Genetic and Biochemical Characterization of a Pseudomonas Solanacearum Gene Cluster Required for Extracellular Polysaccharide Production and for Virulence

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1991 Mar 1

PMID 1999385

Citations 14

Authors

D Cook

L Sequeira

Affiliations

Soon will be listed here.

Abstract

Infection of host plants by Pseudomonas solanacerum results in wilting, which is thought to be due largely to the occlusion of xylem vessels by the P. solanacearum extracellular polysaccharide (EPS) that primarily consists of N-acetylgalactosamine (GalNAc). By means of Tn3 mutagenesis, we identified a 6.5-kb gene cluster that contains five complementation units required for EPS production and virulence in this bacterium. There was positive correlation between the amount of EPS produced in culture and (i) in planta growth and (ii) virulence. Based on analysis of beta-glucuronidase-gene fusions, these genes are expressed both in broth cultures and in planta and may be constitutive. Both wild-type and mutant strains contained similar amounts of UDP-GalNAc, the predicted primary substrate for EPS synthesis. Thus, the EPS mutants we obtained should be useful in the analysis of steps in the assembly of the polysaccharide and how this process is related to virulence.

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References

Kelman A, Hruschka J . The role of motility and aerotaxis in the selective increase of avirulent bacteria in still broth cultures of Pseudomonas solanacearum. J Gen Microbiol. 1973; 76(1):177-88. DOI: 10.1099/00221287-76-1-177. View

Harding N, Cleary J, Cabanas D, Rosen I, Kang K . Genetic and physical analyses of a cluster of genes essential for xanthan gum biosynthesis in Xanthomonas campestris. J Bacteriol. 1987; 169(6):2854-61. PMC: 212199. DOI: 10.1128/jb.169.6.2854-2861.1987. View

Drigues P, Trigalet A, Dupin P, Samain D, Asselineau J . Comparative studies of lipopolysaccharide and exopolysaccharide from a virulent strain of Pseudomonas solanacearum and from three avirulent mutants. J Bacteriol. 1985; 162(2):504-9. PMC: 218876. DOI: 10.1128/jb.162.2.504-509.1985. View

Dolph P, Majerczak D, Coplin D . Characterization of a gene cluster for exopolysaccharide biosynthesis and virulence in Erwinia stewartii. J Bacteriol. 1988; 170(2):865-71. PMC: 210734. DOI: 10.1128/jb.170.2.865-871.1988. View

Baker C, Neilson M, Sequeira L, Keegstra K . Chemical Characterization of the Lipopolysaccharide of Pseudomonas solanacearum. Appl Environ Microbiol. 1984; 47(5):1096-100. PMC: 240067. DOI: 10.1128/aem.47.5.1096-1100.1984. View

Xu P, Iwata M, Leong S, Sequeira L . Highly virulent strains of Pseudomonas solanacearum that are defective in extracellular-polysaccharide production. J Bacteriol. 1990; 172(7):3946-51. PMC: 213378. DOI: 10.1128/jb.172.7.3946-3951.1990. View

Van Alfen N, McMillan B, Turner V, Hess W . Role of pit membranes in macromolecule-induced wilt of plants. Plant Physiol. 1983; 73(4):1020-3. PMC: 1066599. DOI: 10.1104/pp.73.4.1020. View

Tolmasky M, Staneloni R, LELOIR L . Lipid-bound saccharides in Rhizobium meliloti. J Biol Chem. 1982; 257(12):6751-7. View

Stachel S, An G, Flores C, Nester E . A Tn3 lacZ transposon for the random generation of beta-galactosidase gene fusions: application to the analysis of gene expression in Agrobacterium. EMBO J. 1985; 4(4):891-8. PMC: 554276. DOI: 10.1002/j.1460-2075.1985.tb03715.x. View

10.

Easson Jr D, Sinskey A, Peoples O . Isolation of Zoogloea ramigera I-16-M exopolysaccharide biosynthetic genes and evidence for instability within this region. J Bacteriol. 1987; 169(10):4518-24. PMC: 213816. DOI: 10.1128/jb.169.10.4518-4524.1987. View

11.

Hotte B, Puhler A, Simon R . Cloning and analysis of a 35.3-kilobase DNA region involved in exopolysaccharide production by Xanthomonas campestris pv. campestris. J Bacteriol. 1990; 172(5):2804-7. PMC: 208934. DOI: 10.1128/jb.172.5.2804-2807.1990. View

12.

Figurski D, Helinski D . Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979; 76(4):1648-52. PMC: 383447. DOI: 10.1073/pnas.76.4.1648. View

13.

Roberts D, Denny T, Schell M . Cloning of the egl gene of Pseudomonas solanacearum and analysis of its role in phytopathogenicity. J Bacteriol. 1988; 170(4):1445-51. PMC: 210987. DOI: 10.1128/jb.170.4.1445-1451.1988. View

14.

Bochner B, Ames B . Complete analysis of cellular nucleotides by two-dimensional thin layer chromatography. J Biol Chem. 1982; 257(16):9759-69. View

15.

Whiteheart S, Passaniti A, Reichner J, Holt G, Haltiwanger R, Hart G . Glycosyltransferase probes. Methods Enzymol. 1989; 179:82-95. DOI: 10.1016/0076-6879(89)79116-3. View

16.

Xu P, Leong S, Sequeira L . Molecular cloning of genes that specify virulence in Pseudomonas solanacearum. J Bacteriol. 1988; 170(2):617-22. PMC: 210699. DOI: 10.1128/jb.170.2.617-622.1988. View

17.

Amasino R . Acceleration of nucleic acid hybridization rate by polyethylene glycol. Anal Biochem. 1986; 152(2):304-7. DOI: 10.1016/0003-2697(86)90413-6. View

18.

Staskawicz B, Dahlbeck D, Keen N, Napoli C . Molecular characterization of cloned avirulence genes from race 0 and race 1 of Pseudomonas syringae pv. glycinea. J Bacteriol. 1987; 169(12):5789-94. PMC: 214142. DOI: 10.1128/jb.169.12.5789-5794.1987. View

19.

Hendrick C, Sequeira L . Lipopolysaccharide-Defective Mutants of the Wilt Pathogen Pseudomonas solanacearum. Appl Environ Microbiol. 1984; 48(1):94-101. PMC: 240322. DOI: 10.1128/aem.48.1.94-101.1984. View