Home LABORATORY OF BIOCHEMISTRY AND PLANT IMMUNITY


[Head of Laboratory ] [Structure of Laboratory] [The areas of interests] [Main achievments] [Connactions] [Grants]



Head of Laboratory:

    academician I.A.TARCHEVSKY,
    tel.:   +7(495)-958-55-26
    e-mail: tarchevsky@inbi.ras.ru
                tarchevsky@mail.knc.ru

back

Structure of Laboratory:
    Group "Induced Resistance of Plants"


    Head: Ozeretskovskaya Olga Leonidovna, D. Biol. Sci., professor, known specialist in the area of plant defence from disease and stresses.
      Group of Regulation of Plant Dormancy and Resistance to Disease

    Head: Nataliya P. Korableva, Ph. D., D. Biol. Sci., professor
    tel.:  +7(495)-954-4097
    e-mail:
    ozeretskovskaya AT inbi.ras.ru
      tel.:  +7(495)-954-4097
    e-mail:
    korableva AT inbi.ras.ru

    The laboratory consists of 4 doctors of biological sciences, 5 candidates of biological sciences, 6 workers without scientific degrees.
back

The areas of interests:
         The investigations are applied to the study of mechanisms of phytoimmunity, which was the basis for the plant resistance against biotic and abiotic stresses, the signal systems, determining the induction or inhibition of plant immune responses and also of the processes of wound reparation. The principle of the induction of systemic prolonged plant resistance with biogenic elicitors, evolved early by the researchers of laboratory, allows to increase of immune potential of plants and has made possible to increase the crop, the quality of the agricultural production and the more long storage period.
         There is the screening of elicitors with systemic prolonged action is realized for increasing plant resistance against stresses and diseases. The components of pathogens and plants (polyenoic fat acids, fragments of chitin and chitosan. β-1,3-glucans, xyloglucans, and also composition of elicitors and signal molecules and their fragments are used as elicitors. The interplay of structure and function of immunomodulators, determinating the defense mechanisms of plants against biotic and abiotic stresses are investigated.
         The investigation of process of plant tissues wound reparation, which is the one of more ancient defense reactions, arrests our attention. The rate and intensity of wound reparation are investigated on a basis of the estimation of forming of the second meristem - phellogen and defense properties of periderm, and this is essential for safety the crop of agricultural production.
         Fundamental studies on biochemical and molecular mechanisms of the regulation of plant growth and resistance to biotic (plant pathogens) and abiotic (unfavorable temperature, water deficiency at al.) stresses are the major problems to be investigated. The investigations are based on concept about interrelations of ontogenetic stage and resistance in plants. As had been shown by long-standing investigations, the development of immune response is related to the length of deep dormancy in potato tuber of different variety that is determined genetically. The capacity of plant tissues to enhanced synthesis of therpenoids and phenolics (the hormone of dormancy – abscisic acid, constitutive and induced antibiotics) is one of the important constituents of defensive response.
         A study of the signal systems of plant cells and their role in the development of resistance is the topical direction in the field of plant dormancy and immunity regulation. Studies of molecular mechanisms of the action of signal molecules (ethylene, abscisic and jasmonic acids, brassinosteroids at al.) includes the evaluation of functional activity of plasma membrane enzymes (of H+ATPase, Ca+ATPase), modification of cell ultrastructure, changes in stress phytohormone biosynthesis, variations of the role of polygalacturonase inhibiting protein (PGIP) in the resistance. The investigation of PGIP is of importance because it increases plant resistance to diseases by inhibition of the activity of polygalacturonase – main pathogenic factor of pathogen, and besides, is involved in the regulation of processes of plant cell growth and cell wall elongation at different ontogenetic stages.
         The results of the investigations are the basis for further study of mechanisms of the interaction between different systems of signal transduction into plant cell, biochemical processes in plant related to the regulation of growth, dormancy, aging, and immunity. The using for the experiments of transgenic plants with inserted genes of taumatin, defensin et al. that are involved in the regulation of resistance allows to reveal post transformational changes in the reaction of plants on the effect of signal molecules and physiologically active substances.
         The screening of the inducers of dormancy and resistance for development of ecologically harmless methods to increase the resistance of potato, vegetables and fruits during cultivation and storage of agricultural products.
back

Main achievments:
          The increase of expression of plant defense genes, the rate of spreading of defense effect and duration its safety by composition preparations of elicitors and some mediators of signal systems or their fragments were first established.
          The screening of chitosan derivatives allowed to reveal the new elicitor – N-(2-hydroxy-3methoxybenzyl-)-N-piridoxchitosan, which stimulated the resistance of potato against Phytophthora infestans and reparated the wounds of potato tubers more essentially than initial elicitor. There are the concentrations, doses, time and modes of using this elicitor in the recent biotechnology of increase production of agricultural variety were optimized.
          It was established that the content of salicylic acid in potato tissues raised in response to pathogen or treatment by elicitors in the result the activation of enzymes synthesis of salicylic acid or the hydrolysis its conjugated forms. Salicylic acid is irreversibly inhibited the enzymes of antioxydant complex as were shown on two pathosystems – potato – P. infestans and tomato- gall nematodes. The level of hydrogen peroxide increases and “oxidative burst”, which is one of the important mechanisms of plant systemic prolonged resistance, is generated after inhibiting of enzymes of antioxidant complex by salicylic acid.
          It was shown the participation of protein-inhibitors of proteinase at defense reactions potato tubers against phytopathogens and wound reparation
          It was established that wound reparation provided by systemic long action.
          It was established that nanoparticles of chitosan and also nanocrystalic metalic powder and selen stimulated defense effects of potato tubers and its shooting.
          The immunosuppressor –laminarin, possessing nonspecific activity to inhibit immune responses of plants , and the specific suppressor –lowmolecular β-1,3-β- 1,6-glucan, inhibiting plant tissues immune responses only at the susceptible relationship of potato and P. infestans , were detected at the investigation of mechanisms of susceptibility of potato to P. infestans. It was established, that both types of suppressors possess antielicitor action and prevented the induction of potato resistance, evoked by elicitors or uncompatible race of pathogen. It was shown that susceptibility potato to P. infestans, which induced by immunosupressors was one from reasons of bacterial diseases accompanying phytophthorosis. It was established that the present of immunosuppressor in work solutions used for plant immunization was able to inhibit immune responses of potato and tomato, inducing the susceptibility to disease instead of resistance.
          Study of plant dormancy and resistance is based on idea about interaction of biochemical mechanisms controlling these two integral functions of plant organism. It was shown that break of dormancy in potato tuber apices is connected to action of phytohormones regulating morphogenesis, RNA and protein synthesis, function of plasma membrane and the most important cell signal systems. In the induction of deep dormancy and resistance the crucial role belongs to two stressful phytohormones – ethylene and abscisic acid.
          Electron-mycroscope and morphometry investigations demonstrated that cell enlargement is the leading process in initial stages of potato tuber sprouting. The cells of central and stem meristem in potato tuber apices were the target for the action of synthetic growth regulator epibrassinolide. Effect of antioxidant ambiol on dimentions of mitochondria and plastides and development of some intraplastid membrane structure in the cells of potato tuber apices of initial plant differed from these of plants transformed by defensine gene. The influence of ambiol on the formation of stems, leaves and roots of initial potato plants differed from those of transformed ones.
          The binding of phytohormones gibberellic acid (GA) and abscisic acid ABA) to plasma membrane vesicles (PMV) prepared from potato tuber cells was ascertained. The binding of GA and ABA to PMV proteins resulted in conformation rearrangement of some part of protein molecule affecting physico-chemical properties and structure of local areas of plasma membrane surface. Proteinkinase C activity was found in PMV. The activity increased as a result of potato tuber treatment with GA. Phytohormones influenced also the activity of plasma membrane-bound H+ATPase of potato tuber cells.
          It was revealed that the activity of plasma membrane H+ATPase and the enzyme sensitivity to physiologically active substances changes as a result of introduction in potato plants genes of biologically active peptides defensine and thaumatin, which increase the plant resistance to phytophathogenes. The nature of the changes depended on the stage of ontogenesis (deep dormancy, end of dormancy, sprouting).
          Electron-mycroscope study found some alterations in cell ultrastucture induced by penetration of Phythophthora pathogen in potato cell. Investigation of biochemical processes on the contact surface of pathogen fungus and host plant discovered exstracellular metabolites of Phytophthora infestans with high molecular weight. The metabolites bound to PMV from potato tuber cells competing to phytohormones for membrane binding sites.
          The pectolytic enzyme complex of phytophathogen fungus Verticillium dahliae, Rhizoctonia solani, P. infestans was studied. A role of polygalacturonase inhibiting protein (PIPG) in biochemical reactions arised from the contact of phytophathogen fungus with plant tissues is examined. The dynamics of PIPG activity during potato plant vegetation, potato tuber storage and apple-tree fruits maturation and storage was observed. For a first time the sinusoidal alteration in PIPG activity during plant ontogenesis was found.
          It was shown that polygalacturonase inhibiting protein (PGIP) take a part in the regulation of frut ripening and resistance to infections. The PGIP activity varied in apple and banana fruits depending on the stage of ripening and is related to the intencity of etylene production. The inhibition of etylene production by the action of aminoacetic acid or its stimulation by the action of 2-chlorethylphosphonic acid caused the decrease or the increase of PGIP activity.
          For a first time nucleotide and amino acid successions of PIPG from Solanum tuberosum and Solanum brevidens were determined and deposited in gene bank NCBI (in the common work with Speransky A.S., Bogatcheva A.M., Schevelev A.B., Krinitsina A.A.). The cloning and sequening of PGIP genes from S.tuberosum and S. brevidens allow to estimate the multiplicity and the degree of polymorphism of PGIP geens in the genoms of investigated species and appreciate its role in the plant defensive reactions.
          The important role of phytohormone ethylene in the regulation of suppy fruit maturation was established. Some changes in the balance of precursors of ethylene synthesis in methyonine cycle – aminocyclopropanecarbonic acid (ACC) and malonyl-ACC as well in ACC-synthase activity was found in the course of apple fruit maturation and functioning of two NADHP-generating systems: malic enzyme and glucose-6-phosphate dehydrogenase in the fruit tissues. The model of scrining of dormancy and resistance inducers was developed. This model is used for search of physiologically active substances playing a role of both growth regulators and inducers of resistance to pathogens and other unfavorable factors.
          The parameters of application of complex prepatation ethacide for the treatment of fleshy fruits and hevea trees. The result of investigations provides the basis for biotechnology of the application of ethacide for the reduction of product losses during the storage of apple and banana fruits by the controlling the rate of ripening and the increase of immunity. The biotechnology is applicable also on the hevea plantation for the incrase of latex excretion in the conditions of Vietnam tropics.
          The parameters of the application of antioxidant astaxantine would be used for the development of biotechnology preventing aging and apoptosis within the tissues of fleshy storage plant organs during after harvesting. In common with “Bioengineering” centrum of RAS new results were obtained concerning transgenic plants with the embeded genes of defensine and taumatine that increase the immunity of potato plants. The possibility was shown of application of bioregulators for the positive correction of growth and development of transgenic potato plants and for the increase the reproductive capability of modern pea morphogenotypes.Tese data are important for further perspective work.
back

Interinstitutes and international scientific connections:
    Center “Bioengineering” RAS, Moscow (Skryabin G.K., acad.)
    Timiryasev Institute of Physiology of Plants RAS,
    Institute of Defence Plants Russian Academy of Agriculture.
    Moscow University ( Department of biology, mycology,algology),
    Russian Institute of Phytopathology,
    N.M. Emmanuel Institute of biochemical physics RAN, Moscow (Smirnov L.A., prof.)
    M.E. Arbuzov Institute of organic and physic chemistry KazSC RAN, Kazan (Reznik A.I., d.ch.sci.)
    Ogarev State University, biological faculty, Orel (Puzina T.I., d.b.sci.)
    Horticultural Institute RAAS, Mitchurinsk (Gudkovskii V.A., acad.)
    Siberian Institute of Plant Physiology and Biochemistry RAS (Romanenko A.S., d.boil.sci.)
    Institute of tropical biology, Saigon, Vietnam (Nguen Tien Thang, D.Ph., Ngo Ke Syong, D.Ph.)
    Institute of experimental botany, Prague, Czechia (Mahachkova I., D.Ph.) Lorch Institute of potato,
back
Grants:
    1. Russian Foundation for Basic Research Grant ¹ 98-03-32144
    «Electrochemical properties of monoamino oxidases in reactions with artificial acceptors of electrons», 1998-2000.
    2. INCO Copernicus ¹ ICA2-CT-2000-10050
    «Pulp biodegradation using lignolytic enzyme reactions», 2000-2003.
    3. FNTP of Russian Ministry of Science and Technology «Biocatalytic technologies» ¹ 43.073.1.1.2505
    «Pulp biodegradation using laccase-mediator systems», 2000-2003.
    4. Russian Foundation for Basic Research Grant ¹ 02-04-48885 «Development and study of mechanism of environmentally friendly synthesis of water-soluble conducting polyaniline by laccases», 2002-2004.
    5. Russian Foundation for Basic Research Grant ¹ 03-04-48937
    «Free radical reactions in the laccase-mediator system under enzymatic degradation of xenobiotics», 2003-2005.
    6. FNTP of Russian Ministry of Science and Technology ¹ 02.467.11.3004
    «Development of industrial enzyme-based preparations for pulp and paper industry, textile industry, and bonding agents, and biologically-compatible plastics» from March 30, 2005.
    7. INTAS ¹ 03-51-6278
    «Novel biosensors and analysis kits based on genetically engineered biomolecules for formaldehyde assay», 2004-2006.
    8. Grants- RFFI –(2000-2002, N 00.04.49109; 2006-2008, N 06.04.48924);
    9. Phytobiotechnology (2002-2004)
    10. RFFI - ¹10-04-00792a (2010-2012)
    "The investigation of immunological mechanisms of relationships between parasitic nematodes and plants: the role of elicitors and signal molecules"
    11. RFFI - ¹ 07-04-90000, 2007 – 2008
    ”Molecular mechanisms of ripening and disease resistans in the tropical plant fruits under the action of synthetic growth regulators”.
    12. RFFI ophi - 07-03-12046. ¹ 4 / 2007
    13. MES. 2007-2-1.2-00-02. ¹ 02.11. 512.2158. 2007-2010
back


[Main Page]

email to INBI
Last review: 19, October, 2010
© A.N.Bach Institute of Biochemistry of the R.A.S., 2001-2010