Trehalose Biosynthesis Promotes <em>Pseudomonas aeruginosa</em> Pathogenicity in Plants

<div><p><i>Pseudomonas aeruginosa</i> strain PA14 is a multi-host pathogen that infects plants, nematodes, insects, and vertebrates. Many PA14 factors are required for virulence in more than one of these hosts. Noting that plants have a fundamentally different cellular architecture from animals, we sought to identify PA14 factors that are specifically required for plant pathogenesis. We show that synthesis by PA14 of the disaccharide trehalose is required for pathogenesis in Arabidopsis, but not in nematodes, insects, or mice. In-frame deletion of two closely-linked predicted trehalose biosynthetic operons, <i>treYZ</i> and <i>treS</i>, decreased growth in Arabidopsis leaves about 50 fold. Exogenously co-inoculated trehalose, ammonium, or nitrate, but not glucose, sulfate, or phosphate suppressed the phenotype of the double Δ<i>treYZ</i>Δ<i>treS</i> mutant. Exogenous trehalose or ammonium nitrate does not suppress the growth defect of the double Δ<i>treYZ</i>Δ<i>treS</i> mutant by suppressing the plant defense response. Trehalose also does not function intracellularly in <i>P. aeruginosa</i> to ameliorate a variety of stresses, but most likely functions extracellularly, because wild-type PA14 rescued the <i>in vivo</i> growth defect of the Δ<i>treYZ</i>Δ<i>treS</i> in trans. Surprisingly, the growth defect of the double Δ<i>treYZ</i>Δ<i>treS</i> double mutant was suppressed by various Arabidopsis cell wall mutants that affect xyloglucan synthesis, including an <i>xxt1xxt2</i> double mutant that completely lacks xyloglucan, even though xyloglucan mutants are not more susceptible to pathogens and respond like wild-type plants to immune elicitors. An explanation of our data is that trehalose functions to promote the acquisition of nitrogen-containing nutrients in a process that involves the xyloglucan component of the plant cell wall, thereby allowing <i>P. aeruginosa</i> to replicate in the intercellular spaces in a leaf. This work shows how <i>P. aeruginosa</i>, a multi-host opportunistic pathogen, has repurposed a highly conserved “house-keeping” anabolic pathway (trehalose biosynthesis) as a potent virulence factor that allows it to replicate in the intercellular environment of a leaf.</p>