Synthetic dual hormone-responsive promoters enable engineering of plants with broad-spectrum resistance.

In plant immunity, the mutually antagonistic hormones salicylic acid (SA) and jasmonic acid (JA) are implicated in resistance to biotrophic and necrotrophic pathogens, respectively. Promoters that can respond to both SA and JA signals are urgently needed to engineer plants with enhanced resistance to a broad spectrum of pathogens. However, few natural pathogen-inducible promoters are available for this purpose. To address this problem, we have developed a strategy to synthesize dual SA- and JA-responsive promoters by combining SA- and JA-responsive cis elements based on the interaction between their cognate trans-acting factors. The resulting promoters respond rapidly and strongly to both SA and Methyl Jasmonate (MeJA), as well as different types of phytopathogens. When such a synthetic promoter was used to control expression of an antimicrobial peptide, transgenic plants displayed enhanced resistance to a diverse range of biotrophic, necrotrophic, and hemi-biotrophic pathogens. A dual-inducible promoter responsive to the antagonistic signals auxin and cytokinin was generated in a similar manner, confirming that our strategy can be used for the design of other biotically or abiotically inducible systems.

The role of urease in the acid stress response and fimbriae expression in Klebsiella pneumoniae CG43.

BACKGROUND/PURPOSE: Two urease operons were identified in Klebsiella pneumoniae CG43, ure-1 and ure-2. This study investigates whether a differential regulation of the expression of ure-1 and ure-2 exists and how urease activity influences the acid stress response and expression of type 1 and type 3 fimbriae. METHODS: The ureA1 and ureA2 gene specific deletion mutants were constructed. Promoter activity was assessed using a LacZ reporter system. The sensitivity to acid stress was determined by assessing the survival after pH 2.5 treatment. The influence on type 1 and type 3 fimbriae expression was assessed using western blotting and mannose-sensitive yeast agglutination and biofilm formation assay, respectively. RESULTS: Bacterial growth analysis in mM9-U or modified Stuart broth revealed that ure-1 was the principal urease system, and ure-2 had a negative effect on ure-1 activity. Deletion of the fur or nac gene had no apparent effect on the activity of Pure1, Pure2-1, and Pure2-2. The Pure2-2 activity was enhanced by deletion of the hns gene. ureA1 deletion increased acid stress sensitivity, whereas the deleting effect of ureA2 was notable without hns. Deletion of ureA1 or ureA2 significantly induced the expression of type 1 fimbriae but decreased MrkA production and biofilm formation. CONCLUSION: ure-1 is the primary expression system in K. pneumoniae CG43, while ure-2 is active in the absence of hns. Impairment of urease activity increases the sensitivity to acid stress, and the accumulation of urea induces the expression of type 1 fimbriae but represses type 3 fimbriae expression.