Participation of Mitochondrial Metabolism in Photorespiration : Reconstituted System of Peroxisomes and Mitochondria from Spinach Leaves

In this study the interplay of mitochondria and peroxisomes in photorespiration was simulated in a reconstituted system of isolated mitochondria and peroxisomes from spinach (Spinacia oleracea L.) leaves. The mitochondria oxidizing glycine produced serine, which was reduced in the peroxisomes to glycerate. The required reducing equivalents were provided by the mitochondria via the malate-oxaloacetate (OAA) shuttle, in which OAA was reduced in the mitochondrial matrix by NADH generated during glycine oxidation. The rate of peroxisomal glycerate formation, as compared with peroxisomal protein, resembled the corresponding rate required during leaf photosynthesis under ambient conditions. When the reconstituted system produced glycerate at this rate, the malate-to-OAA ratio was in equilibrium with a ratio of NADH/NAD of 8.8 × 10⁻³. This low ratio is in the same range as the ratio of NADH/NAD in the cytosol of mesophyll cells of intact illuminated spinach leaves, as we had estimated earlier. This result demonstrates that in the photorespiratory cycle a transfer of redox equivalents from the mitochondria to peroxisomes, as postulated from separate experiments with isolated mitochondria and peroxisomes, can indeed operate under conditions of the very low reductive state of the NADH/NAD system prevailing in the cytosol of mesophyll cells in a leaf during photosynthesis.     

Distinction between light-mediated and light-independent variations in phytoplankton production rates

An array of factors simultaneously controls phytoplankton photosynthesis and hence the primary production process. Because their relative importance shifts both with depth and with season, the significance of individual factors cannot be resolved by in situ incubations, even if all relevant environmental and biotic variables are measured.Here a procedure is described by which in addition to in situ measurements, photosynthesis is simultaneously assessed in identical subsamples under constant temperature (10 °C) and light (0.66 mol m^–2 h^–1 PAR conditions, in vitro). By calculating photosynthesis per unit of chlorophyll, effects of shifting biomass on photosynthesis can be eliminated but seasonal variations of light-saturated photosynthesis generated by temperature, and vertical changes of light-requirements (e.g. by light-shade adaptation) remain obscure. Quotients of in situ photosynthetic rates divided by in vitro rates allow the quantification of light-mediated changes. Provided that photosynthesis measured in vitro is light-saturated, quotients in situ: in vitro rates should never exceed unity. They are a measure for the degree of light-limitation. In vitro rates normalized to chlorophyll give information on temporal changes caused by variations in photosynthetic capacity. In Lake Constance, mean cell size appears to control light-saturated assimilation numbers.     

Leaf‐Mosaic‐Inspired Vine‐Like Graphitic Carbon Nitride Showing High Light Absorption and Efficient Photocatalytic Hydrogen Evolution

Green plants use solar energy efficiently in nature. Simulating the exquisite structure of a natural photosynthesis system may open a new approach for the construction of desirable photocatalysts with high light harvesting efficiency and performance. Herein, inspired by the excellent light utilization of “leaf mosaic” in plants, a novel vine‐like g‐C₃N₄ (V‐CN) is synthesized for the first time by copolymerizing urea with dicyandiamide‐formaldehyde (DF) resin. The as‐prepared V‐CN exhibits ultrahigh photocatalytic hydrogen production of 13.6 mmol g^?1 h^?1 under visible light and an apparent quantum yield of 12.7% at 420 nm, which is ≈38 times higher than that of traditional g‐C₃N₄, representing one of the highest‐activity g‐C₃N₄‐based photocatalysts. This super photocatalytic performance is derived from the unique leaf mosaic structure of V‐CN, which effectively improves its light utilization and affords a larger specific surface area. In addition, the introduction of DF resin further optimizes the energy band of V‐CN, extends its light absorption, and improves its crystallinity and interfacial charge transport, resulting in high performance. It is an easy and green strategy for the preparation of broad‐spectrum, high‐performance g‐C₃N₄, which presents significant advancement for the design of other nanophotocatalysts by simulating the fine structure of natural photosynthesis.     

Photosynthesis, Photorespiration and Respiration of Detached Spruce Twigs as Influenced by Oxygen Concentration and Light Intensity

The effects of oxygen concentration and light intensity on the rates of apparent photosynthesis, true photosynthesis, photorespiration and dark respiration of detached spruce twigs were determined by means of an infra-red carbon dioxide analyzer (IRCA). A closed circuit system IRCA was filled with either 1 per cent of oxygen in nitrogen, air (21 % O₂) or pure oxygen (100 % O₂). Two light intensities 30 × 10³ erg · cm ^?2· s^?1 and 120 × 10³ erg · cm^?2· s^?1 were applied. It has been found that the inhibitory effect of high concentration of oxygen on the apparent photosynthesis was mainly a result of a stimulation of the rate of CO₂ production in light (photorespiration). In the atmosphere of 100 % O₂, photorespiration accounts for 66–80 per cent of total CO₂ uptake (true photosynthesis). Owing to a strong acceleration of photorespiration by high oxygen concentrations, the rate of true photosynthesis calculated as the sum of apparent photosynthesis and photorespiration was by several times less inhibited by oxygen than the rate of apparent photosynthesis. The rates of dark respiration were essentially unaffected by the oxygen concentrations used in the experiments. An increase in the intensity of light from 30 × 10³ erg · cm^?3· s^?1 to 120 · 10³ erg · cm^?2· s^?1 enhanced the rate of photorespiration in the atmospheres of 21 and 100 % oxygen but not in 1 % O₂. The rate of apparent photosynthesis, however, was little affected by light intensity in an atmosphere of 1 % oxygen.     

Mitochondria contribute to increased photosynthetic capacity of leaves of winter rye (Secale cereale L.) following cold-hardening

Cold-hardening of winter rye (Secale cereale L. cv. Musketeer) increased dark respiration from ?2.2 to ?3.9 μmol O₂ m^?2s^?1 and doubled light-and CO₂-saturated photosynthesis at 20°C from 18.1 to 37.0μmol O₂ m^?2 s^?1 We added oligomycin at a concentration that specifically inhibits oxidative phosphorylation to see whether the observed increase in dark respiration reflected an increase in respiration in the light, and whether this contributed to the enhanced photosynthesis of cold-hardened leaves. Oligomycin inhibited light- and CO₂-saturated rates of photosynthesis in non-hardened and cold-hardened leaves by 14 and 25%, respectively, and decreased photochemical quenching of chlorophyll a fluorescence to a greater degree in cold-hardened than in non-hardened leaves. These data indicate an increase both in the rate of respiration in the light, and in the importance of respiration to photosynthesis following cold-hardening. Analysis of metabolite pools indicated that oligomycin inhibited photosynthesis by limiting regeneration of ribulose-1,5-bisphosphate. This limitation was particularly severe in cold-hardened leaves, and the resulting low 3-phospho-glycerate pools led to a feed-forward inhibition of sucrose-phosphate synthase activity. Thus, it does not appear that oxidative phosphorylation supports the increase in photo-synthetic O₂ evolution following cold-hardening by increasing the availability of cytosolic ATP. The data instead support the hypothesis that the mitochondria function in the light by using the reducing equivalents generated by non-cyclic photosynthetic electron transport.     

Primary Productivity of Phytoplankton,Environmental Covariates and Fish Growth in West Bengal Ponds under a Polyculture System

Measurements of primary production by phytoplankton in four fish ponds over a period of one year revealed a distinct spatial variation in rate. The ponds were used for polyculture, monoculture, or a traditional system of fish culture. The rates of gross photosynthesis were highly correlated with temperature, free CO₂, oxygen, inorganic nitrogen (NH₄⁺, NO₂⁻, and NO₃⁻) and PO₄-P. It was shown by step-wise multiple regression analysis that in the polyculture pond with a predominantly autotrophic metabolism, gross photosynthesis was the most significant factor correlated with fish growth. In another polyculture system in which the metabolism was mainly hetero-trophic, the influence of the photosynthesis rate was slight.     

Experiments in whole leaf photosynthesis

The process of photosynthesis is of such importance that most courses of biochemistry and physiology include some reference to it. This paper describes a simple experimental system, which uses radioactive carbon dioxide to study whole leaf photosynthesis under a variety of conditions.

An apparatus of very simple construction, in which the experiments can be safely carried out, is described. The experiments basically consist of the exposure of leaves to ¹⁴CO₂ and the subsequent determination of radioactivity distribution and intensity by counting of leaf discs in a Geiger-Muller system and/or by autoradiography.

Experiments to demonstrate photosynthesis in the presence and absence of light, in variegated leaves of different types and to illustrate the importance of stomata are described.     

Lidocaine and ATPase inhibitor interaction with the chloroplast envelope

Photosynthetic capacity of isolated intact chloroplasts is known to be sensitive to K⁺ fluxes across the chloroplast envelope. However, little is known about the system of chloroplast envelope proteins that regulate this K⁺ movement. The research described in this report focused on characterizing some of the components of this transport system by examining inhibitor effects on chloroplast metabolism. Digitoxin, an inhibitor of membrane-bound Na⁺/K⁺ ATPases, was found to reduce stromal K⁺ at a range of external K⁺ and inhibit photosynthesis. Scatchard plot analysis revealed a specific protein receptor site with a K_m for digitoxin binding of 13 nanomolar. Studies suggested that the receptor site was on the interior of the envelope. The effect of a class of amine anesthetics that are known to be K⁺ channel blockers on chloroplast metabolism was also studied. Under conditions that facilitate low stromal pH and concomitant photosynthetic inhibition, the anesthetic, lidocaine, was found to stimulate photosynthesis. This stimulation was associated with the maintenance of higher stromal K⁺. Comparison of the effects on photosynthesis of lidocaine analogs which varied in lipophilicity suggested a lipophilic pathway for anesthetic action. The results of experiments with lidocaine and digitoxin were consistent with the hypothesis that a K⁺ channel and a K⁺-pumping envelope ATPase contribute to overall K⁺ flux across the chloroplast envelope. Under appropriate assay conditions, photosynthetic capacity of isolated chloroplasts was shown to be much affected by the activity of these putative envelope proteins.     

Design of a Calorimeter for the Continuous Study of Heat Production during Anaerobic Microbial Growth

A conduction type calorimeter has been designed to chase microbial growth in batch system. The calorimeter is of a twin structure having thermopile plates as a temperature sensor, The heat evolution during the microbial growth at a required temperature can be observed as an output-voltage generated at thermopile terminals with a sensitivity of 58.5 mV K^?1

A stainless steel cell with a volume of 300 cm³ serves as a culture cell which is capable of being autoclaved prior to the initiation of calorimetric run, taking out from the calorimeter body.

Because of the twin structure, the apparatus works with sufficient stability in detecting small heat evolution for long duration. Its operation has been demonstrated with the growth of Sacch. cerevisiae grown on liquid synthetic medium under anaerobic condition.     

The Uptake of Free CO2 and HCO3- during Photosynthesis of Plankton Algae with Special Reference to the Coccolithophorid Coccolithus huxleyi

From a re-evaluation of experiments with the coccolithophorid Coccolithus hurleyi made by Paasche (1964), curves are presented showing the rate of photosynthesis as a function of the concentration of both free CO₂ and bicarbonate CO₂. It is shown that photosynthesis in a naked clone is due only to the uptake of free CO₂. The problem concerning the high concentration of free CO₂ necessary for photosynthesis in Coccolithus huxleyi is discussed. It is shown that it is not due to lack of the enzyme carbonic anhydrase. Hence it is probable that the formation of coccoliths is the mechanism by which, in Coccolithus, the utilization of HCO₃^- ions in photosynthesis becomes possible. The OH^- ions produced during photosynthesis (₊Ca²⁺ taken up together with the HCO₃^- ions) are thus neutralized. This situation is different from other aquatics utilizing HCO₃^- in photosynthesis. Here the OH^- ions are neutralized via an ion exchange of Ca²⁺ with H⁺ in the surrounding medium.     

Gas exchange of a desert shrub (Zygophyllum dumosum Boiss.) under different soil moisture regimes during summer drought

The effects of soil water potential on photosynthesis and transpiration of whole Zygophyllum dumosum Boiss. shrubs were examined with a field IRGA system during a rainless summer. Daily photosynthesis and transpiration activities were not notably different on a unit phyllode area basis among shrubs at naturally differing soil water potentials. Irrigation of shrubs caused phyllodes to increase significantly in water content and new leaflets to appear. Leaflets had three times as many stomata per unit area (23000 stomata cm^-2) as phyllodes (7100 stomata cm^-2) but photosynthesis and transpiration rates were not measurably different between irrigated and non-irrigated shrubs on a unit area basis. This finding suggests that sufficient soil moisture will lead to increased carbon uptake of the entire shrub simply because the total area of photosynthesizing tissue increases. Gas exchange rates appear to be controlled solely by atmospheric conditions under the stresses of summer.     

Singlet oxygen inhibits ATPase and proton translocation activity of the thylakoid ATP synthase CF1CFo

Singlet oxygen (¹O₂) produced in plants during photosynthesis has a strong damaging effect not only on both photosystems but also on the whole photosynthetic machinery. This is also applicable for the adenosine triphosphate (ATP) synthase. Here we describe the impact of ¹O₂ generated by the photosensitizer Rose Bengal on the ATP hydrolysis and ATP-driven proton translocation activity of CF1CFo. Both activities were reduced dramatically within 1 min of exposure. Interestingly, it is shown that oxidized thylakoid ATP synthase is more susceptible to ¹O₂ than CF1CFo in its reduced state, a new insight on the mechanism of ¹O₂ interaction with the γ subunit.     

Seasonal Course of Photosynthesis, Respiration, and Distribution of C in Young Pinus resinosa Trees as Related to Wood Formation

Rates of net photosynthesis and dark respiration, and distribution of ¹⁴C were determined for new (current season's) and old (previous season's) needles at 10 times during the seasonal development of young Pinus resinosa Ait. trees. The seasonal changes in these factors associated with the development of the new shoot were related to known seasonal patterns of wood formation.

Net photosynthesis per gram of needle dry weight (photosynthetic efficiency) was maximum in the old needles at the time of first new needle elongation; at the same time translocation of ¹⁴C from old to new needles was greatest. Photosynthetic efficiency of new needles was maximum at the end of the period of rapid new needle elongation, when the new needles also began exporting much greater quantities of ¹⁴C to other plant parts. In particular, the amount translocated from the new needles to the stem was greatly increased. At this time thick-walled xylem cells were first observed in the stem.

These results, together with those of previous studies, indicate that the production of thick-walled xylem tracheids normally associated with latewood is physiologically correlated with maturation of the current season's needles. Because there is a lesser demand for photosynthate in the new shoot and a high rate of photosynthesis in the whole plant at the time of new needle maturity, a sharply increased amount of photosynthate becomes available for wall synthesis by cambial derivatives.

     

Line-shape analysis of polarized electron paramagnetic resonance spectra of the primary reactants of bacterial photosynthesis

A simple analytical method is described for calculating transient EPR spectra of the primary reactants of photosynthesis. Polarization is generated in the primary radical pair P⁺I⁻ and passed to the acceptor X⁻ by electron transfer. Satisfactory agreement between theoretical and experimental spectra is obtained by including a dipolar interaction between P⁺ and I⁻ and an anisotropic g tensor for X⁻.     

Influence of carbon supply on the circadian rhythmicity of photosynthesis and its stimulation by blue light in Ectocarpus siliculosus: clues to the mechanism of inorganic carbon acquisition in lower brown algae

Stimulation or light-saturated rates of photosynthesis in Ectocarpus siliculosus (Dillwyn) Lyngb. by blue light was eliminated by increasing dissolved inorganic carbon (DIC) or by lowering pH in natural seawater. The amplitude of the circadian rhythm of photosynthesis was also diminished under these conditions, and the pH compensation points in a closed system were higher in the presence of blue light and during the circadian day. These observations suggest that blue light and the circadian clock regulate the activity of a carbon acquisition system in these plants. The inhibitor of external carbonic anhydrase, acetazolamide, reduced overall rates of photosynthesis by only about 30%, but ethoxyzolamide suppressed the circadian rhythm of photosynthesis almost completely and markedly reduced the duration of responses to blue light pulses. Similar patterns were obtained when photosynthesis was measured in strongly limiting DIC concentrations (0–0.5 mol m^?3). Since blue light stimulated photosynthesis under these conditions of strong carbon limitation, we suggest that blue light activates the release of CO₂ from an internal CO₂ store. We propose a metabolic pathway with similarities to that of CAM plants. Non-photosynthetic fixation leads to the accumulation of a storage metabolite. The circadian clock and blue light control the mobilization of CO₂ at the site of decarboxylation of this metabolite. In the presence of continuous blue light the pathway is proposed to cycle and act as a pump for CO₂ into the chloroplasts. This hypothesis helps to explain a number of previously reported peculiarities of brown algal photosynthesis.     

A new principle photosynthesis capacity biosensor based on quantitative measurement of delayed fluorescence in vivo

Delayed fluorescence (DF) is an excellent marker for evaluating plant photosynthesis. Compared with common methods for measuring the photosynthesis rate based on consumption of CO₂, DF technique can quantify the plant photosynthesis capacity more accurately and faster under its physiological status with less interference from the environment. We previously reported a method for measuring photosynthesis using DF of chloroplast [Wang, C.L., Xing, D., Chen, Q., 2004. Biosens. Bioelectron. 20, 454–459]. In the study, a novel fast and portable photosynthesis capacity biosensor system was developed, which was composed of light-emitting diode lattice as excitation light source, Channel Photomultiplier DC-Module to achieve DF, single-chip microcomputer as control center, hermetic dark sample chamber, battery power supply and CO₂, humidity and temperature controller. Compared with our previous work, the system was portable and can directly measure plant photosynthesis capacity in vivo in less than 10 s. A database in the software to carry out data acquisition and processing was developed to translate maximal DF intensity to net photosynthesis rate (Pn). In addition, local-control and remote-control mode can be chosen in the system. To demonstrate the utility of the system, it was applied to evaluate maximum Pn of four different plant species samples (Queen Rape Myrtle (var. rubra), soybean (Lu Hei No. 1), maize (Jin Dan No. 39) and rice (Jing Dao No. 21)) in field. The results were compared with that using commercial photosynthesis system LI-6400 and the uncertainty was less than ±5%. The new principle of photosynthesis measurement is a challenge and breakthrough to conventional method of gas exchange and may be a potential technique of next generation photosynthesis measurement.     

Endogenous isoprene protects Phragmites australis leaves against singlet oxygen

The possible protective role of endogenous isoprene against oxidative stress caused by singlet oxygen (¹O₂) was studied in the isoprene‐emitting plant Phragmites australis. Leaves emitting isoprene and leaves in which isoprene synthesis was inhibited by fosmidomycin were exposed to increasing concentrations of ¹O₂ generated by Rose Bengal (RB) sensitizer at different light intensities. In isoprene‐emitting leaves, photosynthesis and H₂O₂ and malonyldialdehyde (MDA) contents were not affected by low to moderate ¹O₂ concentrations generated at light intensities of 800 and 1240 µmol m^?2 s^?1, but symptoms of damage and reactive oxygen accumulation started to be observed when high levels of ¹O₂ were generated by very high light intensity (1810 µmol m^?2 s^?1). A dramatic decrease in photosynthetic performance and an increase in H₂O₂ and MDA levels were measured in isoprene‐inhibited RB‐fed leaves, but photosynthesis was not significantly inhibited in leaves in which the isoprene leaf pool was reconstituted by fumigating exogenous isoprene. The inhibition of photosynthesis in isoprene‐inhibited leaves was linearly associated with the light intensity and with the consequently formed ¹O₂. Hence, physiological levels of endogenous isoprene may supply protection against ¹O₂. The protection mechanisms may involve a direct reaction of isoprene with ¹O₂. Moreover, as it is a small lipophilic molecule, it may assist hydrophobic interactions in membranes, resulting in their stabilization. The isoprene‐conjugated double bond structure may also quench ¹O₂ by facilitating energy transfer and heat dissipation. This action is typical of other isoprenoids, but we speculate that isoprene may provide a more dynamic protection mechanism as it is synthesized promptly when high light intensity produces ¹O₂.     

树干皮层光合作用--生理生态功能和测定方法

大部分植物的树干(枝条)等部位含有能进行光合作用的绿色组织,树皮叶绿素含量最高可达750 mg/m2。这些绿色组织能够再固定树干内部的CO2(来源于自身组织呼吸或者木质部液流运输),使树干向大气排放的CO2量减少60%—90%皮层光合作用是树干生理活动的重要组成部分,其与树干呼吸和液流速率之间均有密切关系,对植物的碳平衡有重要作用。概述了皮层光合作用的生理生态功能;介绍了皮层光合作用测定和计算方法;讨论了皮层光合作用研究存在的问题;通过加入皮层光合作用的测量修正质量平衡法,以减少树干呼吸测定的不确定性。建议综合运用稳定碳同位素示踪、CO2和O2微传感器、树干液流技术等,准确地区分树干内部CO2的来源及比例,分析各个组分与影响因素的关系。同时,在微观上揭示皮层光合作用的基因组调控功能,在宏观上探讨尺度扩展、模型模拟,并与涡度协方差技术和遥感技术相融合以提高区域尺度估算的精度。     

Regulation of the mechanism for HCO 3 − use by the inorganic carbon level in Porphyra leucosticta Thur. in Le Jolis (Rhodophyta)

The capacity for HCO₃ ⁻ use by Porphyra leucosticta Thur. in Le Jolis grown at different concentrations of inorganic carbon (C_i) was investigated. The use of HCO₃ ⁻ at alkaline pH by P. leucosticta was␣demonstrated by comparing the O₂ evolution rates measured with the O₂ evolution rates theoretically supported by the CO₂ spontaneously formed from HCO₃ ⁻ . Both external and internal carbonic anhydrase (CA; EC 4.2.1.1) were implied in HCO₃ ⁻ use during photosynthesis because O₂ evolution rates and the increasing pH during photosynthesis were inhibited in the presence of azetazolamide and ethoxyzolamide (inhibitors for external and total CA respectively). Both external and internal CA were regulated by the C_i level at which the algae were grown. A high C_i level produced a reduction in total CA activity and a low C_i level produced an increase in total CA activity. In contrast, external CA was increased at low C_i although it was not affected at high C_i . Parallel to the reduction in total CA activity at high C_i is a reduction in the affinity for C_i, as estimated from photosynthesis versus C_i curves, was found. However, there was no evident relationship between external CA activity and the capacity for HCO₃ ⁻ use because the presence of external CA became redundant when P. leucosticta was cultivated at high C_i. Our results suggest that the system for HCO₃ ⁻ use in P. leucosticta is composed of different elements that can be activated or inactivated separately. Two complementary hypotheses are postulated: (i) internal CA is an absolute requirement for a functioning C_i-accumulation mechanism; (ii) there is a CO₂ transporter that works in association with external CA. Received: 20 April 1996 / Accepted: 5 August 1996