Methods to study PAMP-triggered immunity using tomato and nicotiana benthamiana

Hanh P. Nguyen, Suma Chakravarthy, André C. Velásquez, Heather L. McLane, Lirong Zeng, Hitoshi Nakayashiki, Duck Hwan Park, Alan Collmer, Gregory B. Martin

Research output: Contribution to journalArticle

97 Scopus citations

Abstract

Understanding the molecular basis of plant responses to pathogen-associated molecular patterns (PAMPs) is an active area of research in the field of plant-microbe interactions. A growing number of plant genes involved in various steps of PAMP-triggered immunity (PTI) pathways and microbial factors involved in the elicitation or suppression of PTI have been identified. These studies have largely relied on Arabidopsis thaliana and, therefore, most of the PTI assays have been developed and optimized for that model plant system. Although PTI is a conserved feature among plants, the response spectra vary across different species. Thus, there is a need for robust PTI assays in other pathos v stems, such as those involving Solanaceae plantpathogen interactions, which include many economically important plants and their diseases. We have optimized molecular, cellular, and whole-plant methods to measure PTI responses in two widely studied solanaceous species, tomato (Solanum lycopersicum) and Nicotiana benthamiana. Here, we provide detailed protocols for measuring various PTI-associated phenotypes, including bacterial populations after pretreatment of leaves with PAMPs, induction of reporter genes, callose deposition, activation of mitogenactivated protein kinases, and a luciferase-based reporter system. These methods will facilitate limited genetic screens and detailed characterization of potential PTI-related genes in model and economically important Solanaceae spp.-pathogen interactions.

Original languageEnglish (US)
Pages (from-to)991-999
Number of pages9
JournalMolecular Plant-Microbe Interactions
Volume23
Issue number8
DOIs
StatePublished - Aug 2010

ASJC Scopus subject areas

  • Physiology
  • Agronomy and Crop Science

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    Nguyen, H. P., Chakravarthy, S., Velásquez, A. C., McLane, H. L., Zeng, L., Nakayashiki, H., Park, D. H., Collmer, A., & Martin, G. B. (2010). Methods to study PAMP-triggered immunity using tomato and nicotiana benthamiana. Molecular Plant-Microbe Interactions, 23(8), 991-999. https://doi.org/10.1094/MPMI-23-8-0991