Wheat Protoplast Preparation and Transformation v1

Abstract

Wheat protoplast protocol for studying effector recognition and cell-autonomous defense signaling. Plant resistance (R) proteins recognize transiently expressed Avirulence (Avr) effector gene products, triggering defense responses. A co-transfected reporter plasmid enables quantification of response, upon activation of a defense-related promoter and expression of a luciferase reporter cassette. This protocol contains instructions for plant growth, protoplast isolation, transfection using the PEG-Ca method, luminescence measurement via plate reader and data processing.