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COMMON PART

Project Number22-24-00421

Project titleEndophytic bacteria Herbaspirillum spp. as a basis for the creation of biocontrol preparations increasing productivity of agronomic crops

Project LeadVelichko Natalya

AffiliationInstitute of Biochemistry and Physiology of Plants and Microorganisms Russian Academy of Sciences,

Research area 04 - BIOLOGY AND LIFE SCIENCES, 04-203 - General biochemistry

KeywordsSymbiosis, Herbaspirillum, plant-microbial interactions, plant responses, lipopolysaccharide, O-specific polysaccharide, structure

PROJECT CONTENT

Annotation
The project is aimed at studying the fundamental issues of the interaction of higher plants with soil prokaryotes. The main emphasis will be placed on the systematic study of the peculiarities of the formation of associations with beneficial microflora and plant responses in integrated systems. Traditionally, the attention of researchers is attracted by the search for environmentally friendly and energy-saving approaches for the development of sustainable agriculture. The increasing attention of scientists is focused on the creation of biological products based on microorganisms that stimulate the growth and productivity of plants. Of particular interest among them are endophytic bacteria capable of living mutually inside plant tissues, which allows them, in comparison with other microorganisms, to be less dependent on external environmental factors, exhibit a set of economically useful properties and activate the defense systems of plants, which is adequate and ecologically a safe alternative to agrochemicals. The activation of plant defense systems is mediated by the production of physiologically active compounds produced by microorganisms. One of these elicitors are lipolysaccharides (LPS) of the outer membrane of the cell wall of gram-negative bacteria. At the moment, the phenomenon of mediated regulation of the host immune response has been demonstrated for a wide range of pathogens, while work on the systemic study of the mechanisms of relationships in plant-endophytic systems and the role of LPS of endophytic microorganisms in the induction of defense reactions in plants are presented fragmentarily, which is one of the main reasons. hindering the development of a more effective plant protection system. The genetic potential of the universal bacteria of the genus Herbaspirillum makes it possible to use them not only in the composition of biological products that increase growth and productivity, but also as a model object for studying the mechanisms of mutualistic relationships of higher organisms with endosymbionts. The study of the nature of the distribution of herbapirillum cells in planta, the mechanisms of the manifestation of plant responses in the community with endophytic representatives of these bacteria to the effect of environmental stress factors, as well as the structural features of membrane polysaccharides and their activity against plants is undoubtedly an urgent task. The fundamental basis for solving this problem lies in understanding the principles of the formation and functioning of a specific plant-microbial system. In the course of the project, a characteristic of the process of systemic colonization of different species of cultivated plants will be given, the nature of their response to inoculation by endophytic representatives of Herbaspirillum spp to stress environmental factors will be determined. New, fundamentally important information will be obtained on the structural features of the LPS of poorly studied in this respect gram-negative bacteria of the genus Herbaspirillum. It is known that variations in the structure of LPS can be associated with various molecular mechanisms of activation of the immunological defense of multicellular organisms against infectious agents and their recognition. The most variable part of the LPS molecule is the O-specific polysaccharide (OPS). With the use of modern physicochemical methods of analysis, including MALDI TOF mass spectrometry and NMR spectroscopy, the structure of the repeating unit of the OPS, previously not studied in this regard, the Herbaspirillum strain, will be established. We will study the participation of Herbaspirillum LPS isolated and characterized during the implementation of this project, as well as in our earlier studies, in the induction of a protective response in plants. A comprehensive study of a specific symbiotic system using modern research methods will allow obtaining unique fundamental knowledge and developing biological approaches based on plant-microbial interactions to improve the adaptation of crops to stress factors of biotic and abiotic nature, which is important for establishing biological balance in agrocenoses. The information obtained in the course of the project on the structure and biological activity of LPSs directly involved in the formation of associations of Herbaspirillum with plants can add priority information to information databases for this group of bacteria and expand the fundamental support of the existing ideas about the composition and structure of surface carbohydrate-containing polymers of soil gram-negative bacteria, and will also contribute to the understanding of the processes of formation of associations and draw some conclusions about the participation of LPS of endophytic bacteria in the induction of defense reactions in plants.

Expected results
The implementation of the proposed project will allow obtaining experimental material that will expand the understanding of the principles of the formation and functioning of a specific plant-microbial system. The responses of plants in the community with endophytic representatives of bacteria of the genus Herbaspirillum to stress factors of the environment will be identified, and a qualitative and quantitative assessment of the localization of the studied microorganisms in the tissues of the host plant will be carried out. At the same time, the features of systemic colonization of herbaspirillum inside plant tissues will be described in dynamics in vitro and in vivo. Using recombinant gfp-labeled derivatives, the ability of two promising species of herbaspirillum, previously not studied in this respect, to exist endophytically in the tissues of various cultivated plants, will be determined. An assessment will be made of the influence of the most promising representative of the Herbaspirillum strain investigated during this project on the resistance of wheat plants to pathogenic fungi during the growing season. During the implementation of the project, fundamental knowledge will be obtained about the structures of herbaspirillum lipopolysaccharides and their physiological activity in relation to plants. The structure of the repeating unit of the O-specific polysaccharide of the type strain, previously not studied in this respect, will be established. The study of the influence of structurally different lipopolysaccharides of herbapirillum, which we characterized both during this project and earlier, on the induction of responses of various types of cultivated plants: the synthesis of reactive oxygen species, nitric oxide, salicylic and jasmonic acids will be carried out. The dynamics of changes in the concentration of these compounds in seedlings of cultivated plants under the influence of the selected LPS preparations will be determined. The significance of the results that are planned to be obtained during the work on the project is determined by the need to develop effective and environmentally friendly drugs based on useful forms of prokaryotes to create environmentally safe, highly productive and competitive biological products in adaptive agriculture that can activate the defense systems of plants.

REPORTS