Mitochondrial respiration of the photosynthesizing cell

Current notions on respiration of photosynthesizing cells are reviewed. Over the past three decades, the modern methods based on isotope techniques and reverse and molecular genetics provided convincing evidence that mitochondrial respiration is functional in the light and contributes to the creation of optimal conditions for photosynthesis and for protection of cells from photodegradation. Novel data are presented on the substrates that are used for respiration in the light. Individual respiration steps are considered in the context of their possible role in photosynthesizing cells. The mechanisms and carriers mediating the export of reducing equivalents from chloroplasts for their subsequent oxidation in the mitochondrial electron-transport chain are discussed. The regulation of nonphosphorylating (unrelated to energy generation) electron transport pathways mediated by alternative oxidase and alternative type II NADPH-dehydrogenases, as well as the role of uncoupling proteins in plant mitochondria, are analyzed. These components were shown to play a significant role in NAD(P)H oxidation for maintaining the redox balance in mitochondria and whole green cells. A generalized scheme of biochemical interactions between organelles—chloroplasts, mitochondria, and peroxisomes—is presented. The directions for future research are outlined.     

Comparison of oxygen consumption rates in minimally transformed BALB/3T3 and virus-transformed 3T3B-SV40 cells

In the recent years, bioenergetics of tumor cells and particularly cell respiration have been attracting great attention because of the involvement of mitochondria in apoptosis and growing evidence of the possibility to diagnose and treat cancer by affecting the system of oxidative phosphorylation in mitochondria. In the present work, a comparative study of oxygen consumption in 3T3B-SV40 cells transformed with oncovirus SV40 and parental BALB/3T3 cells was conducted. Such fractions of oxygen consumption as “phosphorylating” respiration coupled to ATP synthesis, “free” respiration not coupled to ATP synthesis, and “reserve” or hidden respiration observed in the presence of protonophore were determined. Maximal respiration was shown to be only slightly decreased in 3T3B-SV40 cells as compared to BALB/3T3. However, in the case of certain fractions of cellular respiration, the changes were significant. “Phosphorylating” respiration was found to be reduced to 54% and “reserve” respiration, on the contrary, increased up to 160% in virus-transformed 3T3B-SV40 cells. The low rate of “phosphorylating” respiration and high “reserve” respiration indicate that under normal incubation conditions the larger part of mitochondrial respiratory chains of the virus-transformed cells is in the resting state (i.e. there is no electron transfer to oxygen). The high “reserve” respiration is suggested to play an important role in preventing apoptosis of 3T3B-SV40 cells.     

The rate of photorespiration as measured by means of oxygen uptake and its respiratory quotient

Oxygen recycling inside photosynthesizing leaves was found to amount to less than 0.3% of the oxygen consumed by photorespiration under natural conditions, provided the influence of buildup of oxygen released by photosynthesis into the external air was taken into consideration. When this is related to the amounts of photorespired CO₂, which had been previously found to be reabsorbed by photosynthesis, it appears that previous respiratory quotients reported for photorespiration were underestimated. For the same reason the photosynthetic quotient was overestimated. Actually, quotients of photorespiration and of photosynthesis approach the more normal range of respiratory quotients int the dark. The oxygen recycling was calculated according to an electrical analogue to oxygen flow.     

PHENOTYPIC VARIATION WITHIN “INDIVIDUAL” LICHEN THALLI

“Individual” large diam lichen thalli of Umbilicaria vellea (L.) Ach. and U. mammulata (Ach.) Tuck. are not uniform in physiological activity or enzymatic phenotype. Horizontal variation in net photosynthetic rates and dark respiration rates was found, but this was independent of the mass of the photosynthesizing or respiring tissues. In addition, sublethal temperature pretreatments reduced the net photosynthetic and respiratory rates of some areas of the thallus, but increased others. Assays of isoenzyme phenotypes showed that all large diam thalli are extensively polymorphic while young, small diam thalli are mainly monomorphic. Taken together, the results say that the concept of the “individual” may be inappropriate for these organisms.     

Metabolic scaling theory in plant biology and the three oxygen paradoxa of aerobic life

Alfred Russell Wallace was a field naturalist with a strong interest in general physiology. In this vein, he wrote that oxygen (O₂), produced by green plants, is “the food of protoplasm, without which it cannot continue to live”. Here we summarize current models relating body size to respiration rates (in the context of the metabolic scaling theory) and show that oxygen-uptake activities, measured at 21 vol.% O₂, correlate closely with growth patterns at the level of specific organs within the same plant. Thus, whole plant respiration can change ontogenetically, corresponding to alterations in the volume fractions of different tissues. Then, we describe the evolution of cyanobacterial photosynthesis during the Paleoarchean, which changed the world forever. By slowly converting what was once a reducing atmosphere to an oxidizing one, microbes capable of O₂-producing photosynthesis modified the chemical nature and distribution of the element iron (Fe), slowly drove some of the most ancient prokaryotes to extinction, created the ozone (O₃) layer that subsequently shielded the first terrestrial plants and animals from harmful UV radiation, but also made it possible for Earth’s forest to burn, sometimes with catastrophic consequences. Yet another paradox is that the most abundant protein (i.e., the enzyme Rubisco, Ribulose-1,5-biphosphate carboxylase/oxygenase) has a greater affinity for oxygen than for carbon dioxide (CO₂), even though its function is to bind with the latter rather than the former. We evaluate this second “oxygen paradox” within the context of photorespiratory carbon loss and crop yield reduction in C3 vs. C4 plants (rye vs. maize). Finally, we analyze the occurrence of reactive oxygen species (ROS) as destructive by-products of cellular metabolism, and discuss the three “O₂-paradoxa” with reference to A. R. Wallace’s speculations on “design in nature”.     

Light-enhanced dark respiration in leaves and mesophyll protoplasts of pea in relation to photorespiration,respiration and some metabolites content

The respiration rate of leaves and mesophyll protoplasts of pea (Pisum sativum L.), from plants which were previously kept in darkness for 24 h was doubled following a period of photosynthesis at ambient level of O₂ (21 %), whereas the low level of O₂ (1 % and 4 % for leaves and protoplasts, respectively) reduced this light-enhanced dark respiration (LEDR) to the rate as noted before the illumination. Similarly to respiration rate, the oxygen at used concentrations had no effect on the ATP/ADP ratio in the dark-treated leaves. However, the ATP/ADP ratio in leaves photosynthesizing at 21 % O₂ was higher (up to 40 %, dependence on CO₂ concentration in the range 40–1600 1 dm⁻³) than in those photosynthesizing at 1 % O₂ or darkened at air (21 % O₂). Also, at 1 % O₂ the accumulation of malate was suppressed (by about 40 %), to a value noted for leaves darkened at 21 % O₂. The dark-treatment of leaves reduced the ability of isolated mitochondria to oxidize glycine (by about twofold) and succinate, but not malate. Mitochondria from both the light- and dark-treated leaves did not differ in qualitative composition of free amino acids, however, there were significant quantitative differences especially with respect to aspartate, alanine, glutamate and major intermediates of the photorespiratory pathway (glycine, serine). Our results suggest that accumulation of photorespiratory and respiratory metabolites in pea leaves during photosynthesis at 1 % O₂ is reduced, hence the suppression of postillumination respiration rate.     

Changes in the functional organization of the respiratory system in residents of Western Siberia in the winter season

The respiratory parameters of healthy young men from Western Siberia have been examined. The correlations between the parameters have been analyzed, and the functional structure of the respiratory system in summer and winter has been identified. It has been found that different regulatory programs operate depending on the temperature of the inhaled air. The study has shown that the changes in the oxygen demand of the body in summer are mediated by the changes in the volume of pulmonary ventilation (“ventilation” or “summer program”). In winter, when maintaining the level of energy processes in the body is in conflict with maintaining thermal homeostasis in the respiratory regions of the lungs, pulmonary ventilation becomes limited, and the number of functioning lung units is reduced. At the same time, in compensation, the lung diffusion capacity increases (“diffusion” or “winter program”). This means that the functioning of the apparatus of external respiration is optimized in winter.     

Multicellularity arose several times in the evolution of eukaryotes (Response to DOI 10.1002/bies.201100187)

The cellular slime mold Dictyostelium has cell‐cell connections similar in structure, function, and underlying molecular mechanisms to animal epithelial cells. These similarities form the basis for the proposal that multicellularity is ancestral to the clade containing animals, fungi, and Amoebozoa (including Dictyostelium): Amorphea (formerly “unikonts”). This hypothesis is intriguing and if true could precipitate a paradigm shift. However, phylogenetic analyses of two key genes reveal patterns inconsistent with a single origin of multicellularity. A single origin in Amorphea would also require loss of multicellularity in each of the many unicellular lineages within this clade. Further, there are numerous other origins of multicellularity within eukaryotes, including three within Amorphea, that are not characterized by these structural and mechanistic similarities. Instead, convergent evolution resulting from similar selective pressures for forming multicellular structures with motile and differentiated cells is the most likely explanation for the observed similarities between animal and dictyostelid cell‐cell connections.     

Effect of oxygen concentration on photosynthesis and respiration in two hypersaline microbial mats

The effects of oxygen concentration on photosynthesis and respiration in two hypersaline cyanobacterial mats were investigated. Experiments were carried out on mats from Eilat, Israel, with moderate photosynthetic activity, and mats from Mallorca, Spain, with high photosynthetic activity. The oxygen concentration in the overlying water above the mats was increased stepwise from 0% to 100% O2. Subsequent changes in oxygen concentration, gross photosynthetic rates, and pH values inside the mats were measured with microelectrodes. According to published reports on the regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the key enzyme in the CO2-fixation pathway of phototrophs, we expected photosynthetic activity to decrease with increasing oxygen concentration. Gross photosynthetic and total respiration rates in both mats were highest when the O2 concentration was at 0% in the overlying water. Net oxygen production rates under these conditions were the same as under air saturation (21% O2), while gross photosynthetic and respiration rates were lowest at air saturation. In both mats, gross photosynthetic and respiration rates increased upon gradually increasing the oxygen concentration in the overlying water from 21% to 100%. These results contradict the expectation that photosynthesis decreases with increasing oxygen concentration. Increased photosynthetic rates at oxygen concentrations above 21% were probably caused by enhanced oxidation of organic matter and concomitant CO2 production due to the increased oxygen availability. The cause of the high respiration rates at 0% O2 in the overlying water was presumably the enhanced excretion of photosynthetic products during increased photosynthesis. We conclude that the effect of the O2/CO2 concentration ratio on the activity of Rubisco as demonstrated in vitro on enzyme extracts cannot be extrapolated to the situation in intact microbial mats, because the close coupling of the activity of primary producers and heterotrophic bacteria plays a major role in this ecosystem.     

Crown group Oxyphotobacteria postdate the rise of oxygen

The rise of oxygen ca. 2.3 billion years ago (Ga) is the most distinct environmental transition in Earth history. This event was enabled by the evolution of oxygenic photosynthesis in the ancestors of Cyanobacteria. However, long‐standing questions concern the evolutionary timing of this metabolism, with conflicting answers spanning more than one billion years. Recently, knowledge of the Cyanobacteria phylum has expanded with the discovery of non‐photosynthetic members, including a closely related sister group termed Melainabacteria, with the known oxygenic phototrophs restricted to a clade recently designated Oxyphotobacteria. By integrating genomic data from the Melainabacteria, cross‐calibrated Bayesian relaxed molecular clock analyses show that crown group Oxyphotobacteria evolved ca. 2.0 billion years ago (Ga), well after the rise of atmospheric dioxygen. We further estimate the divergence between Oxyphotobacteria and Melainabacteria ca. 2.5–2.6 Ga, which—if oxygenic photosynthesis is an evolutionary synapomorphy of the Oxyphotobacteria—marks an upper limit for the origin of oxygenic photosynthesis. Together, these results are consistent with the hypothesis that oxygenic photosynthesis evolved relatively close in time to the rise of oxygen.     

Components of CO<Subscript>2</Subscript> exchange in leaves of C<Subscript>3</Subscript> species with different ability of starch accumulation

Using a radiogasometric method the rates of photorespiratory and respiratory decarboxylations of primary and stored photosynthates in the leaves of two groups of C₃ species, differing in the ability of starch accumulation, were determined. One group included starch-accumulating (SA) species with rates of starch synthesis on the average 38 % the rate of photosynthesis [Solanum tuberosum L., Arabidopsis thaliana (L.) Heynh, Helianthus annuus L., and Plantago lanceolata L.]. The second group represented starch-deficient (SD) species with rates of starch synthesis less than 8 % the rate of photosynthesis (Secale cereale L., Triticum aestivum L., Hordeum vulgare L., and Poa trivialis L.). In SA species the rate of respiration in the dark was significantly higher than in SD species. No differences were found in the rates of photosynthesis, photorespiration, and respiration under irradiation. Thus, the degree of inhibition of respiration by irradiation was in SA species higher than in SD species. It is concluded that starch does not provide substrates for respiratory and photorespiratory decarboxylations in irradiated photosynthesizing leaves.     

Veränderungen des Stoffwechsels der Ascorbin- und Glycolsäure inNicotiana tabacum L.—Pflanzen bei Infektion mit dem Virus der Kartoffelstrichelkrankheit

Changes in the metabolic pathway linked up directly with respiratory pathways were followed inN. tabacum L. cv. ‘Samsun’ plants inoculated with potato Y virus. The consumption of oxygen in infected tissues before the occurrence of visible symptoms (up to the sixth day) was lower than in control plants, then it started to rise and the tenth day after inoculation it reached 116% of the control. It was established that two main respiratory chains were involved, from which the glutathion-ascorbate system was responsible for the total respiration rate during the entire period of investigation. The other respiratory system,i.e. NADPH—glyoxalate—glycolate, is not related directly to the respiration rate, but, it is probably linked up with the changing content of NADPH. The results obtained with catalase and peroxidase show that these enzymes are not directly linked up with changing glycollate oxidase activity but that they cooperate. While at the beginning their activities are mutually compensated, at the later period of enhanced glycollate oxidase activity and increase in the activities of both these enzymes occurs which then remove the originating excessive toxic H₂O₂.     

Catalase characterization and implication in bleaching of a symbiotic sea anemone

Symbiotic cnidarians are marine invertebrates harboring photosynthesizing microalgae (named zooxanthellae), which produce great amounts of oxygen and free radicals upon illumination. Studying antioxidative balance is then crucial to understanding how symbiotic cnidarians cope with ROS production. In particular, it is suspected that oxidative stress triggers cnidarian bleaching, i.e., the expulsion of zooxanthellae from the animal host, responsible for symbiotic cnidarian mass mortality worldwide. This study therefore investigates catalase antioxidant enzymes and their role in bleaching of the temperate symbiotic sea anemone Anemonia viridis. Using specific separation of animal tissues (ectoderm and endoderm) from the symbionts (zooxanthellae), spectrophotometric assays and native PAGE revealed both tissue-specific and activity pattern distribution of two catalase electrophoretypes, E1 and E2. E1, expressed in all three tissues, presents high sensitivity to the catalase inhibitor aminotriazole (ATZ) and elevated temperatures. The ectodermal E1 form is responsible for 67% of total catalase activity. The E2 form, expressed only within zooxanthellae and their host endodermal cells, displays low sensitivity to ATZ and relative thermostability. We further cloned an ectodermal catalase, which shares 68% identity with mammalian monofunctional catalases. Last, 6 days of exposure of whole sea anemones to ATZ (0.5 mM) led to effective catalase inhibition and initiated symbiont expulsion. This demonstrates the crucial role of this enzyme in cnidarian bleaching, a phenomenon responsible for worldwide climate-change-induced mass mortalities, with catastrophic consequences for marine biodiversity.     

轻度水分胁迫的小麦幼苗中与呼吸有关的几种酶活性变化

轻度水分胁迫使小麦幼苗叶片呼吸升高时,叶中琥珀酸去氢酶和细胞色素氧化酶活性均明显升高;而同样胁迫使根呼吸下降时,根中这两种酶活性均明显下降。叶和根中ATP酶分解活性在胁迫下都明显升高。轻度水分胁迫使叶片过氧化氢酶活性升高。叶中有明显的乙醇酸氧化酶活性,抗旱品种的酶活性较高,胁迫使此酶活性降低。     

ECOPHYSIOLOGICAL PERFORMANCE OF THE AEROTERRESTRIAL GREEN ALGA KLEBSORMIDIUM CRENULATUM (CHAROPHYCEAE,STREPTOPHYTA) ISOLATED FROM AN ALPINE SOIL CRUST WITH AN EMPHASIS ON DESICCATION STRESS1

Aeroterrestrial filamentous green algae of the genus Klebsormidium (Klebsormidiales, Streptophyta) are typical components of biological soil crusts, which occur worldwide in arid and semiarid habitats including alpine regions. In the present study, Klebsormidium crenulatum (Kütz.) Lokhorst was isolated from an alpine soil crust above the timberline of the Austrian Alps. Growth responses, photosynthetic performance, and desiccation tolerance were measured under controlled laboratory conditions. K. crenulatum exhibited optimal growth and the highest photosynthetic efficiency under relatively low photon fluence densities (30 and 21.9 μmol photons · m^?2 · s^?1, respectively), indicating low‐light requirements. It grew in a narrow range of salinities between 1.2 and 15 practical salinity units (psu), pointing to a pronounced stenohaline response pattern. Increasing temperatures from 5°C to 40°C led to different effects on photosynthetic oxygen evolution and respiratory oxygen consumption in K. crenulatum. While at low temperatures (5°C–10°C) photosynthesis was relatively high, respiration was not detectable or was at a very low level. Conversely, at the highest temperature of 40°C, photosynthesis was inhibited, and respiration unaffected, indicating strong differences in temperature sensitivity between both physiological processes. K. crenulatum was capable of photosynthesizing efficiently for up to 2.5 h under desiccation, followed by a decrease to 15% of the initial value after 3 h. Complete recovery took place within 2 h after rehydration. All ecophysiological data explain the widespread abundance of K. crenulatum in soil crusts of the alpine regions of the European Alps.     

Beneficial interaction between photosynthesis and respiration in mesophyll protoplasts of pea during short light-dark cycles

The respiratory uptake or photosynthetic evolution of oxygen by mesophyll protoplasts of pea ( Pisum sativum L. cv. Arkel) were monitored during successive short. (3–5 min) cycles of darkness and illumination. The rate of respiration was nearly doubled after 3–4 short periods of illumination while there was a 15–20% enhancement in photosynthesis with cycles of illumination and darkness preceding illumination. Such interaction between photosynthesis and respiration was statistically significant when bicarbonate was present in the reaction medium. The inhibitors of photosynthesis [3(3,4–dichlorophenyl)-l,l-dimethylurea (DCMU), glyceraldehyde] decreased respiration after periods of illumination, whereas inhibitors of respiratory electron transport (Rotenone, antimycin A, NaN₃) suppressed photosynthesis, as well. We suggest that a rapid beneficial interaction exists between photosynthesis and respiration in protoplasts, even during short cycles of light and darkness.     

The effect of venturicidin on light and oxygen-dependent electron transport,proton translocation,membrane potential development and ATP synthesis in intact cells of Rhodopseudomonas capsulata

Venturicidin behaves as an orthodox energy transfer inhibitor in intact cells of Rhodopseudomonas capsulata as judged by the following criteria. 1. It led to inhibition of respiration. Inhibition was relieved by low concentrations of uncoupling agent. 2. It enhanced light-induced and oxygen dependent H⁺ efflux. 3. It stimulated light-induced and oxygen dependent carotenoid band shifts. The rate of decay of the band shifts after short flash excitation was decreased in the presence of venturicidin. 4. It stimulated light-induced and oxygen dependent butyltriphenylphosphonium uptake. 5. It inhibited the rise in cellular ATP concentration accompanying either photosynthesis or respiration.     

Profound afternoon depression of ecosystem production and nighttime decline of respiration in a macrophyte-rich,shallow lake

Small, shallow lakes with dense growth of submerged macrophytes are extremely abundant worldwide, but have remained grossly understudied although open water oxygen measurements should be suitable to determine diel fluctuations and test drivers of ecosystem metabolism during the day. We measured the temporal and spatial variability of environmental conditions as well as net ecosystem production (NEP) and respiration (R) in a small, shallow Swedish lake with dense charophyte stands by collecting data from oxygen-, pH-, temperature- and light-sensors across horizontal and vertical gradients during different periods between April and June in 3 years. We found reproducible diel oxygen patterns and daily metabolic rates. The charophyte canopy accounted for almost all primary production and respiration of the ecosystem. Two novel discoveries—profound afternoon depression of production and nighttime decline of respiration—occurred on virtually every day. Extensive increase of oxygen-, temperature- and pH-levels and depletion of dissolved inorganic carbon (DIC) and CO₂ concentrations could account for maximum NEP-rates before noon and afternoon depression with low NEP-rates. Ecosystem respiration declined during the night to 24–70% of rates at sunset, probably because of depletion of respiratory substrates. Afternoon depression of photosynthesis should be widespread in numerous habitats with dense growth of macrophytes, periphyton, or phytoplankton implying that daily photosynthesis and growth are restricted and species with efficient DIC use may have an advantage.     

The Cold Tolerance of Indica Type Rice in Relation to Its Parents

To understand the inheritance of cold tolerant characteristics of rice (Oryza sativa L. ), the authors investigated the cold tolerance of hybrid progenies in relation to their parental lines which were "Qinghua 6—a cold sensitive cultivar and "Guishan’aixuan 3”— a cold tolerant cultivar. The hybrid progenies were the cross and the reciprocal cross of these two cultivars. Results showed that after 1℃ dark or light (250 μmol · m-2 · s-1) treatment, the survival rate of seedlings was higher in "Guishan’aixuan 3” than that in "Qinghua 6”. That of the hybrid "Qinghua 6” ( ♂ )× "Guishan’aixuan 3” (♀) was higher than that . in hybrid "Guishan’aixuan 3” ( ♂ ) × "Qinghua 6” (♀). Detached flag leaves at heading stage when treated in light and chilling condition for 12, 24, 36 h, there was also less decrease of photosynthesis in "Guishan’aixuan 3” and "Qinghua 6” ( ♂ ) × "Guishan’aixuan 3” (♀) than in “Qinghua 6” and “Guishan’aixuan 3” ( ♂ ) × “Qinghua 6” ( ♀ ). After being treated for 12 h, respiratory rate rose in the former but not the latter. Changes of fluorescence parameters, rise of Fo and decline of Fm and Fv, were caused by 1 ℃ and light (250 μmol·m-2·s-1) treatment. The decrease of Fv/Fo and Fv/Fm were more apparent in the former than in the latter after light and chilling treatment for 24 h, with a much faster recovery under normal temperature in the former than in the latter. Effect of the natural low temperature (cold, dew and wind) on the chlorophyll fluorescence of rice was similar to that of artificial treatment. It was suggested that cold tolerance of rice progenies seemed to be from maternal inheritance.     

Inhibition of respiration by sodium fluoride and monoiodoacetic acid in wheat leaves at different oxygen tensions and in the presence of methylene blue and L-cysteine

Six-day leaves of wheat (Triticum vulgare) were used to test the effect of oxygen, nitrogen and different mixtures of these gases, methylene blue and L-cysteine on the inhibition of respiration by sodium fluoride and monoiodoacetic acid using the Warburg manometric method. In an atmosphere of 100% oxygen total respiration was increased by an average of 45%, 10% oxygen led to a decrease in respiration by 20%. However, no simultaneous changes were found in the degree of inhibition by these enzyme poisons. On decreasing the oxygen content to 5% inhibition of respiration was almost doubled while at the same time respiration fell by an average of 23%. In nitrogen, the respiration rate varied around 10% and inhibition was almost 100%. The results showed that the action of enzyme poisons is only influenced by different oxygen content at low oxygen tension and is evidently associated with the dominance of a given energy system (glycolysis). On using methylene blue and L-cystein in concentrations of 10-3M a decrease in the relative inhibition of respiration by enzyme poisons was also found. The total respiration rate, however, was only raised by 6–8%. Since the change in the degree of inhibition on using these substances is caused by an increase in the participation of respiratory processes resistant to glycolytic inhibitors it can, in this case be assumed that there is an increase in the activity of the pentose cycle in such tissue.