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Methods to Study PAMP-Triggered Immunity Using Tomato and Nicotiana benthamiana

August 2010 , Volume 23 , Number  8
Pages  991 - 999

Hanh P. Nguyen,1,2 Suma Chakravarthy,1,3 André C. Velásquez,1,3 Heather L. McLane,1 Lirong Zeng,1,4 Hitoshi Nakayashiki,5 Duck-Hwan Park,3 Alan Collmer,3 and Gregory B. Martin1,3

1Boyce Thompson Institute for Plant Research, Ithaca, NY, U.S.A.; 2Department of Plant Biology, Cornell University, Ithaca, NY, U.S.A.; 3Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, NY, U.S.A.; 4Biology Department, University of Arkansas at Little Rock, Little Rock, AR, U.S.A.; 5Laboratory of Plant Pathology, Faculty of Agriculture. Kobe University, 1-1, Rokkodai-cho, Nadu-ku, Kobe 657-8501, Japan

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Accepted 30 March 2010.

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 pathosystems, such as those involving Solanaceae plant--pathogen 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 mitogen-activated 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.

© 2010 The American Phytopathological Society