An investigation has been made of the combined effects of low temperature and high light on the level of several photosynthetic products in the leaves of a group of plants differing widely in their tolerance to this stress. Starch levels in these plants after chilling are dependent on the time of day that temperatures are lowered and seem related to rates of C02 assimilation under this stress. Prolonged low-temperature, highlight treatment (10 C at 160 wmr') of Sorghum bicolor induced a rapid starch hydrolysis after a lag of some 24 hours. Differing rates of starch loss at the cellular level and a rapid migration of chloroplasts toward the base of upper mesophyll cells were also seen in legves of this stress-sensitive species.Chilling increased the level of almost all free amino acids in tolerant and in semi-tolerant species, while amino acids related to intermediates of the C4-pathway show a sharp or transitory decrease in Sorghum. These and other changes observed in Sorghum suggest that some time-and temperature-dependent blockages develop in the interconversion of C4-pathway intermediates and possibly in the flow of other intermediates to and from the sites of C4-photosynthesis.Levels of ATP in the leaves of Sorghum, Paspalum, and Amaranthus increased at night and following chilling and did not fall until pronounced necrosis of the leaves commenced.Previous reports (21,22) have shown that high light levels combined with low temperatures can cause necrotic lesions to develop on the leaves of several plants. Lower levels of light prevent or slow the onset of these lesions. Some species of both C3-and C4-photosynthetic pathway (7) Lolium multiflorum L. var Grasslands "Tama" Westerwolds.Potting media, plant nutrients, and controlled environment cabinets were as described previously (21). Plants were given three phases of treatment. They were first preconditioned at a constant day-night temperature of 25 C for 11 to 14 days. Temperatures were generally lowered at the start of a photoperiod to a constant 10 C day and night for 3 days and then returned to 25 C. Photoperiods were 12 hr with light intensities maintained at 160 wm-2 (400-700 nm).Soluble Sugar, Starch, and Malate Assays. Several leaves with most recently emerged ligules (youngest-mature) were harvested from different plants 3 hr before the completion of 12-hr photoperiods. Leaves were lyophilized and ground, and aliquots were used for subsequent analyses. Soluble sugars extracted in 80% (v/v) ethanol and starch extracted from the alcohol-insoluble residue according to Hassid and Neufield (6) were assayed with phenol-sulphuric acid (1). Malate was extracted from aliquots (250 mg) of the same dry leaf powder in 5 ml of 6% HCIO4 at 0 C and determined enzymically (8).Similar assays were also used to determine the levels of leaf starch at more frequent intervals before and during low-temperature treatment.Amino Acid Assays. Amino acids were extracted from 1.5 g wet weight of youngest-mature leaf lamina in methanol-chloroform (2:1) followed by 0.1 ...