Glucose-6-phosphate dehydrogenase activity in spinach as measured by image analysis: a new approach for plant enzyme histochemistry.

A relatively low-cost computer-assisted image analysis system is described. Software has been specifically written for the continuous monitoring of absorbance readings on cryostat sections of plant tissues incubated in media to reveal enzyme activities. The equipment was tested by quantifying glucose-6-phosphate dehydrogenase activity in cryostat sections from shoot apices of spinach plants. The reaction rate of the dehydrogenase activity was monitored at two incubation temperatures, 20 degrees C and 30 degrees C. Control incubations were performed in media lacking substrate. The specific test minus control reaction at 30 degrees C was twice that at 20 degrees C. Variation of the substrate concentration at 30 degrees C yielded a Km value of 0.37 mM. These preliminary results show that our image analysis system can be used for kinetic measurements of dehydrogenase activity in frozen tissue sections and constitute a new approach for enzyme histochemistry in the shoot apical meristem.