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Formation of major prenylquinones and carotenoids was investigated by comparing the incorporation of [14C]mevalonate into segments of different age from green and etiolated leaves of 22 C-grown rye seedlings (Secale cereale L.) and from 32 C-grown rye leaves which contained bleached and proplastid-like ribosome-deficient plastids, due to a heat-sensitivity of 70S ribosome formation. The contents of plastidic isoprenoids were much lower (between 2 - 30%) in the achlorophyllous than in green leaves. In green leaves [14C]mevalonate incorporation into non-polar lipids and into plastoquinone was partially inhibited in the presence of gabaculin, an inhibitor of chlorophyll synthesis. However, except for β-carotene, [14C]mevalonate incorporation into isoprenoids continuously increased with age also in achlorophyllous etiolated or 32 °C-grown, as in green, leaves and was, except for P-carotene and plastoquinone, higher in etiolated than in green leaves. In bleached °32 C-grown leaves [14C]mevalonate incorporation into all plastidic isoprenoids was strikingly (up to 45-fold) higher than in green control leaves. While degradation of P-carotene was greatly enhanced in bleached 32 °C-grown leaves, relative to green control leaves, and could thus compensate for a higher apparent synthesis, chase experiments did not reveal any marked differences of the turnover of other isoprenoids. The half times of plastoquinone. phylloquinone and lutein were in the order of 2-3 days. Within a 24 h chase period a-tocopherol degradation did not become apparent. Uptake of [14C]mevalonate and [14C]isopentenyl pyrophosphate by isolated bleached plastids from 32 °C-grown leaves was much more rapid than by chloroplasts and resulted in higher precursor accumulation within the organelle. While mevalonate incorporation into isoprenoid lipids was not detected, isopentenyl pyrophosphate was incorporated into isoprenoid lipids, including plastoquinone. Rates of incorporation by isolated chloroplasts or bleached plastids were of similar order. The results illustrate that divergent types of plastid differentiation are associated with fundamental developmental changes of the metabolic flow of isoprenoid precursors between different products and compartments and, in particular, with changes of import into the plastid compartment.
Among chlorosis-inducing herbicides that interfere with carotenoid synthesis two groups of different potency can be discriminated (group 1: aminotriazole amd haloxidine; group 2 with more extensive photodestructions: pyridazinone herbicides and difunon). After application of herbicides of group 2 colored carotenoids were completely absent and preexisting chlorophyll was degraded by photochemical reactions requiring high light intensity and O2, that occurred also at 0°C. In treatments with group 1 herbicides direct photodegradation of chlorophyll was not sufficient to generate the chlorosis. Light-induced interference with constituents of the chloroplast protein synthesis apparatus being more sensitive to photooxidative damage than chlorophyll, appeared to indirectly mediate the chlorosis. In the absence of chloroplast protein synthesis further chlorophyll accumulation is prevented. Photodegradation of chlorophyll in the presence of group 2 herbicides involved the participation of O2- radicals and was accompanied by lipid peroxidation. In all herbicide treatments the catalase activity of the leaves was very low. Only in the presence of group 2 herbicides chloroplast enzymes of cytoplasmic origin (e.g. NADP-glyceraldehyde-3-phosphate dehydrogenase) were also inactivated. Rapid inactivation of catalase as well as of NADP-glyceraldehyde-3-phosphate dehydrogenase was induced by exposure of dim-light-grown herbicide-treated leaves to bright light, also at 0°C. In treatments with herbicides of group 2 also other peroxisomal enzymes (e.g. glycolate oxidate, hydroxy-pyruvate reductase) were affected. The elimination of these peroxisomal enzymes also appeared to depend on photooxidative processes of the chloroplast.
Synechococcus (Anacystis nidulans, strain L 1402-1) were grown at + 37 °C in an atmosphere of 0.04 vol.% CO2 using different light conditions. Changing the culture conditions caused alterations in pigment ratios and ultrastructure of Synechococcus. In comparison to the low white and red light grown cells under strong white light the number of thylakoids decreased and an accumulation of storage carbohydrates could be observed. The number of the polyhedral bodies also varied with culture conditions. The results are discussed with reference to the pigment composition and the function of the polyhedral bodies.
The cyanobacteria Anabaena cylindrica and Synechococcus leopoliensis (= Anacystis nidulans) were grown at different levels of UV-B radiation (439. 717, 1230 and 1405 J m -2d-1 weighted according Caldwell, 1971) for 2 days. Dry weight was hardly affected but phycocyanin content of both species decreased linearly to the level of UV-B radiation. Contents of protein, carotenoids and chlorophyll a were reduced only after exposure to high doses (1230 J m-2d-1) of UV-B radiation. Photosynthetic 14CO2 fixation of Anabaena cells was reduced linearly with increasing UV-B dose whereas no effect could be observed in Synechococcus. A depression of photosynthetic 15N-nitrate uptake was found after UV-B stress in both species. UV-B irradiance caused an increase of 15N-incorporation into glutamine, but no effect was noted for incorporation into alanine or aspartic acid. An increase of 15N-excess in glutamic acid linear with the UV-B dose was observed in Synechococcus, only. Patterns of 14C-labelled photosynthetic products were either less affected by UV-B radiation (Anabaena) or an enhancement of 14C-label in total amino acids was detected (Synechococcus). The amount of total free amino acids increased parallel to the level of UV-B radiation. Only, the high dose of UV-B (1405 J m-2d-1, weighted) results in a decrease of the glutamine pool. Our results indicate an inhibition of glutamate synthase by UV-B irradiation in Anabaena, only. Results were discussed with reference to the damage of the photosynthetic apparatus.
The marine diatoms Bellerochea yucatanensis and Thalassiosira rotula were grown at different salinities (20/25, 35, and 40/45‰ salinity (S), respectively) under normal air (0.035 vol.% CO2). No significant variations in the percentage of gross photosynthetic products (e.g. total amino acids, sugar phosphates) were found as a function of salinity during growth. The bulk of the soluble 14C-radioactivity was detected in amino acids. 14C-labelling of glutamine increased markedly with salinity. Low salt - grown algae are characterized by enhanced amino acid pools, mainly of aspartic acid, asparagine and glutamine. It was found that the tested amino acids are not involved in osmoregulation.
The cyanobacterium Synechococcus (Anacystis nidulans, strain L 1401-1) grown under different light conditions showed variations in pigmentation. Ratios of photosynthetic pigments and the effect on quantum requirement and oxygen evolution were studied. An increase in the ratio of chlorophyll a forms with absorption maxima in the far red regime to total chlorophyll a forms was observed in cells grown in strong white light. The quantum efficiency of orange light (637 nm) - absorbed by phycocyanin - was higher after growth of Synechococcus in white than in red light. The quantum efficiency at 677 nm increased when cells were grown in red light and decreased strongly after transfering red light grown cells to conditions of strong white light. The results show an adaptation of pigment composition to light regimes during growth and its effect on photosynthesis.
14C-and 15N-Assimilation, 15N-Labelled Amino Acids, M arine D iatom s The marine diatoms Bellerochea yucatanensis and Skeletonema costatum were grown at +20 °C in 0.03 vol.% CO2 with nitrate or ammonia. The 15N -am m onia and 15N -nitrate assim ila tion and 15N -incorporation into various amino acids were studied of both diatom s during exponential growth phase in dependence of different nitrogen conditions. In all experiments the 15N -am m onia uptake was lower than the 15N -nitrate assim ilation rate up to 20-40 min photo synthesis. N itrate lim itation -cells grown in nitrate followed by growth in nitrogen-free m edium for 24 h — caused a strong 15N-label into aspartate after adding 15NH 4C1 (1 m M). In cells grown in nitrate highest enrichment of 15N was found in glutamine. Results were discussed with reference to the operating of the GS/GOGAT system and glutam ic acid dehydrogenase pathway. Photosynthetic 14CO2 fixation experiments showed a very high labelling of aspartate which was interpreted with a phosphoenolpyruvate carboxylation catalysed by phosphoenolpyruvate carb-oxykinase.
The marine diatom Ditylum brightwellii (West) Grunow isolated from the Baltic Sea could be synchronized by a light/dark rhythm of 6.5:17.5 h (white light intensity 8 W m-2) at 18 °C and 0.035 vol.% CO2. Content of protein, DNA and RNA increased linearly up to the end of the cell cycle. Pigments (chlorophyll a, chlorophyll c1 + c2, carotenoids) and galactolipids were synthesized in the light period only. A lag phase of 2 h was observed in the biosynthesis of sulphoquinovosyl diacylglycerol and phosphatidylglycerol. Formation of phosphatidylglycerol and phosphatidylcholin continued in the dark period (30% and 28%, respectively). The pattern of major fatty acids (C14:0, C16:1, C16:0, C18:1 and C20:5) varied during the cell cycle of Ditylum.
Biosynthesis of acyl lipids was reduced in dependence on the UV-B dose. The most sensitive lipid was digalactosyl diacylglycerol (total inhibition at 585 J m-2), whereas phosphatidylcholin was less affected (20% reduction). UV-B radiation during the dark period had no effect on the lipid and pigment content. Strongest inhibitory effect of UV-B on cell division, synthesis of protein, pigments, sulphoquinovosyl diacylglycerol and phosphatidylglycerol was found after UV-B radiation at the beginning of the cell cycle (0.-2. h). An exposure time at the end of the light period (4.-6. h) led to a marked damage on the synthesis of monogalactosyl diacylglycerol and phosphatidylglycerol. These findings indicate a stage-dependent response of Ditylum to UV-B irradiance. The impact of UV-B resulted in an increase of unsaturated long chained fatty acids (C18, C20) and in a diminution of short chained fatty acids (C14, C16). Content of ATP was not affected by UV-B radiation under the used conditions. The inhibitory effect of UV-B on synthesis of DNA, RNA, protein and acyl lipids was mainly reversible. Results were discussed with reference to UV-B damage on the enzymes involved in the biosynthesis of acyl lipids and by a reduction of available metabolites.
The cyanobacterium Synechococcus leopoliensis (Anacystis nidulans, strain L 1402-1) grown at 39 °C and 2 vol. % CO : could be synchronized by a light/dark regime of 3:5 h (white light intensity 1.5 × 104 erg cm-2 sec-1). Content of pigments (chlorophyll a. phycocyanin and carotenoids), R N A and proteins increased linearly up to 100% at the end of the light period while DNA synthesis was lower. Chlorophyll a synthesis was correlated to the photosystem I activity of the isolated thylakoids and to the formation of MGD G . Galacto lipids were synthesized in the light period, only. A lag phase of 2h was observed in the biosynthesis of SQDG and PG. No significant differences were found between the cell and thylakoid fractions. Palmitic (C16:0), hexadecenoic (C16:1) and octadecenoic (C18:1) acid as major com ponents accounted for more than 90% of total fatty acids in MGD G , DGDG and SQDG . PG contains a small amount of stearic (C18:0) and heptadecenoic (C17:1) acid. No significant variations in the fatty acid distribution of all lipids could be detected in the cell fraction during the division cycle. Changes in the ratio of saturated to unsaturated fatty acids were found in isolated thylakoids. only. In experiments with [14C]bicarbonate main radioactivity was measured in galacto lipids while using [14C]acetate SQDG and PG were markedly [14C]labelled. Results were discussed with reference to the findings of eucaryotic algae and the formation of photosynthetic membranes.
The growth of Synechococcus at different intensities of white and red light caused changes in the pigment composition. The ratio of chlorophyll a to phycocyanin varied from 1:8,2 in LWLI-grown cells to 1:1,4 in cells grown at HWLI and to 1:15,7 in cultures exposed to HRLI. Acyl lipids were quantitatively determ ined and fatty acids of the individual lipid classes analysed by GLC. Phycocyanin-free photosynthetic lam ellae were obtained by fractional centrifugation. No variation was found in the acyl lipid composition of the m em brane preparations. These all contained MGDG, DGDG, SQDG and PG as components. In all the lipids investigated, palmitic, hexadecenoic and octadecenoic acids m ade up to more than 90% of total fatty acids. The pattern of these major components w ithin the lipids from the different cultures depended on the light used. No large differences were detected between zones obtained from LWLI and HRLI isolated membranes, whereas density gradient centrifugation of those from HWLI-grown cells resulted in a completely different pattern of bands. The variations in lipid and fatty acid composition are discussed with respect to changes observed in lipid composition of whole cells and the results reported on tem perature dependent shifts in lipid fluidity in cyanobacteria.