Pseudomonas corrugata crpCDE is part of the cyclic lipopeptide corpeptin biosynthetic gene cluster and is involved in bacterial virulence in tomato and in hypersensitive response in Nicotiana benthamiana

Molecular Plant Pathology
2015.0

Abstract

<jats:title>Summary</jats:title><jats:p><jats:italic><jats:styled-content style="fixed-case">P</jats:styled-content>seudomonas corrugata</jats:italic> <jats:styled-content style="fixed-case">CFBP</jats:styled-content> 5454 produces two kinds of cyclic lipopeptides (<jats:styled-content style="fixed-case">CLP</jats:styled-content>s), cormycin <jats:styled-content style="fixed-case">A</jats:styled-content> and corpeptins, both of which possess surfactant, antimicrobial and phytotoxic activities. In this study, we identified genes coding for a putative non‐ribosomal peptide synthetase and an <jats:styled-content style="fixed-case">ABC</jats:styled-content>‐type transport system involved in corpeptin production. These genes belong to the same transcriptional unit, designated <jats:italic><jats:styled-content style="fixed-case">crpCDE</jats:styled-content></jats:italic>. The genetic organization of this locus is highly similar to other <jats:italic><jats:styled-content style="fixed-case">P</jats:styled-content>seudomonas</jats:italic> <jats:styled-content style="fixed-case">CLP</jats:styled-content> biosynthetic clusters. Matrix‐assisted laser desorption ionization‐time of flight‐mass spectrometry (<jats:styled-content style="fixed-case">MALDI‐TOF</jats:styled-content>‐<jats:styled-content style="fixed-case">MS</jats:styled-content>) analysis revealed that transporter and synthetase genomic knock‐out mutants were unable to produce corpeptins, but continued to produce cormycin <jats:styled-content style="fixed-case">A</jats:styled-content>. This suggests that <jats:styled-content style="fixed-case">CrpCDE</jats:styled-content> is the only system involved in corpeptin production in <jats:italic><jats:styled-content style="fixed-case">P</jats:styled-content>. corrugata</jats:italic> <jats:styled-content style="fixed-case">CFBP</jats:styled-content> 5454. In addition, phylogenetic analysis revealed that the <jats:styled-content style="fixed-case">CrpE ABC</jats:styled-content> transporter clustered with the transporters of <jats:styled-content style="fixed-case">CLPs</jats:styled-content> with a long peptide chain. Strains depleted in corpeptin production were significantly less virulent than the wild‐type strain when inoculated in tomato plants and induced only chlorosis when infiltrated into <jats:italic><jats:styled-content style="fixed-case">N</jats:styled-content>icotiana benthamiana</jats:italic> leaves. Thus, corpeptins are important effectors of <jats:italic><jats:styled-content style="fixed-case">P</jats:styled-content>. corrugata</jats:italic> interaction with plants. Expression analysis revealed that <jats:italic><jats:styled-content style="fixed-case">crpC</jats:styled-content></jats:italic> transcription occurs at high cell density. Two <jats:styled-content style="fixed-case">LuxR</jats:styled-content> transcriptional regulators, <jats:styled-content style="fixed-case">PcoR</jats:styled-content> and <jats:styled-content style="fixed-case">RfiA</jats:styled-content>, have a pivotal role in <jats:italic><jats:styled-content style="fixed-case">crpC</jats:styled-content></jats:italic> expression and thus in corpeptin production.

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