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The Contribution of Syringopeptin and Syringomycin to Virulence of Pseudomonas syringae pv. syringae strain B301D on the Basis of sypA and syrB1 Biosynthesis Mutant Analysis

March 2001 , Volume 14 , Number  3
Pages  336 - 348

Brenda K. Scholz-Schroeder , 1 Michael L. Hutchison , 1 Ingeborg Grgurina , 2 and Dennis C. Gross 1

1Department of Plant Pathology, Washington State University, Pullman 99164-6430, U.S.A.; 2Dipartmento di Scienze Biochimiche “A. Rossi Fanelli,” Università “La Sapienza” di Rome, Italy

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Accepted 7 November 2000.

Sequencing of an approximately 3.9-kb fragment downstream of the syrD gene of Pseudomonas syringae pv. syringae strain B301D revealed that this region, designated sypA, codes for a peptide synthetase, a multifunctional enzyme involved in the thiotemplate mechanism of peptide biosynthesis. The translated protein sequence encompasses a complete amino acid activation module containing the conserved domains characteristic of peptide synthetases. Analysis of the substrate specificity region of this module indicates that it incorporates 2,3-dehydroaminobutyric acid into the syringopeptin peptide structure. Bioassay and high performance liquid chromatography data confirmed that disruption of the sypA gene in strain B301D resulted in the loss of syringopeptin production. The contribution of syringopeptin and syringomycin to the virulence of P. syringae pv. syringae strain B301D was examined in immature sweet cherry with sypA and syrB1 synthetase mutants defective in the production of the two toxins, respectively. Syringopeptin (sypA) and syringomycin (syrB1) mutants were reduced in virulence 59 and 26%, respectively, compared with the parental strain in cherry, whereas the syringopeptin-syringomycin double mutant was reduced 76% in virulence. These data demonstrate that syringopeptin and syringomycin are major virulence determinants of P. syringae pv. syringae.

Additional keywords: lipopeptide toxin, necrosis-inducing toxin, nonribosomal peptide synthetase, phytotoxin.

© 2001 The American Phytopathological Society