Application of Chemical Genomics to Plant–Bacteria Communication: A High-Throughput System to Identify Novel Molecules Modulating the Induction of Bacterial Virulence Genes by Plant Signals
The life cycle of bacterial phytopathogens consists of a benign epiphytic phase, during which the bacteria grow in the soil or on the plant surface, and a virulent endophytic phase involving the penetration of host defenses and the colonization of plant tissues. Innovative strategies are urgently required to integrate copper treatments that control the epiphytic phase with complementary tools that control the virulent endophytic phase, thus reducing the quantity of chemicals applied to economically and ecologically acceptable levels. Such strategies include targeted treatments that weaken bacterial pathogens, particularly those inhibiting early infection steps rather than tackling established infections. This chapter describes a reporter gene-based chemical genomic high-throughput screen for the induction of bacterial virulence by plant molecules. Specifically, we describe a chemical genomic screening method to identify agonist and antagonist molecules for the induction of targeted bacterial virulence genes by plant extracts, focusing on the experimental controls required to avoid false positives and thus ensuring the results are reliable and reproducible.
Key wordsPhytopathogenic bacterial virulence Plant extract Reporter gene Virulence gene promoter activity Chemical library High-throughput screening
This work was supported by a grant from the Italian Veneto region “Progetto di innovazione per la difesa della pianta del kiwi e per la valorizzazione dei suoi frutti” (DGR n. 2587–23/12/2014). Work in AC’s laboratory was funded by the Spanish Ministry for Science and Innovation grant BIO2013–44407-R. We thank Prof. Scortichini (CREA-FRC, Caserta, Italy) for providing the strain of Pseudomonas syringae pv. actinidiae CRAFRU8.43.
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