A novel open-system technique to monitor real-time oxygen consumption during early phases of seed germination

Abstract

A novel technique allows long-term monitoring of real-time oxygen consumption during seed germination in an open system. Most current techniques used to detect oxygen consumption by seeds measure the decrease in oxygen concentration in a closed chamber. This is not ideal for long-term experiments because the chamber must be replenished with air periodically, subjecting the seeds to abrupt changes in oxygen concentration. The current technique employs an open system, in which seeds are submerged in a continuously aerated aqueous environment. Oxygen electrodes are used to measure the steady-state concentration of oxygen in the solution, which is a function of both the rate of oxygen consumption by the seed and the rate of aeration from the atmosphere. The rate of aeration is directly dependent on the oxygen concentration of the bathing solution; therefore, previous calibration of the system allows the direct conversion of steady-state oxygen concentrations into oxygen consumption rates. Because oxygen is not limiting, the experimental design described here can monitor the same sample non-intrusively every minute for more than 24 h, allowing for greater precision than hourly readings often reported with current techniques. Multiple treatments and/or replicates can be run simultaneously, allowing sensitive comparison of various seed treatments or seed types. To illustrate its potential application, the technique was used to follow the rehydration and pre-emergence phases of germination of cucumber (Cucumis sativum), pea (Pisum sativum) and mustard (Brassica juncea) seeds, detect the inhibitory effects of surface sterilization techniques on seed respiration of cucumber, and follow the interaction of a bacterial biocontrol agent with germinating cucumber and pea seeds.