), and biophysical (affinity studies) methodology. The enzymes will be thoroughly investigated, their reaction mechanisms and substrate specificity will be characterized. Additionally, the determined molecular structures with various ligands (substrates, products, cofactors, etc.) will be used to pinpoint key structural features that drive arginine biosynthesis and conversion in plants. This will become a basis for rational design of site-directed functional mutants and investigations of specific activators/inhibitors of the studied enzymes that modulate the rate of their action. It leads to the potential important practical applications in agriculture. It was shown that enzymes from arginine metabolism are blocked by natural toxins produced by pathogenic bacteria. Specific inhibition of the process of arginine production or mobilization, similarly to natural phytotoxins, can be promising mode of action of novel herbicides. Such solutions are essential in the light of approaching legislative withdrawal of many plant protection products and alarming rising resistance to currently used substances observed in weeds, pests, and pathogens. On the other hand, crops absorb only a fraction of nitrogen from applied fertilizers (~35% in the most optimal conditions). Activators or functional mutants that facilitate arginine production and turnover may be used to increase the efficacy of nitrogen uptake; they may also provide additional protection against various stress conditions through the accumulation of defensive compounds. An example of practical use of this feature is the action of overexpressed arginase in tomatoes that inhibited growth of some pest larvae through depletion of arginine. Agricultural food production struggle with many environmental (rapid soil degradation, increasing pollution), health (harmful effect of pesticides and herbicides, e.g., potential carcinogenicity of glyphosate), and economical (nitrogen is the most expensive element of fertilizers) challenges and without novel safe solutions future sustainability of food production may be compromised. The results of the project will bring not only cognitive value (novel structures and deeper understanding of the primary metabolism in plants), they will also provide a groundwork for the future development of novel and safer solutions for agriculture.
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