title: Location and activity of members of a family of virPphA homologues in pathovars of Pseudomonas syringae and P. savastanoi.
abstract: Summary virPphA is a major determinant of the pathogenicity of Pseudomonas savastanoi pv. phaseolicola to Phaseolus bean. A family of homologues of virPphA was detected in pathovars of P. savastanoi and P. syringae. We examined the structure and activity of alleles designated virPphA, virPphA(Pgy), and virPphA(Psv) from P. savastanoi pathovars phaseolicola, glycinea, and savastanoi, respectively, and avrPtoB from P. syringae pv. tomato. Sequencing showed that the virPphA(Pgy) homologue had a 48-bp central deletion in the open reading frame (ORF) compared with virPphA and virPphA(Psv), but otherwise all three P. savastanoi alleles had > 98% identity at the DNA level. By contrast, AvrPtoB from P. syringae pv. tomato strain DC3000 was predicted to have only 51% amino acid similarity with VirPphA. All ORFs have an upstream hrp-box promoter indicating potential regulation by HrpL. Each cloned homologue was introduced into the P. savastanoi pv. phaseolicola strain RW60, which lacks a native plasmid carrying virPphA as part of a pathogenicity island (PAI), and which is not pathogenic on bean. The homologues all restored virulence, as measured by the development of water-soaked lesions in bean pods, and increased bacterial populations in leaves compared with RW60 alone. RW60 harbouring virPphA or virPphA(Psv) elicited a strong hypersensitive reaction (HR) in soybean cv. Osumi; the presence of avrPtoB caused a weak HR, but virPphA(Pgy) did not affect the null reaction observed in soybean with RW60 alone. A second effector gene, avrPphD, was detected on the genomic clones carrying virPphA(Pgy) and virPphA(Psv). avrPphD was also present in both P. savastanoi pv. phaseolicola and P. syringae pv. tomato, but elsewhere in their genomes. Comparison of the genomic locations of virPphA and other effectors found in the P. savastanoi pv. phaseolicola PAI revealed the greatest divergence of the sequences surrounding virPphA to be in P. syringae pv. tomato.
authors.affiliation: Centre for Research in Plant Science, Faculty of Applied Sciences, University of the West of England, Coldharbour Lane, Bristol BS16 1QY, UK.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Jackson
authors.forename: John W
authors.initials: JW
authors.lastname: Mansfield
authors.forename: Hassan
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authors.lastname: Ammouneh
authors.forename: Lindsay C
authors.initials: LC
authors.lastname: Dutton
authors.forename: Becky
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authors.lastname: Wharton
authors.forename: Amaya
authors.initials: A
authors.lastname: Ortiz-Barredo
authors.forename: Dawn L
authors.initials: DL
authors.lastname: Arnold
authors.forename: George
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authors.lastname: Tsiamis
authors.forename: Ane
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authors.lastname: Sesma
authors.forename: Diane
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authors.lastname: Butcher
authors.forename: Jens
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authors.lastname: Boch
authors.forename: Young Jin
authors.initials: YJ
authors.lastname: Kim
authors.forename: Gregory B
authors.initials: GB
authors.lastname: Martin
authors.forename: Stefania
authors.initials: S
authors.lastname: Tegli
authors.forename: Jesús
authors.initials: J
authors.lastname: Murillo
authors.forename: Alan
authors.initials: A
authors.lastname: Vivian