A comparison study of detection systems for plant innate immune responses with tobacco BY-2 suspension cells

• 1、Plant Pathology, College of Agriculture and Biotechnology, China Agricultural University, Bei Jing 100193

Abstract：Isolation and identification of pathogen elicitors that induce plant immune responses is crucial to research on mechanisms of plant innate immunity. To establish a system suitable for large scale screening, we have investigated 3 assay systems of tobacco BY-2 cell suspensions in response to diverse pathogen-associated molecular patterns (PAMPs) using chlorophenol red, luminol and 5-aminosalicylic acid as detection substrate respectively. Results showed that the chlorophenol red system worked well under lower cell densities, and the reaction was swift and macroscopic, but this system may not suit screening assays of proteinaceous elicitors due to buffers commonly used during extraction processes. The enzyme-mediated luminol chemiluminescence system was vulnerable to a variety of factors and unfit for the analysis of crude protein extracts as well. In contrast, the 5-aminosalicylic acid detection system worked better at higher cell densities, and the color reaction was fast, stable and intensified over time. In addition to these advantages, the cost of 5-aminosalicylic acid detection is low, which collectively makes it suitable for the high-throughput screening. Thus, the established 5-aminosalicylic acid detection system lays the foundation for further functional genomic exploration of pathogen elicitors in future.

Keywords： plant pathology tobacco cell suspensions BY-2 plant innate immunity PAMPs elicitors