title: Characterization of a Ralstonia solanacearum operon required for polygalacturonate degradation and uptake of galacturonic acid.
abstract: The bacterial wilt pathogen Ralstonia solanacearum produces three extracellular polygalacturonases (PGs): PehA, PehB, and PehC. All three PGs hydrolyze pectin's polygalacturonic acid backbone, but each releases different reaction products. PehA and PehB contribute significantly to pathogen virulence, probably by facilitating root invasion and colonization. To determine the collective contribution of PGs to virulence and saprophytic survival, we cloned, characterized, and mutated the R. solanacearum pehC gene, which encodes a distinctive monogalacturonate-releasing exo-PG. The virulence of a pehC mutant on tomato was indistinguishable from that of its wild-type parent; thus, this exo-PG alone does not contribute significantly to wilt pathogenesis. Unexpectedly, a completely PG-deficient triple pehA/B/C mutant was slightly more virulent than a pehA/B mutant. PehC may degrade galacturonide elicitors of host defense, thereby protecting the pathogen from plant antimicrobial responses. A galacturonate transporter gene, exuT, is immediately downstream of pehC and the two genes are co-transcribed. It has been hypothesized that galacturonic acid released by PGs from plant cell walls nourishes bacteria during pathogenesis. To separate the pectolytic and nutrient-generating roles of the PGs, we made an exuT mutant, which still produces all three isozymes of PG but cannot uptake PG degradation products. This exuT mutant had wild-type virulence on tomato, demonstrating that metabolism of galacturonic acid does not contribute significantly to bacterial success inside the plant.
authors.affiliation: Department of Plant Pathology, University of Wisconsin-Madison, 1630 Linden Drive, Madison 53706, USA.
authors.forename: Enid T
authors.initials: ET
authors.lastname: González
authors.forename: Caitilyn
authors.initials: C
authors.lastname: Allen
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000378:metabolism; D016946:Gram-Negative Aerobic Rods and Cocci / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D006603:Hexuronic Acids / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon / Q000235:genetics*; D010368:Pectins / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C111694:ExuT protein, bacteria; D006603:Hexuronic Acids; D008565:Membrane Proteins; C007819:galacturonic acid; D010368:Pectins; D006026:Glycoside Hydrolases; C020136:exopolygalacturonase; C003181:polygalacturonic acid