Rapid method for the detection and quantification of Botrytis cinerea in plant tissues

摘要

Monoclonal antibody to B. cinerea (BC-58) was used to develop a plate-trapped antigen enzyme linked immunosorbent assay (PTA-ELISA) to detect and quantify Botrytis antigens in boysenberry flowers. The ability of antibody BC-58 to detect B. cinerea inextracts from artificially infected boysenberry flowers was assessed. Results showed that the antigen could be detected in latent infections. Antibody BC-58 sensitivity to heat treatment of the antigen, incubation conditions, and the detection limit were also investigated. Autoclaving at 121 deg C reduced the sensitivity of the antibody. Additionally, the incubation of the antigen at 4 deg C overnight produced higher absorbance values at 405 nm than incubation at 37 deg C for 2 h. The detection and quantification of B. cinerea antigen was reliable within 0-16 mu g dried mycelium per ml of PBS buffer and at least 8 X 10~3 spores.