The effect of leaf detachment on chlorophyll fluorescence was analyzed for Zea mays, Cucumis sativus, Phaseolus vulgaris, and Echinochloa crus-galli. Results clearly indicate that detachment hastens the decrease in chlorophyll fluorescence during the course of chilling experiments. For maize and bean, the activity of photosystem II of chloroplasts isolated from detached leaves is lower than that of chloroplasts isolated from attached leaves. There are also large differences in ionic loss between detached and attached leaves of barnyard grass which could correlate with changes in leaf water status. The detached leaves lost some 50% of their total ionic content. Finally, detachment alters the ranking of the species with regard to their chilling tolerance.There has been much interest in the use of room temperature Chl fluorescence as a probe for chilling injury (2,3,(8)(9)(10). In the procedure developed by Smillie and coworkers (8-10) assess chilling tolerance in plants, fluorescence is measured on detached leaves stored at 0°C in the dark. Changes in the Chl fluorescence induction curve are followed on the same excised leaves for time periods ranging from several h to weeks (9). Such chilling in the dark results in a decrease in the slope of the rising phase of the Chl fluorescence induction (8) which has been interpreted as a reflection of damage occurring on the photooxidizing side of PSII (6). However, it has been shown, using isolated chloroplasts of bean and tomato, that detachment and dark storage cause more damage to PSII than does the cold treatment itself (4). Before using changes in Chl fluorescence of detached leaves as an indication of chilling tolerance, one must be assured that detachment itself does not affect the 'fluorescence as it affects isolated chloroplasts. The present study was therefore carried out to determine the effects of detachment of the leaves on Chl fluorescence during chilling experiments. 'hybrid GH-390'; Phaseolus vulgaris (L.) cv 'Canadian Wonder'; and Cucumis sativus (L.) cv 'Palomar'. Seeds were sown in a mixture of 2: 1:1 perlite, vermiculite, and peat moss and grown during June and July in Sydney, Australia, in a temperaturecontrolled greenhouse with a minimum of 17°C and a maximum of 28°C. For maize and barnyard grass, light was supplemented to yield 600 gmol -m-2. s-' at noon, by two incandescent bulbs (40 w) and six fluorescent lamps (80 w) placed 70 cm above the plants. Daylength was set at 14 h. Plants were watered daily with 6% Hoagland nutrient solution. Plants were used at age 6 to 7 weeks for barnyard grass, 4 to 5 weeks for maize, 4 weeks for cucumber, and 3 weeks for bean. Each experiment was replicated twice with a new group of plants and, in the case of barnyard grass, plants used in the replicate experiment were sown 6 weeks after the first cohort to rule out either photoperiod or greenhouse effects. All the results were statistically tested by analysis of variance and Tukey's honestly significant differences a posteriori method (11).
MATERIALS AND MET...