Pigment Degradation in Discs of the Thermophilic Cucumis sativus as Affected by Light, Temperature, Sugar Application and Inhibitors

Chlorophyll degradation in Cucumis leaf discs was measured at different temperatures between 1 and 25°C in the light and in darkness, and in the presence or absence of sucrose. Two different processes of chlorophyll degradation could be distinguished, a light-requiring process operating at 1 and 5°C and another, light and sucrose enhanced degradation process which was evident at 25°C. Degradation of leaf pigments at low temperatures was of a photo-oxidative nature since there was no degradation in the dark. The chlorophyll a/b ratio was decreased, carotene was degraded at a faster rate than chlorophyll, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and triphenyltetrazolium chloride (TTC) which prevent photo-oxidation, protected against chlorophyll degradation. The light and sucrose enhanced chlorophyll degradation at 25°C was of an enzymatic nature since it occurred in the dark as well as in the light. The chlorophyll a/b ratio was not affected, and carotene and chlorophyll degradation occurred at the same rate. Since DCMU completely inhibited the light enhancement at 25°C and experimentation in a low oxygen atmosphere also protected chlorophyll against the effect of light and sugar application, it is suggested that the enhancement of chlorophyll degradation by light and sucrose at 25°C may be due to increased sugar uptake of the chloroplasts and consequently excessive starch formation in the organelles.     

Photo-oxidative Degradation of Chlorophyll-a and Unsaturated Lipids in Liposomal Dispersions at Low-Temperature

Liposomal dispersions in water were used as a tool to study photo-oxidation of chlorophyll-a and photo-oxidation of unsaturated lipids at 1 or 4°C. The presence of monogalactosyl diglyceride stimulated chlorophyll-a degradation. In addition the level of linolenic acid was decreased in liposomal dispersions containing chlorophyll-a, dipalmitoyl phosphatidyl choline, and monogalactosyl diglyceride, indicating that monogalactosyl diglyceride and chlorophyll-a were coupled in the preparations. In liposomal dispersions containing equal (molar) quantities of a-tocopherol, monogalactosyl diglyceride, and chlorophyll-a, a-tocopherol fully protected linolenic acid against photo-oxidative degradation, while chlorophyll-a degradation was only slightly reduced. In liposomal preparations containing a-tocopherol, chlorophyll-a and phosphatidyl choline, a-tocopherol catalyzed degradation of chlorophyll-a. Absorption spectra of the liposomal dispersions showed that the presence of a-tocopherol caused increased absorption in red light, which was attributed to structural changes in the liposomal preparations and thus could explain the noted effects. Tocopherol itself was rapidly degraded in chlorophyll-a containing liposomal preparations. Complex formation between chlorophyll-a and monogalactosyl diglyceride in chloroplasts is suggested and protection by a-tocopherol against photo-oxidation in chilling-sensitive plants; a suggestion which is founded on the similarities that exist between chloroplast preparations and liposomal preparations containing chlorophyll-a and monogalactosyl diglyceride as regards photo-oxidative degradation of chlorophyll-a, a-tocopherol and linolenic acid.     

Effect of α-Tocopherol, β-Carotene, Monogalactosyldiglyceride and Phosphatidylcholine on Light-Induced Degradation of Chlorophyll a in Acetone

The effect of α-tocopherol, β-carotene, monogalactosyldi-glyceride and phosphatidylcholine on red light induced degradation of chlorophyll a was studied in acetone at 4°C. Monogalaclosyldi-glyceride was ineffective up to a molar ratio of monogalactosyldi glyceride to chlorophyll of 1:10. α-Tocopherol, β-carotene and phosphatidylcholine inhibited chlorophyll degradation. Maximal protection by α tocopherol and β-carotene was similar (76%) but on a molar basis a tocopherol was less effective. Protection by phosphatidylcholine was less than by a tocopherol and α-carotene but the lipid was effective at a lower ratio of chlorophyll to protectant. Inhibition by phosphatidylcholine was independent of the degree of unsaturation of the fatty acids. Effects of β-carotene and α-tocopherol were additive at suboptimal concentrations, but addition did not increase the maximal protection of 76% by these substances alone. Phosphatidylcholine increased the effectiveness of α-tocopherol and β-carotene independent of their concentrations. It is suggested that interactions between lipids participate in the mechanism protecting chlorophyll a against photooxidation in the chloroplast membrane.     

Photooxidative damage to the photosynthetic apparatus during chilling

Short periods of chilling in the presence of light (up to 6 h: 1°C; 270 W/m²) decreased the subsequent apparent photosynthesis and chlorophyll fluorescence of leaf discs of Cucumis sativus L. cv. Kleine Groene Scherpe. The extent of the injury depended on the duration of the chilling pretreatment. After 6 h the subsequent apparent photosynthesis even reached a negative value, and it increased only slightly during the next 2 1/2 h. The decrease of apparent photosynthesis was not a consequence of increased dark respiration but was of photooxidative origin since the presence of both light and oxygen was required. Preincubation in the light for 2 h at 20°C sensitized leaf discs to subsequent photooxidation during chilling. Prompt and delayed chlorophyll fluorescence decreased simultaneously after chilling and light treatment. The corresponding decrease of photosynthesis and chlorophyll fluorescence is discussed in relation to primary photooxidative damage to the photosystems in the chloroplast membrane.