AgroLux: bioluminescent Agrobacterium to improve molecular pharming and study plant immunity

• DOI: 10.1111/tpj.15454
• Publication Date: 2021-08-09T08:58:41Z
• Authors: Philippe V. Jutras; Riccardo Soldan; Isobel Dodds; Mariana Schuster; Gail M. Preston; Renier A. L. van ...
• Keywords: Nicotiana; 0301 basic medicine; 0303 health sciences; Light; Green Fluorescent Proteins; Molecular Farming; Recombinant Proteins; Fungal Proteins; Plant Leaves; 03 medical and health sciences; Agrobacterium tumefaciens; Luminescent Measurements; Operon; Plant Immunity; Microorganisms, Genetically-Modified

SUMMARYAgroinfiltration in Nicotiana benthamiana is widely used to transiently express heterologous proteins in plants. However, the state of Agrobacterium itself is not well studied in agroinfiltrated tissues, despite frequent studies of immunity genes conducted through agroinfiltration. Here, we generated a bioluminescent strain of Agrobacterium tumefaciens GV3101 to monitor the luminescence of Agrobacterium during agroinfiltration. By integrating a single copy of the lux operon into the genome, we generated a stable ‘AgroLux’ strain, which is bioluminescent without affecting Agrobacterium growth in vitro and in planta. To illustrate its versatility, we used AgroLux to demonstrate that high light intensity post infiltration suppresses both Agrobacterium luminescence and protein expression. We also discovered that AgroLux can detect Avr/Cf‐induced immune responses before tissue collapse, establishing a robust and rapid quantitative assay for the hypersensitive response (HR). Thus, AgroLux provides a non‐destructive, versatile and easy‐to‐use imaging tool to monitor both Agrobacterium and plant responses.