Combining Clearing and Fluorescence Microscopy for Visualising Changes in Gene Expression and Physiological Responses to Plasmodiophora brassicaeDeeksha Singh 1, Sara Blicharz 1, Karolina Stefanowicz 1, Laura Ragni 2, Kornel Michalak 3, Agnieszka Bagniewska-Zadworna 3, Robert Malinowski 1
1Integrative Plant Biology Team, Institute of Plant Genetics Polish Academy of Sciences, 2ZMBP-Center for Plant Molecular Biology, University of Tübingen, 3Department of General Botany, Institute of Experimental Biology, Faculty of Biology, Adam Mickiewicz University
The present protocol describes an optimized method for the histological observation of galls induced by Plasmodiophora brassicae. Vibratome sections of hypocotyls are cleared before fluorescence imaging to study the involvement of transcription factors and phytohormones during disease progression. This protocol overcomes resin embedding limitations, enabling in planta visualization of fluorescent proteins.
