LeEIX2 Interactors' Analysis and EIX-Mediated Responses Measurement

Methods Mol Biol. 2017:1578:167-172. doi: 10.1007/978-1-4939-6859-6_13.

Abstract

Plant-pathogen interactions involve a large number of wide regulatory systems, necessary for plant defense responses against pathogen attack. The fungal protein ethylene-inducing xylanase (EIX) elicits defense responses in specific cultivars of tobacco and tomato. The response to EIX is controlled by a single locus encoding for LeEIX2, a leucine-rich-repeat receptor-like-protein (LRR-RLP). As an RLP, LeEIX2 does not possess an obvious cytoplasmic signaling moiety such as a kinase domain. To study LeEIX2 mode of action, it is essential to identify the potential interactors involved after EIX perception. Here, we describe the in vivo co-IP methodology used for protein interaction verification and ethylene and ROS (reactive oxygen species) measurements used for physiological effects assessment.

Keywords: Co-IP; Defense responses; Ethylene-inducing xylanase; LRR-RLP; MAMP; ROS.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Ethylenes / analysis
  • Fungal Proteins / immunology*
  • Fungal Proteins / metabolism
  • Immunoprecipitation
  • Nicotiana / immunology*
  • Nicotiana / metabolism
  • Plant Immunity
  • Plant Proteins / metabolism*
  • Protein Binding
  • Protein Domains
  • Reactive Oxygen Species / analysis
  • Signal Transduction
  • Solanum lycopersicum / immunology*
  • Solanum lycopersicum / metabolism

Substances

  • Ethylenes
  • Fungal Proteins
  • Plant Proteins
  • Reactive Oxygen Species
  • ethylene