Studies into the Action of Some Photosynthetic Inhibitor Herbicides

Treatment of flax cotyledons with solutions of bromacil, ioxynil,metribuzin, and monuron resulted in a rapid inhibition of CO₂fixation. Chloroplasts isolated from cotyledons treated withthe above herbicides exhibited electron transport activity forsome time after CO₂ uptake in the intact cotyledons had ceased.However, the action of the herbicides was to decrease photosystemtwo (PSII) activity faster than photosystem one (PSI) activitybut there appeared to be no differential breakdown of the pigmentcomplexes resolved by polyacrylamide gel electrophoresis. Carotenoidbreakdown, particularly - and ß- carotene, precededchlorophyll loss following herbicide treatment. Ethane generation,an indicator of membrane breakdown, increased as the pigmentlevels decreased. Incubation of cotyledons in an argon atmospherereduced the rate of pigment loss with ioxynil treatment andprevented pigment breakdown with monuron treatment. Pigmentloss was also reduced by the addition of a singlet oxygen quencher,l, 4-diazo-bicyclo(2, 2, 2)octane (DABCO). Ultrastructural effectscommenced with a rapid swelling of the chloroplast followedby a gradual disorganization and swelling of the thylakoid systemand subsequent tonoplast rupture. With ioxynil numerous membrane-boundvesicles were formed in the chloroplast prior to envelope rupture.The evidence suggests that, following inhibition of electrontransport, the photosynthetic inhibitor herbicides exert theirtoxic effect by causing the formation of excited singlet oxygen.It is suggested that this is initially quenched by the chloroplastprotective mechanisms; however, since these systems are overloadedsubsequent membrane breakdown occurs.     

Role of Nitrate in Photosynthetic Electron Transport of Chlorella Vulgaris

Addition of nitrate to a suspension of NO₃ ^--depleted Chlorella vulgaris cells raised the O₂-evolving capacity of the organism by 60%. The rate of O₂-evolution under flash irradiation of the depleted cells was drastically reduced, which could be restored by addition of NO₃ ^-. The 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB)-insensitive O₂-evolution, i.e., photosystem (PS) 2 activity of NO₃--depleted cells, showed a 75% stimulation by addition of NO₃ ^-. PS1-mediated electron transport was also stimulated (50%) by addition of NO₃ ^-. Fluorescence yields of the NO₃ ^--depleted cells were significantly reduced. A normal fluorescence response was restored by the addition of NO₃ ^-. The fluorescence yield of the NO₃ ^--depleted and DCMU-treated-cells increased significantly after addition of NO₃ ^- ions, indicating a further reduction of the primary acceptor of PS2 (Q). In addition, the low temperature fluorescence emission spectra showed that energy transfer to PS2 and PS1 was much higher when nitrate was present. Hence nitrate accelerates the light-induced charge transfer from the intact O₂-evolving system to the primary electron acceptor of PS2 and stimulates the PS1-mediated electron transport. This revised version was published online in June 2006 with corrections to the Cover Date.     

Long-term delayed luminescence in Scenedesmus obliquus. II. Influence of exogeneous factors

Long-term delayed luminescence varying from 0.3 s up to several minutes has been studied in wild type and several pigment mutants of Scenedesmus obliquus during the life cycle and under the influence of various exogeneous parameters such as herbicides, different pH values, temperature, preillumination, and the diurnal rhythm of synchronized cells. All these parameters investigated exhibit a specific and distinct impact. Therefore, long-term delayed luminescence may serve as some kind of assay of the ‘status of vitality’ of green living cells, i.e., as a fast and simple screening procedure of potentially harmful environmental factors, and also for herbicides. In addition, only intact chloroplasts show long-term delayed luminescence suggesting a possible assay for the physiological activity during isolation of chloroplasts.     

The nucleotide sequence of Scenedesmus obliquus chloroplast tRNAfMet.

The chloroplast initiator tRNAfMet from the green alga Scenedesmus obliquus has been purified and its sequence shown to be p C-G-C-A-G-G-A-U-A-G-A-G-C-A-G-U-C-U-Gm-G-D-A-G-C-U-C-m2(2)G-psi-G-G-G-G-C-U-C-A -U-A-A-psi-C-C-C-A-A-U-m7G-D-C-G-C-A-G-G-T-psi-C-A-A-A-U-C-C-U-G-C-U-C-C-U-G-C-A-A-C-C-A-OH. This structure is prokaryotic in character and displays close homologies with a blue green algal initiator tRNAfMet and bean chloroplast initiator tRNAfMet.     

Changes in the Reactivity of the Photosynthetic Apparatus in Heterotrophic Ageing Cultures of Scenedesmus obliquus

Ultrastructural changes of the chloroplast which occur parallel to the decay of photosynthetic capacity during the heterotrophic ageing of Scenedesmus cells have been studied. A gradual disappearance of the starch grains took place as the nutrients in the growth medium were depleted. Accompanying this change was a deterioration in the structure of the cytoplasm coupled with a concomitant granular appearance. At the same time thylakoid structures were not only well preserved but accumulated more material. This resulted in an increasing number of stacked thylakoids and a thickening of the membranes.     

Studies of Electron Transport Dynamics in Photosynthetic Reaction Centers Using Fast Temperature Changes

Rates of thermoinduced conformational transitions of reaction center (RC) complexes providing effective electron transport were studied in chromatophores and isolated RC preparations of various photosynthesizing purple bacteria using methods of fast freezing and laser-induced temperature jump. Reactions of electron transfer from the primary to secondary quinone acceptors and from the multiheme cytochrome c subunit to photoactive bacteriochlorophyll dimer were used as probes of electron transport efficiency. The thermoinduced transition of the acceptor complex to the conformational state facilitating electron transfer to the secondary quinone acceptor was studied. It was shown that neither the characteristic time of the thermoinduced transition within the temperature range 233-253 K nor the characteristic time of spontaneous decay of this state at 253 K exceeded several tens of milliseconds. In contrast to the quinone complex, the thermoinduced transition of the macromolecular RC complex to the state providing effective electron transport from the multiheme cytochrome c to the photoactive bacteriochlorophyll dimer within the temperature range 220-280 K accounts for tens of seconds. This transition is thought to be mediated by large-scale conformational dynamics of the macromolecular RC complex.     

Role of heterotrophic nutrition in growth of the alga Scenedesmus obliquus in high-rate oxidation ponds.

The green alga Scenedesmus obliquus readily adapted to heterotrophic growth in the dark, utilizing glucose as the sole carbon source. Heterotrophic cells differed significantly from photoautotrophic cells with respect to several physiological properties such as the rate of photoassimilation of CO2, rate of incorporation of glucose, and chlorophyll a concentration. Oxidation pond cells shared features common to both photoautotrophic and heterotrophic cells. Approximately 15 percent of oxidation pond algal carbon was derived from glucose assimilated directly without first being oxidized by bacteria. Bacteria seem to play a minor role in biological oxygen demand reduction in high-rate oxidation ponds, and their role is probably confined to degradation of biopolymers, thus producing substrates for algal consumption.     

Regulation of Photosynthetic Electron Transport in Intact Spinach Chloroplasts: II. MECHANISM OF SALT-INDUCED INCREASE IN OXALOACETATE PHOTOREDUCTION

The main focus of this study was to determine the mechanism by which certain exogenous monovalent salts stimulate rates of net O₂ evolution linked to oxaloacetate reduction in intact spinach chloroplasts. The influence of salts on the dicarboxylate translocator involved in the transport of oxaloacetate and on the activity and activation of the chloroplast enzyme NADP-malate dehydrogenase, which mediates electron transport to oxaloacetate, was examined. High concentrations of KCl (155 millimolar) increased the apparent K_m for oxaloacetate but did not significantly alter the maximal velocity of uptake. Likewise, external salts (KCl, MgCl₂, or KH₂PO₄) had minimal effects on the magnitude of light activation of NADP-malate dehydrogenase. In contrast, measurements of chloroplast NADP-malate dehydrogenase activity (after release by osmotic shock) showed a marked dependence on salt concentration. Rates were stimulated approximately 2-fold by both monovalent (optimally 75 millimolar) and divalent (optimally 20 millimolar) salts. It was inferred that the salt-induced increase in net rates of O₂ evolution linked to oxaloacetate reduction is due, at least in part, to stimulation of NADP-malate dehydrogenase caused by monovalent cation permeability of the chloroplast inner envelope membrane.     

Photosynthetic Acclimation to Temperature in the Desert Shrub, Larrea divaricata: II. Light-harvesting Efficiency and Electron Transport

The response of photosynthetic electron transport and light-harvesting efficiency to high temperatures was studied in the desert shrub Larrea divaricata Cav. Plants were grown at day/night temperatures of 20/15, 32/25, or 45/33 C in rough approximation of natural seasonal temperature variations. The process of acclimation to high temperatures involves an enhancement of the stability of the interactions between the light-harvesting pigments and the photosystem reaction centers. As temperature is increased, the heat-induced dissociation of these complexes results in a decrease in the quantum yield of electron transport at limiting light intensity, followed by a loss of electron transport activity at rate-saturating light intensity. The decreased quantum yield can be attributed to a block of excitation energy transfer from chlorophyll b to chlorophyll a, and changes in the distribution of the excitation energy between photosystems II and I. The block of excitation energy transfer is characterized by a loss of the effectiveness of 480 nm light (absorbed primarily by chlorophyll b) to drive protochemical processes, as well as fluorescence emission by chlorophyll b.     

Role of heterotrophic nutrition in growth of the alga Scenedesmus obliquus in high-rate oxidation ponds

The green alga Scenedesmus obliquus readily adapted to heterotrophic growth in the dark, utilizing glucose as the sole carbon source. Heterotrophic cells differed significantly from photoautotrophic cells with respect to several physiological properties such as the rate of photoassimilation of CO2, rate of incorporation of glucose, and chlorophyll a concentration. Oxidation pond cells shared features common to both photoautotrophic and heterotrophic cells. Approximately 15 percent of oxidation pond algal carbon was derived from glucose assimilated directly without first being oxidized by bacteria. Bacteria seem to play a minor role in biological oxygen demand reduction in high-rate oxidation ponds, and their role is probably confined to degradation of biopolymers, thus producing substrates for algal consumption.     

Photosynthetic Electron Transport Chain of Chlamydomonas reinhardi VI. Electron Transport in Mutant Strains Lacking Either Cytochrome 553 or Plastocyanin

A mutant strain of Chlamydomonas reinhardi, ac-206, lacks cytochrome 553, at least in an active and detectable form. Chloroplast fragments of this mutant strain are inactive in the photoreduction of NADP when the source of electrons is water, but they are active when the electron source is 2,6-dichlorophenolindophenol and ascorbate. The addition of either cytochrome 553 or plastocyanin, obtained from the wild-type strain, has no effect upon the photosynthetic activities of the mutant strain. Cells of the mutant strain lack both the soluble and insoluble forms of cytochrome 553, but they possess the mitochondrial type cytochrome c. Thus, the loss of cytochrome 553 appears to be specific.     

Photosynthetic Capacity and Quantum Requirement of Three Secondary Mutants of Scenedesmus obliquus with Deletions in Carotenoid Biosynthesis*

Photosynthetic capacity and quantum requirement of photosynthesis were evaluated in three secondary mutants of Scenedesmus obliquus possessing specific blocks in carotenoid biosynthesis. These were derived from the mutant, C-2 A', a strain which develops chlorophyll only in the light. All strains were capable of a normal and rapid production of chlorophyll and the development of a functional photosynthetic apparatus when exposed to light. LHC-II levels in the mutants lacking the β,ε-carotenoids (strains C-2 A'-34,1 and C-2 A'-67, 3b) were considerably reduced in fully greened cells but normal in the β,β-epoxycarotenoid deficient-only strain (C-2 A'-67,1) (Bishop, 1996). The maximum photosynthetic capacity and the quantum efficiency (at λ=682nm) of photosynthesis of cells of C-2A'-34,1 and C-2 A'-67,1 showed no significant changes when compared to the original parent strain, C-2 A'. However, the quantum requirement of the strain lacking both types of carotenoids, C-2 A'-67,3b, was slightly increased. It is of particular interest that both strains lacking the β,ε-carotenoids also show a preferential loss of the oligomeric form of the LHC and its associated chlorophylls. The potential requirement of the different carotenoids in maintenance of photosynthetic efficiency in Scenedesmus is discussed.     

Mathematical Simulation of Electron Transport in the Primary Photosynthetic Processes

Biochemistry (Moscow) - Summarized results of investigation of regulation of electron transport and associated processes in the photosynthetic membrane using methods of mathematical and computer...     

Hydrogen Peroxide Inhibits Photosynthetic Electron Transport in Cells of Cyanobacteria

The effect of H₂O₂ on photosynthetic O₂ evolution and photosynthetic electron transfer in cells of cyanobacteria Anabaena variabilis and Anacystis nidulans was studied. The following experiments were performed: 1) directly testing the effect of exogenous H₂O₂; 2) testing the effect of intracellular H₂O₂ generated with the use of methyl viologen (MV); 3) testing the effect of inhibiting intracellular H₂O₂ decomposition by salicylic acid (SA) and 3-amino-1,2,4-triazole (AT). H₂O₂ inhibited photosynthetic O₂ evolution and light-induced reduction of p-benzoquinone (BQ) + ferricyanide (FeCy) in the Hill reaction. The I₅₀ value for H₂O₂ was 0.75 mM. Photosynthetic electron transfer in the cells treated with H₂O₂ was not maintained by H₂O₂, NH₂OH, 1,5-diphenylcarbazide, tetraphenylboron, or butylated hydroxytoluene added as artificial electron donors for Photosystem (PS) II. The H₂O CO₂, H₂O MV (involving PSII and PSI) and H₂O BQ + FeCy (chiefly dependent on PSII) electron transfer reactions were inhibited upon incubation of the cells with MV, SA, or AT. The N,N,N",N"-tetramethyl-p-phenylenediamine MV (chiefly dependent on PSI) electron transfer was inhibited by SA and AT but was resistant to MV. The results show that H₂O₂ inhibits photosynthetic electron transfer. It is unlikely that H₂O₂ could be a physiological electron donor in oxygenic photosynthesis.     

CHLOROPLAST STRUCTURE AND FUNCTION IN ac-20, A MUTANT STRAIN OF CHLAMYDOMONAS REINHARDI : II. Photosynthetic Electron Transport

Photosynthetic electron transport is markedly affected in mixotrophic cells of ac-20 because they lack the capacity to form the wild-type level of cytochrome 559, as well as Q, the quencher of fluorescence of photochemical system II. The other components of the electron-transport chain, as well as reactions dependent upon photochemical system I, are unaffected in the mutant strain. These observations are discussed in terms of the previously reported effects of the ac-20 mutation on CO₂ fixation and ribulose-1,5-diphosphate carboxylase activity.     

Effect of Light on the Rate of Glycolysis in Scenedesmus obliquus

The production of d-lactic acid, a major fermentation product of Scenedesmus obliquus strain D₃, is inhibited by light. This inhibition has been observed in the wild type as well as in Bishop's mutant 11. However, in mutant 8, which lacks photophosphorylation, light stimulated the formation of d-lactic acid.