Biopolym. Cell. 1999; 15(4):300-305.
http://dx.doi.org/10.7124/bc.00052A
Cell Biology
Enhancing the internal plant colonization rate with endophytic nitrogen-fixing bacteria
1Kovtunovych G. L., 1Lar O. V., 1Kordyum V. A., 2Kleiner D., 1Kozyrovska N. O.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Department of microbiology, Unversity of Bayreuth
    95440 Bayreuth, Germany

Abstract

Several diazotrophic strains of Klebsiella oxytoca and K. terrigena that colonize the plant-host interior were able to produce the plant cell wall depolymerising enzyme pectate lyase (Pel). The activity of the K. oxytoca enzyme was weaker than that of phytopathogenic bacteria, and it was located mainly inside the cells. A small fraction of the cells (10–6 to 10–5) in populations grown in nonselective media was able to grow in a selective medium with polygalactorunate (PC) as sole carbon source. After passage through selective medium cells were converted to the Pet+ -phenotype, and total Pet-activity in population of K. oxytoca increased. The increased level of Pelactivity of K. oxytoca and K. terrigena correlated with a 10-fold higher rate of internal colonization of wheat roots. Cultures of K. oxytoca VN13 grown in selective medium with PG also showed increased stimulation of wheat growth. Seedlings inoculated with such cultures exhibited better development resulting in higher biomass.
Full text: (PDF, in English)

References

[1] Kordyum V. A., Kozyrovskaya N. A., Gunkovskaya N. V. The theoretical basis and experimental construction of new-type nitrogen fixers able to grow and fix the atmospheric nitrogen in nonlegumes. Biopolym. Cell. 1986; 2(5):227-239
[2] Nguyen T. N. H., Ton T. N. B., Tarasenko V. A., Kozyrovskaja N. A. Nitrogen-fixing bacteria colonize rice root xylema. Biopolym. Cell, 1989;5(2):97-99
[3] Belyavskaya N. O., Kozyrovskaya N. O., Kucherenko L. O., Kordyum E. L., Kordyum V. A. Interrelations of the Klebsiella genera with the plant. I. Electron microscopic analysis of endophytic microorganisms interrelationship with rice seedling roots. Biopolym. Cell, 1995;11(1):55-60
[4] Kozyrovskaya N. A., Makitruk V. L., Ruckdashell E. Nitrogen-fixing Klebsiella species produce indole-3-acetic acid Biopolym. Cell, 1990;6(6):93-96
[5] Kozyrovska N., Alexeyev M., Kovtunovych G., Gun'kovska N., Kordyum V. Survival of Klebsiella oxytoca VN13 engineered to bioluminescence on barley roots during plant vegetation. Microb. Releases., 1994; 2:261-265.
[6] Huang J. Ultrastructure of bacterial penetration in plants. Annu. Rev. Phytopathol., 1986; 24(1):141-157.
[7] Starr MP, Chatterjee AK. The genus Erwinia: enterobacteria pathogenic to plants and animals. Annu Rev Microbiol. 1972;26:389-426.
[8] von Riesen VL. Polypectate digestion by Yersinia. J Clin Microbiol. 1975;2(6):552-3.
[9] Von Riesen V.L. Pectinolytic, indole positive strains of Klebsiella pneumoniae. International Journal of Systematic Bacteriology,1976; 26 (2):143-145.
[10] Kuczius T, Kleiner D. Ammonia-excreting mutants of Klebsiella pneumoniae with a pleiotropic defect in nitrogen metabolism. Arch Microbiol. 1996;166(6):388-93.
[11] Miller JH. Experiments in molecular genetics. Cold Spring Harbor Laboratory, 1972, 466 p.
[12] Neuer G., Kronenberg A., Bothe H. Denitrification and nitrogen fixation by Azospirillum. III. Properties of a wheat-Azospirillum association. Archives of Microbiology, 1985;141 (4):364-370.
[13] Wood P.J. The use of dye-polysaccharide interactions in ?-d-glucanase assay. Carbohydrate Research, 1981;94 (2), pp. C19-C23.
[14] Ghose T., Montenecourt B.S., Eveleigh D.E. Measure of cellulase activity (substrates, activities and recommendations). Preprint of JUP AC Commission on Biotechnology 1981.
[15] Cuppels D, Kelman A. Evaluation of selective media for isolation of soft-rot bacteria from soil and plant tissue. Phytopathology., 1973; 64(4):468-475.
[16] Starr MP, Chatterjee AK, Starr PB, Buchanan GE. Enzymatic degradation of polygalacturonic acid by Yersinia and Klebsiella species in relation to clinical laboratory procedures. J Clin Microbiol. 1977;6(4):379-86.
[17] Reinhold-Hurek B., Hurek T. Capacity of Azoarcus. New horizons in nitrogen fixation: Proc. 9th Int. Congr. Nitrogen Fixation, (6-12 December 1992, Cancun). Eds R. Palasios, J. Mora and W. E. Newton.-Mexico; Kluwer; Dordrecht 1992. 671-675.
[18] Reinhold-Hurek B, Hurek T, Claeyssens M, van Montagu M. Cloning, expression in Escherichia coli, and characterization of cellulolytic enzymes of Azoarcus sp., a root-invading diazotroph. J Bacteriol. 1993;175(21):7056-65.
[19] Shishido M, Loeb BM, Chanway CP. External and internal root colonization of logepole pine seedlings by two growthpromoting Bacillus strains originated from different root microsites. Can. J. Microbiol., 1995;41(8): 707-713.
[20] Quadt-Hallmann A, Benhamou N, Kloepper JW. Bacterial endophytes in cotton: Mechanisms of entering the plant. Canadian Journal of Microbiology, 1997; 43 (6):577-582.
[21] Chatterjee AK, Buchanan GE, Behrens MK, Starr MP. Synthesis and excretion of polygalacturonic acid trans-eliminase in Erwinia, Yersinia, and Klebsiella species. Can J Microbiol. 1979;25(1):94-102.
[22] Petak A. M., Kovtunovich G. L., Kozyrovskaya N. A., Turyanitsa A. I., Kordyum V. A. Interrelations of the Klebsiella genera with the plant. 2. Localization of K. oxytoca and K. terriaena into the tobacco and wheat tissues Biopolym. Cell, 1995;11(6):75-80