A Rapid Microtitre Plate Assay for Determining Sensitivity to Photosystem II Herbicides

Abstract

This paper reports on a microtitre plate version of the 2,6-dichlorophenol-indophenol (DCPIP) reduction assay. DCPIP reduction rates of alfalfa thylakoid membranes were determined by measuring absorbance at 600 nm before and after a 1 min illumination period. The membranes were near light-saturated at intensities above 600 μE m–2s–1. Saturation of thylakoid membranes with DCPIP occurred above 120 μM. DCPIP reduction rates increased linearly with chlorophyll concentrations from 0.25 to 2 μg. The rate of DCPIP reduction was linear throughout a 2 min illumination period (R2= 0.99). The assay was sensitive enough to terbacil to differentiate between a 20% change in the I50concentration. DCPIP reduction rates were sensitive to concentrations of terbacil as low as 100 nM. It only takes 13 minutes to load and read 96 samples using the microtitre plate assay compared to 5.5 h using the conventional procedure.