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. 1994 Jul;60(7):2278–2285. doi: 10.1128/aem.60.7.2278-2285.1994

Derivation of Mutants of Erwinia carotovora subsp. betavasculorum Deficient in Export of Pectolytic Enzymes with Potential for Biological Control of Potato Soft Rot

José M Costa 1, Joyce E Loper 2,*
PMCID: PMC201644  PMID: 16349316

Abstract

Erwinia carotovora subsp. betavasculorum Ecb168 produces an antibiotic(s) that suppresses growth of the related bacterium Erwinia carotovora subsp. carotovora in culture and in wounds of potato tubers. Strain Ecb168 also produces and secretes pectolytic enzymes and causes a vascular necrosis and root rot of sugar beet. Genes (out) involved in secretion of pectolytic enzymes by Ecb168 were localized to two HindIII fragments (8.5 and 10.5 kb) of Ecb168 genomic DNA by hybridization to the cloned out region of E. carotovora subsp. carotovora and by complementation of Out- mutants of E. carotovora subsp. carotovora. Out- mutants of Ecb168, which did not secrete pectate lyase into the culture medium, were obtained when deletions internal to either HindIII fragment were introduced into the genome of Ecb168 through marker exchange mutagenesis. Out- mutants of Ecb168 were complemented to the Out+ phenotype by introduction of the corresponding cloned HindIII fragment. Out- mutants of Ecb168 were less virulent than the Out+ parental strain on potato tubers. Strain Ecb168 and Out- derivatives inhibited the growth of E. carotovora subsp. carotovora in culture, indicating that the uncharacterized antibiotic(s) responsible for antagonism was exported through an out-independent mechanism. Strain Ecb168 and Out- derivatives reduced the establishment of large populations of E. carotovora subsp. carotovora in wounds of potato tubers and suppressed tuber soft rot caused by E. carotovora subsp. carotovora.

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Selected References

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  1. Andro T., Chambost J. P., Kotoujansky A., Cattaneo J., Bertheau Y., Barras F., Van Gijsegem F., Coleno A. Mutants of Erwinia chrysanthemi defective in secretion of pectinase and cellulase. J Bacteriol. 1984 Dec;160(3):1199–1203. doi: 10.1128/jb.160.3.1199-1203.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axelrood P. E., Rella M., Schroth M. N. Role of antibiosis in competition of erwinia strains in potato infection courts. Appl Environ Microbiol. 1988 May;54(5):1222–1229. doi: 10.1128/aem.54.5.1222-1229.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Butler W., Cook L., Vayda M. E. Hypoxic stress inhibits multiple aspects of the potato tuber wound response. Plant Physiol. 1990 May;93(1):264–270. doi: 10.1104/pp.93.1.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Figurski D. H., Helinski D. R. 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 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jones S., Yu B., Bainton N. J., Birdsall M., Bycroft B. W., Chhabra S. R., Cox A. J., Golby P., Reeves P. J., Stephens S. The lux autoinducer regulates the production of exoenzyme virulence determinants in Erwinia carotovora and Pseudomonas aeruginosa. EMBO J. 1993 Jun;12(6):2477–2482. doi: 10.1002/j.1460-2075.1993.tb05902.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Keen N. T., Tamaki S. Structure of two pectate lyase genes from Erwinia chrysanthemi EC16 and their high-level expression in Escherichia coli. J Bacteriol. 1986 Nov;168(2):595–606. doi: 10.1128/jb.168.2.595-606.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lei S. P., Lin H. C., Heffernan L., Wilcox G. Cloning of the pectate lyase genes from Erwinia carotovora and their expression in Escherichia coli. Gene. 1985;35(1-2):63–70. doi: 10.1016/0378-1119(85)90158-1. [DOI] [PubMed] [Google Scholar]
  8. Lindeberg M., Collmer A. Analysis of eight out genes in a cluster required for pectic enzyme secretion by Erwinia chrysanthemi: sequence comparison with secretion genes from other gram-negative bacteria. J Bacteriol. 1992 Nov;174(22):7385–7397. doi: 10.1128/jb.174.22.7385-7397.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Loper J. E., Haack C., Schroth M. N. Population Dynamics of Soil Pseudomonads in the Rhizosphere of Potato (Solanum tuberosum L.). Appl Environ Microbiol. 1985 Feb;49(2):416–422. doi: 10.1128/aem.49.2.416-422.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Murata H., Fons M., Chatterjee A., Collmer A., Chatterjee A. K. Characterization of transposon insertion out- mutants of Erwinia carotovora subsp. carotovora defective in enzyme export and of a DNA segment that complements out mutations in E. carotovora subsp. carotovora, E. carotovora subsp. atroseptica, and Erwinia chrysanthemi. J Bacteriol. 1990 Jun;172(6):2970–2978. doi: 10.1128/jb.172.6.2970-2978.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Reeves P. J., Whitcombe D., Wharam S., Gibson M., Allison G., Bunce N., Barallon R., Douglas P., Mulholland V., Stevens S. Molecular cloning and characterization of 13 out genes from Erwinia carotovora subspecies carotovora: genes encoding members of a general secretion pathway (GSP) widespread in gram-negative bacteria. Mol Microbiol. 1993 May;8(3):443–456. doi: 10.1111/j.1365-2958.1993.tb01589.x. [DOI] [PubMed] [Google Scholar]
  12. Rella M., Axelrood P. E., Weinhold A. R., Schroth M. N. Improvement of DNA transfer frequency and transposon mutagenesis of Erwinia carotovora subsp. betavasculorum. Appl Environ Microbiol. 1989 Apr;55(4):934–939. doi: 10.1128/aem.55.4.934-939.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ried J. L., Collmer A. An nptI-sacB-sacR cartridge for constructing directed, unmarked mutations in gram-negative bacteria by marker exchange-eviction mutagenesis. Gene. 1987;57(2-3):239–246. doi: 10.1016/0378-1119(87)90127-2. [DOI] [PubMed] [Google Scholar]
  14. Roberts D. P., Berman P. M., Allen C., Stromberg V. K., Lacy G. H., Mount M. S. Requirement for two or more Erwinia carotovora subsp. carotovora pectolytic gene products for maceration of potato tuber tissue by Escherichia coli. J Bacteriol. 1986 Jul;167(1):279–284. doi: 10.1128/jb.167.1.279-284.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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 Dec;169(12):5789–5794. doi: 10.1128/jb.169.12.5789-5794.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Thurn K. K., Chatterjee A. K. Single-site chromosomal Tn5 insertions affect the export of pectolytic and cellulolytic enzymes in Erwinia chrysanthemi EC16. Appl Environ Microbiol. 1985 Oct;50(4):894–898. doi: 10.1128/aem.50.4.894-898.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]

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