Abstract
Effect of the carbon source in the culture medium and of the growth phase on the composition and structure of the capsular polysaccharides (CPSs) and lipopolysaccharides (LPSs) of the bacterium Azospirillum brasilense Sp245 was studied. Growth with fructose resulted in an increased carbohydrate content in the CPSs, while long-term cultivation resulted in an increased content of phosphorus in both CPSs and LPSs. The LPSs produced on the medium with fructose (regardless of the cultivation duration) and the LPSs of the bacteria grown with sodium malate until the stationary phase were characterized by higher levels of unsaturated fatty acids than the LPSs of the bacteria grown with sodium malate to the late exponential phase. The structures of the polysaccharides from the isolated glycopolymers were established using monosaccharide analysis, including determination of the absolute configurations and 1D and 2D NMR spectroscopy. This study is the first to report that the CPS of A. brasilense Sp245 grown with sodium malate to the end of the exponential phase is structurally identical to the O-polysaccharide from the LPS of this bacterium and that the LPS and CPS of A. brasilense Sp245 grown with fructose contain an additional homoglucan of the following structure: [→3)-α-D-Glcp-(1→] n .
Similar content being viewed by others
References
Altona, C. and Haasnoot, C.A.G., Prediction of anti and gauche vicinal proton-proton coupling constants in carbohydrates: a simple additivity rule for pyranose rings, Org. Magn. Reson., 1980, vol. 13, pp. 417–429.
Bock, K. and Pedersen, C., Carbon-13 nuclear magnetic resonance spectroscopy of monosaccharides, Adv. Carbohydr. Chem. Biochem., 1983, vol. 41, pp. 27–66.
Boyko, A.S., Konnova, S.A., Fedonenko, Yu.P., Zdorovenko, E.L., Smol’kina, O.N., Kachala, V.V., and Ignatov, V.V., Structural and functional peculiarities of the lipopolysaccharide of Azospirillum brasilense SR55, isolated from the roots of Triticum durum, Microbiol. Res., 2011, vol. 166, pp. 585–593.
Burdman, S., Jurkevitch, E., Schwartsburd, B., Hampel, M., and Okon, Y., Aggregation in Azospirillum brasilense: effects of chemical and physical factors and involvement of extracellular components, Microbiology (UK), 1998, vol. 144, pp. 1989–1999.
Das, A. and Mishra, A.K., Utilization of fructose by Azospirillum brasilense, Can. J. Microbiol., 1983, vol. 29, pp. 1213–1217.
Del Gallo, M. and Haegi, A., Characterization and quantification of exocellular polysaccharides in Azospirillum brasilense and Azospirillum lipoferum, Symbiosis, 1990, vol. 9, pp. 155–161.
Egorenkova, I.V., Konnova, S.A., Fedonenko, Yu.P., Dykman, L.A., and Ignatov, V.V., Role of polysaccharide components of Azospirillum brasilense capsules in bacterial adsorption on wheat seedling roots, Microbiology (Moscow), 2001, vol. 70, no. 1, pp. 36–40.
Egorenkova, I.V., Konnova, S.A., Skvortsov, I.M., and Ignatov, V.V., Investigation of the initial stages of interaction of the bacterium Azospirillum brasilense with wheat seedling roots: adsorption and root hair deformation, Microbiology (Moscow), 2000, vol. 69, no. 1, pp. 103–108.
Fedonenko, Yu.P., Sigida, E.N., Konnova, S.A., and Ignatov, V.V., Structure and serology of O-antigens of nitrogenfixing rhizobacteria of the genus Azospirillum, Russ. Chem. Bull., 2015, vol. 64, no. 5, pp. 1024–1031.
Fedonenko, Yu.P., Zatonsky, G.V., Konnova, S.A., Zdorovenko, E.L., and Ignatov, V.V., Structure of the O-specific polysaccharide of the lipopolysaccharide of Azospirillum brasilense Sp245, Carbohydr. Res., 2002, vol. 337, pp. 869–872.
Fibach-Paldi, S., Burdman, S., and Okon, Y., Key physiological properties contributing to rhizosphere adaptation and plant growth promotion abilities of Azospirillum brasilense, FEMS Microbiol. Lett., 2012, vol. 326, pp. 99–108.
Hitchcock, P.J. and Brown, T.M., Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels, J. Bacteriol., 1983, vol. 154, pp. 269–277.
Kannenberg, E.L. and Carlson, R.W., Lipid A and O-chain modifications cause Rhizobium lipopolysaccharides to become hydrophobic during bacteroid development, Mol. Microbiol., 2001, vol. 39, pp. 379–391.
Konnova, O.N., Burygin, G.L., Fedonenko, Yu.P., Matora, L.Yu., Pankin, K.E., Konnova, S.A., and Ignatov, V.V., Chemical composition and immunochemical characteristics of the lipopolysaccharide of nitrogenfixing rhizobacterium Azospirillum brasilense CD, Microbiology (Moscow), 2006, vol. 75, no. 3, pp. 323–328.
Konnova, S.A. and Ignatov, V.V., The chemical composition and functions of Azospirillum extracellular polysaccharidic complexes, in Biology of Plant-Microbe Interaction. Proc. of the 11th Int. Congr. on Molecular Plant-Microbe Interaction, Tikhonovich, I., Lugtenberg, B., and Provorov, N., Eds., S.-Pb., 2003, vol. 4, pp. 327–331.
Konnova, S.A., Brykova, O.S., Sachkova, O.A., Egorenkova, I.V., and Ignatov, V.V., Protective role of the polysaccharide- containing capsular components of Azospirillum brasilense, Microbiology (Moscow), 2001, vol. 70, no. 4, pp. 436–440.
Konnova, S.A., Fedonenko, Yu.P., Makarov, O.E., and Ignatov, V.V., Effect of cultivation conditions on the composition of extracellular polysaccharide-containing substances in the bacterium Azospirillum brasilense, Biol. Bull., 2003, vol. 30, no. 4, pp. 354–360.
Konnova, S.A., Makarov, O.E., Skvortsov, I.M., and Ignatov, V.V., Isolation,fractionation and some properties of polysaccharides produced in a bound form by Azospirillum brasilense and their possible involvement in Azospirillumwheat root interactions, FEMS Microbiol. Lett., 1994, vol. 118, pp. 93–99.
Kravchenko, L.V., Shapozhnikov, A.I., Makarova, N.M., Azarova, T.S., L’vova, K.A., Kostyuk, I.I., Lyapunova, O.A., and Tikhonovich, I.A., Exometabolites of bread wheat and tomato affecting the plant–microbe interactions in the rhizosphere, Russ. J. Plant Physiol., 2011, vol. 58, no. 5, pp. 936–940.
Kul’shin, V.A., Yakovlev, A.P., Avaeva, S.N., and Dmitriev, B.A., Simplified method for lipopolysaccharide isolation from gram-negative bacteria, Mol. Genet. Mikrobiol. Virusol., 1987, no. 5, pp. 44–46.
Leontein, K., Lindberg, B., and Lönngren J., Assignment of absolute configuration of sugars by g.l.c. of their acetylated glycosides formed from chiral alcohols, Carbohydr. Res., 1978, vol. 62, pp. 359–362.
Martinez-Drets, G., Del Gallo, M., Burpee, C., and Burris, R.H., Catabolism of carbohydrates and organic acids and expression of nitrogenase by azospirilla, J. Bacteriol., 1984, vol. 159, pp. 80–85.
Matora, L.Yu. and Shchegolev, S.Yu., Antigenic identity of the capsule lipopolysaccharides, exopolysaccharides, and O-specific polysaccharides in Azospirillum brasilense, Microbiology (Moscow), 2002, vol. 71, no. 2, pp. 178–181.
Mayer, H., Tharanathan, R.N., and Weckesser, J., Analysis of lipopolysaccharides of Gram-negative bacteria, Methods Microbiol., 1985, vol. 18, pp. 157–207.
Michiels, K.W., Croes, C.L., and Vanderleyden, J., Two different modes of attachment of Azospirillum brasilense Sp7 to wheat roots, J. Gen. Microbiol., 1991, vol. 137, pp. 2241–2246.
Myers, M.L. and Hubbell, D.H., Plant cell wall carbohydrates as substrates for Azospirillum brasilense, Appl. Environ. Microbiol., 1987, vol. 53, pp. 2745–2748.
Russo, A., Vettori, L., Felici, C., Fiaschi, G., Morini, S., and Toffanin A., Enhanced micropropagation response and biocontrol effect of Azospirillum brasilense Sp245 on Prunus cerasifera L. clone Mr.S 2/5 plants, J. Biotechnol., 2008, vol. 134, pp. 312–319.
Sadasivan, L. and Neyra, C.A., Flocculation in Azospirillum brasilense and Azospirillum lipoferum: exopolysaccharides and cyst formation, J. Bacteriol., 1985, vol. 163, pp. 716–723.
Sawardeker, J.S., Sloneker, J.H., and Jeanes, A., Quantitative determination of monosaccharides as their alditol acetates by gas liquid chromatography, Anal. Chem., 1965, vol. 37, pp. 1602–1604.
Smol’kina, O.N., Kachala, V.V., Fedonenko, Yu.P., Burygin, G.L., Zdorovenko, E.L., Matora, L.Yu., Konnova, S.A., and Ignatov, V.V., Capsular polysaccharide of the bacterium Azospirillum lipoferum Sp59b: structure and antigenic specificity, Biochemistry (Moscow), 2010, vol. 75, no. 5, pp. 606–613.
Tsai, C.M. and Frasch, C.E., A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels, Anal. Biochem., 1982, vol. 119, pp. 115–119.
Westby, C.A., Cutshall, D.S., and Vigil, G.V., Metabolism of various carbon sources by Azospirillum brasilense, J. Bacteriol., 1983, vol. 156, pp. 1369–1372.
Yevstigneeva, S.S., Khalepa, Ya.V., Sigida, E.N., Fedonenko, Yu.P., Konnova, S.A., and Ignatov, V.V., The influence of different growth conditions on the structure of the bacterial surface glycopolymers of Azospirillum brasilense Sp7, Izv. Saratov. Univ., Ser. Khim. Biol. Ekol., 2014, vol. 14, no.2, pp. 55–61.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © S.S. Yevstigneyeva, E.N. Sigida, Yu.P. Fedonenko, S.A. Konnova, V.V. Ignatov, 2016, published in Mikrobiologiya, 2016, Vol. 85, No. 6, pp. 643–651.
Rights and permissions
About this article
Cite this article
Yevstigneyeva, S.S., Sigida, E.N., Fedonenko, Y.P. et al. Structural properties of capsular and O-specific polysaccharides of Azospirillum brasilense Sp245 under varying cultivation conditions. Microbiology 85, 664–671 (2016). https://doi.org/10.1134/S0026261716060096
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0026261716060096