Abstract
Formation of nodules on roots or in stems (in some cases) of leguminous plants is the unique ability of gram-negative bacteria, Rhizobia, which converts atmospheric nitrogen into usable forms by the host plant. Lipopolysaccharide (LPS) is the outer membrane component of the gram-negative bacteria, known to be an essential factor in host recognition, specificity, and initial infection processes. In the present study, we extracted lipopolysaccharides from different rhizobial isolates by a modified phenol-water method and partially characterized by polyacrylamide gel electrophoresis with silver staining. The results showed two separate banding regions, LPS-I and LPS-II. The high molecular weight and electrophoretic mobility of LPS-I region resembles that of lysozyme, used as a standard marker. The LPS-II region has a low molecular weight and electrophoretic mobility greater than that of lysozyme. The LPS-II region was due to incomplete LPS, which either lacks the entire O-antigen repeating unit or contains only one or two repeating units. The banding patterns of LPS vary among the different rhizobial isolates. Results revealed that the type of LPS structure and banding regions greatly facilitate the further characterization of the LPS modifications required for symbiosis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bauer WD. 1981. Infection of legumes by rhizobia. Annu. Rev. Plant Physiol. 32: 407–449
Brewin NJ. 1998. Tissue and cell invasion by Rhizobium: the structure and development of infection threads and symbio somes,. In HP Spaink, A Kondorosi, PJJ Hooykaas, eds, The Rhizobiaceae: Molecular Biology of Model Plant-Associated Bacteria, Kluwer Academic Publishers, Dordecht, pp 417–429
Brewin NJ, Wood EA, Larkins AP, Galfre G, Butcher GW. 1986. Analysis of lipopolysaccharide from root nodule bac teroids of Rhizobium leguminosarum using monoclonal anti bodies. J. Gen. Microbiol. 132: 1959–1968
Carlson RW. 1982. Surface chemistry, In WJ Broughton, ed, Nitrogen Fixation, Vol 2, Rhizobium, Clarendon Press, Oxford, pp 199–234
Carlson RW. 1984. The heterogeneity of Rhizobium lipopolysaccharides. J. Bacteriol. 158: 1012–1017
Carlson RW, Lee R. 1983. A comparison of the surface polysac charides from Rhizobium leguminosarum 128C53smrr rif with the surface polysaccharides from its exo-1 mutant. Plant Physiol. 71: 223–228
Carlson RW, Sanders RE, Napoli C, Albersheim P. 1978. Hostsymbiont interactions III. Purification and characterization of Rhizobium lipopolysaccharides. Plant Physiol. 62: 912–917
Cava JR, Elias PM, Turowski DA, Noel KD. 1989. Rhizobium leguminosarum CFN42 genetic regions encoding lipopolysaccharide structures essential for complete nodule development on bean plants. J. Bacteriol. 171: 8–15
Dazzo FB, Truchet GL, Hollingsworth RI, Hrabak EM, Pankratz, HS, Philip-Hollingsworth S, Salzwedel JL, Chapaman K, Appenzeller L, Squartini A, Gerhold D, Orgambide G. 1991. Rhizobium lipopolysaccharide modu lates infection thread development in white clover root hairs. J. Bacteriol. 173: 5371–5384
de Maagd RA, Rao AS, Mulders IHM, Goosen-de Roo L, van Loosdrech MCM, Wiffelman CA, Lugtenberg BJJ. 1989. Isolation and characterization of mutants of Rhizobium legu minosarum bv. viciae 248 with altered lipopolysaccharides: possible role of surface charge or hydrophobicity in bacterial release from the infection thread. J. Bacteriol. 171: 1143–1150
D’Haeze W, Leoff C, Freshour G, Noel KD, Carlson RW. 2007. Rhizobium etli CE3 bacteroid lipopolysaccharides are struc turally similar but not identical to those produced by cultured CE3 bacteria. J. Biol. Chem. 282: 17101–17113
Fomsgaard A, Freudenber MA, Galanos C. 1990. Modification of the silver staining technique to detect lipopolysaccharide in polyacrylamide gels. J. Clin. Microbiol. 28: 2627–2631
Forsberg LS, Carlson RW. 2008. Structural characterization of the primary O-antigenic polysaccharide of the Rhizobium leguminosarum 3841 lipopolysaccharide: isolation and identi fication of a new 3-acetimidoylamino-3-deoxyhexuronic acid glycosyl component. J. Biol. Chem. 283: 16037–16050
Jabbouri S, Hannin M, Fellay R, Quesada-Vincens D, Ruehs BL, Carlson RW, Broughton WJ. 1996. Rhizobium species NGR234 host-specificity of nodulation locus III contains nod and fix genes. In G Stacey, B Mullin, PM Gresshoff, eds Biology of Plant-Microbe Interactions, Intl. Society Plant-Microbe Interact. St. Paul, pp. 319–324
Jadhav SD. 2005. Assessment of biodiversity of native Rhizobia using morphological, biochemical and molecular techniques. M. Sc. thesis submitted in Indira Gandhi Agricultural University, Raipur
Kalia A, Gupta RP. 2002. Nodule induction in non-legume by Rhizobia. Indian J. Microbiol. 42: 183–193
Kannenberg EL, Carlson RW. 2001. Lipid A and O-chain modi fications cause Rhizobium lipopolysaccharides to become hydrophobic during bacteroid development. Mol. Microbiol. 39: 379–391
Kesawat M.S, Manorama, Bhaganagare GR. 2007. Role of rhi zobial lipopolysaccharide in host specificity. Asian J. Microbiol. Biotech. Environ. Sci. 9: 959–964
Noel KD, VandenBosch KA, Kulpaca B. 1986. Mutations in Rhizobium phaseoli that lead to arrested development of infection threads. J. Bacteriol. 168: 1392–1401
Puvanesarajah V, Schell FM, Gerhold D, Stacey G. 1987. Cell surface polysaccharides from Bradyrhizobium japonicum and a nonnodulating mutant. J. Bacteriol. 169: 137–141
Reuhs BL, Geller DP, Kim JS, Fox JE, Kolli VS, Pueppke SG. 1998. Sinorhizobium fredii and Sinorhizobium meliloti pro duce structurally conserved lipopolysaccharides and strainspecific K antigens. Appl. Environ. Microbiol. 64: 4930–4938
Schmidt EL. 1979. Initiation of plant root-microbe interactions. Annu. Rev. Microbiol. 33: 355–376
Song SC, Lin LP. 1999. The transition of Rhizobium fredii lipopolysaccharides induced by soybean root exudation. Bot. Bull. Acad. Sin. 40: 73–78
Summers ML, Botero LM, Busse SC, Mcdermott TR. 2000. The Sinorhizobium meliloti Lon protease is involved in regulating exopolysaccharide synthesis and is required for nodulation of alfalfa. J. Bacteriol. 182: 2551–2558
Tsai CM, Frasch CE. 1982. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Ann. Biochem. 119: 115–119
Zevenhuizen LPTM, Scholten-Koerselman I, Posthumus MA. 1980. Lipopolysaccharides of Rhizobium. Arch. Microbiol. 125: 1–8
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kesawat, M.S., Das, B.K., Bhaganagare, G.R. et al. Isolation and characterization of lipopolysaccharides from different rhizobial isolates. J. Crop Sci. Biotechnol. 12, 109–113 (2009). https://doi.org/10.1007/s12892-009-0101-0
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12892-009-0101-0