Laboratory of Microbial Ecology

Head Kozyrovska Natalia, PhD. Tel.: +38044 5265596 Fax: +38044 5260759 E-mail: kozyr@imbg.org.ua

Research Background:

Metagenomic and Biotechnological Aspects in a Study of the Endophytic Bacteria Role in Optimization Plant Nutrition and Protection

Changes in resident endophytic bacterial community structure in model system ‘in vitro potato (Solanum tuberosum L.) - plant-associated bacteria’ under a process of microclonal propagation were detected: some bacteria fall into an unculturable state, and they can be detected with molecular tools only. In this way, a number of culturable bacteria decreases. There is correlation between decline of a number of operational taxonomic units and a vulnerability of vitroplants ex vitro. We were able to show that bacteria which resided in plant tissue in unculturable state became culturable after inoculation with a biological agent. Priming the in vitro-grown potato with non-pathogenic bacteria results in increase of the diversity of culturable endophytes, and so far in inducing plant systemic resistance and improved plantlets survival ex vitro.

А	Б
Fig. In situ hybridization (А) and fluorescence іn situ hybridization (B) of in vitro potato root tissue inoculated with Methylobacterium radiotolerans IMBG290 with digoxigenin labeled specific probe. Р – parenchyma; X/V – xylem vessel. Scale bar = 50 μm.

Modelling Plant Growing under Assistance of Microbial Consortia in Extraterrestrial Green Houses

A concept of growing pioneer plants and protosoil formation in Lunar green house under assistance of a rationally assembled microbial community has been elaborated in our lab under the aegis of National Space Agency and European Space Agency. Together with researchers of National Taras Shevchenko University (selection of analogs of lunar regolith; rock elemental composition testing; biomobilizing nutritional elements and their accumulation in plant biomass), National M. Gryshko Botanical Garden (a plant selection), Іnstitute of F. Ovcharenko Biocolloidal Chemistry of NASU (physical and chemical interaction of bacteria with rocks), Institute of Physics of NASU (modeling impact of a low-dose radiation on bacteria in a confined system) we have got results in a support of this concept.

Fig. Marigold (Tagetes patula L.) grown on anorthosite of Turchynka type inside the ‘PlantKord’, a locker-based facility for plant cultivating.

Current research activities

• Plant-microbe interactions: mechanisms of plant protection against bacterial and viral diseases assisted by plant-associated bacteria and bioactive compounds; optimization of plant mineral nutrition and protection under growth-limiting conditions.
• Endophytic bacterial communities in model systems. Metagenome of endophytes.

Collaboration

• Kyiv T. Schevchenko University
• National M. Gryshko Botanical Garden
• Institute of F. Ovchrenko Biocolloidal chemistry of NANU
• Institute Physics of NANU
• Potato Institute of UAAS
• ESA Research and Scientific Support Department, ESTEC/SCI-PD, Noordwijk, The Netherlands, Dr. B.H. Foing
• Oulu University, Finland, Dr. A.M. Pirtilla

Award

The World Intellectual Property Organization award for the best invention (2003)

Selected publications:

1. Podolich O., Laschevskyy V., Ovcharenko L., Kozyrovska N., Pirttilä A. M. Methylobacterium sp. resides in unculturable state in potato tissues in vitro and becomes culturable after induction by Pseudomonas fluorescens IMGB163. J. Appl. Microbiol., 2008 (accepted) .
2. Ovcharenko L.P., Kozyrovska N.O. Metagenomic analysis of microorganisms from the environment. Kyiv, 2008, Sprint-Print, 252 p. (in Ukr.).
3. Lar O., Kozyrovska N. Labelling gram-negative bacteria with the lux- and gusA reporter genes. In: Prospects and applications for plant-associative bacteria, eds S. Sovari, A.M. Pirttilä, 2008, p.56-61.
4. Kovtunovych G., Kovalchuk M., Lar O., Kamalova S., Kozyrovska N. Use of the gusA- and lux- reporter genes in monitoring plant-bacteria interactions. In: Prospects and applications for plant-associative bacteria, eds S. Sovari, A.M. Pirttilä, 2008, p. 214-218.
5. Podolich O. V. Ardanov P. E., Voznyuk T. M., Kovalchuk M. V., Danylchenko O. V., Laschevskyi V. V., Lyastchenko S. A., Kozyrovska N. O. Endophytic bacteria from potato in vitro activated by exogenic non-pathogenic bacteria. Biopolymery i кlityna, 2007, 23 (1): 21-28.
6. Kozyrovska N.O., Korniichuk O.S, Voznyuk T.M., Lytvynenko T.L., Kononuchenko O., Zaetz I., Rogutskyy I.S., Mytrokhyn O.V., Mashkovska S.P., Foing B.H., Kordyum V.A Growing pioneer plants for a lunar base. Adv. Space Res., 2006, 37: 93-99.
7. Lytvynenko T., Zaetz I., Voznyuk T., Kovalchuk M., Rogutskyy I., Mytrokhyn O., Lukashov D., Estrela-Lopis V., Borodinova T., Mashkovska S., Foing B., Kozyrovska N. A rationally assembled microbial community for growing Tagetes patula L. in a lunar greenhouse. Research in Microbiol., 2006, 157: 87-92.
8. Zaets I., Voznyuk T., Kovalchuk M., Rogutskyy I., Mytrokhyn O., Lukashov D., Mashkovska S., Foing B., Kozyrovska N. Optimization of plant mineral nutrition under growth-limiting conditions a lunar greenhouse. Kosmichna Nauka i Technologiya, 2006, 12 (4) : 1-8.
9. Kozyrovska N., V. Negrutska, M. Kovalchuk, T. Voznyuk. Paenibacillus sp., a promising candidate for development of a novel technology of plant inoculant production. Biopolymery i klityna, 2005, 21 (4): 312-319.
10. Kozyrovska, N.O., Korniichuk, O.S., Voznyuk, T.M., et al. Microbial community in a precursory scenario of growing Tagetes patula in a lunar greenhouse// Kosmichna Nauka i Technologiya.- 2004, 10 (5-6): 221-225.
11. Kovtunovych G., Lytvynenko T., Negrutska V., Lar O., Brisse S., Kozyrovska N. Identification of Klebsiella oxytoca using a specific PCR assay targeting the polygalacturonase pehX gene. Research in Microbiol., 2003, 154: 587-592.
12. Kovtunovych G. L., Lytvynenko T., Negrutska V. V., Lar O. V., Koltukova N. V., Kozyrovska N. O. A PCR-mediated method for discrimination of Klebsiella oxytoca between closely related bacteria in environmental and clinical specimens. Biopolymery i klityna, 2003, 19 (6): 520-525.
13. Kovalchuk M. V., Negrutska V. V., Kovtunovych G. L., Lar O. V., Korniichuk O. S., Rogutskyi I. S., Alpatov A. P., Kozyrovska N. O., Kordyum V. A. Modeling the pNARSLux transfer in the wheat rhizosphere under simulated microgravity// Kosmichna nauka i technologiya (Suppl.).- 2003, 9 (2): 10-14.
14. Dong Y., Chelius M., Brisse S., Kozyrovska N., Kovtunovych G., Podschun R., Triplett E.W. Comparisons between two Klebsiella: the plant endophyte K. pneumoniae 342 and a clinical isolate, K. pneumoniae MGH78578. Symbiosis, 2003, 35: 247-259.
15. Lar O.V., Kovtunovych G.L., Kozyrovska N.O. Cloning and analysis of the gene encoding pectate lyase, the Klebsiella oxytoca VN13 pelX. Biopolymery i klityna, 2002, 18 (5): 417- 422 (Ukr.).
16. Kovtunovych G.L., Lar O.V., Kozyrovska N.O. Cloning and structural analysis of the pehX gene of Klebsiella oxytoca VN13. Biopolymery i klityna, 2000, 16 (5): 356-363 (Ukr.).
17. Kovtunovych G., Lar O., Kamalova S., Kordyum V., Kleiner D., Kozyrovska N. Correlation between pectate lyase activity and ability of diazotrophic Klebsiella oxytoca VN 13 to penetrate into plant tissues. Plant Soil, 1999, 215: 1-6.
18. Kovtunovych G., Lar O., Kordyum V., Kleiner D., Kozyrovska N. Enhancing the internal plant colonization rate with endophytic nitrogen-fixing bacteria. Biopolymery i klityna, 1999, 15 (4): 300-306.
19. Kozyrovska N., Kovtunovych G., Gromosova O., Kuharchuk P., Kordyum V. Novel inoculants for an environmentally-friendly crop production. Resources, Conserv. and Recycling, 1996, 18:79-85.