光呼吸对光合过程中磷代谢的影响

与光呼吸受抑制的 2％O_2浓度下相比,在 21％O_2浓度下.离体甘薯叶细胞光合作用最适介质无机磷浓度较低.另外,在21％O_2浓度下,降低甘薯叶细胞介质 NaHCO_3浓度,叶细胞光下吸收介质~(32)Pi的量减少;降低完整菠菜叶绿体介质 NaHCO_3浓度,乙醇酸形成相对加强,而介质~(32)Pi掺入到有机磷化合物的量则相对减少.这些结果表明,有利于光呼吸的条件,可降低光合对外界Pi的需求量.     

大豆和玉米光呼吸的控制

本文根据RuDP(1,5-二磷酸核酮糖)羧化酶的动力学特性测定了大豆光合作用和光呼吸的相对速率。在该羧化酶上,CO_2和O2对RuDP进行着争夺。高浓度的CO_2和低浓度的O_2,有利于RuDP羧化为三磷酸甘油酸,因而促进光合作用。低浓度的CO_2和高浓度的O_2则有利于RuDP的加氧反应而成为P-乙醇酸,因而促进光呼吸。在玉米上,PEP(磷酸烯醇式丙酮酸)羧化酶-苹果酸酶偶起了增加维管束鞘细胞中CO_2浓度的作用。四碳植物的维管束鞘细胞是RuDP羧化酶存在的场所。玉米的RuDP羧化酶对O_2也敏感,所以在维管束鞘内提高CO_2的浓度就能促进光合而减弱光呼吸。由于在800ppmCO_2和21％O_2的标准大气条件下,光呼吸及其相伴随的氧对光合的抑制作用,会明显地抑制大豆的净光合,同时由于迄未证明光呼吸有何重要的生理功能,因而正在试图设法减少或消除大豆和其他三碳植物的光呼吸。     

断光后的急骤耗氧现象及其与光呼吸的关系(英文)

用氧电极研究叶圆片断光后的耗氧变化,观察到断光后急骤耗氧(rapid postilluminationoxygen consumption RPIOC)现象。在不更换反应液的情况下,随着连续测定次数的增加,烟草叶圆片和番薯叶肉细胞的RPIOC增强。烟草、水稻、番薯、木瓜、黄瓜的叶圆片和番薯叶肉细胞表现出RPIOC,而高粱、甘蔗和玉米则没有这种现象。烟草叶国片的RPIOC随以下因素变化而增强:氧浓度从0.077mmol/L 气相O_27％)增加到 0.230 mmol/L(气相O_2 21％)、光强从111W/m~2增加到350W/m~2、温度从20℃增加到40℃,NaHCO_3(pH7.8)浓度从1 mmol/L(气相CO_2 1188ppm)降低到0.05mmol/L(气相CO_260 ppm)。0.4 mmol/L的HPMS完全抑制烟草叶圆片的RPIOC;10mmol/L的环氧丙酸抑制烟草叶圆片的RPIOC 57.8％。因此认为RPIOC是光呼吸的另一种形式的表现,它对环境因子的变化与已知的影响光呼吸的因子具有一致的反应。     

关于光呼吸与光合作用关系的研究——Ⅱ 环境因素对光合作用的影响及其与光呼吸的关系

采用光强、光质、照光时间、温度、气体成分以及叶水势、光合抑制剂等多种因素处理小麦叶片,观察这些因素对光合作用影响及其与光呼吸关系。结果证明,在各种因素的影响下,光合作用与光呼吸变化始终保持平行关系,而且在多数情况下,二者始终保持25％左右的比值,并发现二者比值变化受温度的调节。 发现光合与光呼吸从照光初始达最大速率(饱和值),要经过一个“滞后期”,小麦在10～15℃下,大约需55分钟,水稻在30℃下约需30分钟。 光呼吸猝发现象受光强、照光时间及高O_2影响;提高光强、延长照先时间或提高O_2浓度都可提高猝发强度,猝发强度一般可达33～81×10~(-3)CO_2mg dm~(-2)min~(-1)     

不同磷营养水平对烟草叶片光合作用和光呼吸的影响

随着磷营养水平的提高,烟草叶片的CO_2补偿点下降、光合速率上升。光呼吸在缺磷时最高。用光呼吸抑制剂处理烟草叶片后,光合的最适磷浓度提高。当CO_2浓度为560μl/L时,缺磷的烟草叶片在从21％O_2转入2％O_2时出现光合振荡,表明光呼吸与磷营养有密切关系。光呼吸在形成乙醇酸时所释放的磷,有回补叶绿体进行光合作用所需的磷的作用。     

单核细胞促进人外周血淋巴细胞离体增殖的自由基机理

在人外周血淋巴细胞培养体系中加入不同数量的单核细胞共同培养,以~3H-TdR掺入法和细胞色素C还原法分别测定淋巴细胞增殖能力和培养介质中O_2~-浓度;加入NADPH氧化酶抑制剂DPI实验性地下调O_2~-浓度,观察淋巴细胞增殖能力的变化。结果显示加入单核细胞可增加培养介质中O_2~-浓度,进而促进淋巴细胞增殖,提示O_2~-浓度的升高是淋巴细胞和单核细胞共培养体系中淋巴细胞增殖的重要因素。     

不同磷营养水平对烟草叶片光合作用和光呼吸的影响(英文)

研究了不同磷营养水平(0,1/4,1/2,1,2 P)对烟草(Nicotiana rustica L.)叶片光合、光呼吸、乙醇酸合成和乙醇酸氧化酶活性的影响,结果如下; 光合强度在0～1P范围内随磷水平的提高而增高,但在2P水平中略为下降。光呼吸强度在1/4～2P范围内与光合强度有相同的变化趋势,但在磷水平为零时最高;光呼吸/光合比值亦在磷水平为零时最高,并随磷营养的增加而下降。 HPMS抑制乙醇酸氧化酶活性,乙醇酸的积累量随磷水平的变化与光呼吸有一致的趋势。 在加入FMN时,不同磷营养水平的烟草叶片中乙醇酸氧化酶的活性随磷水平的提高而下降;加入FMN对酶活性的促进作用亦随磷水平的提高而下降。 叶片无机磷、有机磷及总磷含量均随磷营养水平的提高而增加。 用不同浓度的磷酸盐溶液真空渗入在1P培养的烟草叶圆片,并在0.25,0.5,10mmol/LNaHCO_3溶液中测定光合和光呼吸,结果表明在0.25和 0.5mmol/L NaHCO_3中,光呼吸随磷浓度的增加而下降;在 10mmol/L中光呼吸完全受抑制。光合作用与磷浓度关系呈单峰曲线,随着NaHCO_3浓度的提高,其高峰位置向右移,即光合最适磷浓度增大。 根据试验结果及从化学计量学推算,认为磷营养有抑制光呼吸的作用,而光呼吸的运转则有补充叶绿体内进行光合作用所需的无机磷的功能。     

神经节苷脂GM_3对人单核样白血病J6-2细胞中磷脂代谢的影响

用50μmol/L GM_3处理J6-2细胞6天,经组化与细胞学检查,证实细胞沿单核巨噬细胞途径分化。以[~(32)P]Pi或[CH_3-~(14)C]胆碱冲激(Pulse)后,将洗净的细胞进行Folch分配。取下相进行薄层层析,再做放射自显影。结果表明:GM_3的处理能促进[~(32)P]Pi或[CH_3-~(14)C]胆碱向磷脂酰胆碱的参入,而抑制向其他磷脂组分的参入。用[CH_3-~(14)C]胆碱冲激的Folch分配上相含水溶性胆碱代谢物,经TLC,结果表明CM_3的处理使[CH_3-~(14)C]胆碱向CDP-胆碱的参入减少。本研究证实,GM_3能调节J6-2细胞的磷脂代谢,其调节机制值得研究。     

二氧化碳和臭氧浓度升高对四季竹矿质养分含量和运输的影响

为阐明CO_2和O_3浓度升高对竹子矿质养分含量和运输的影响,以2年生四季竹(Oligostachyum lubricum)为试材,采用开顶式气室(OTCs)设置了环境背景大气[CK,(40±5)nmol·mol~(-1) O_3,(360±20)μmol·mol~(-1) CO_2]、O_3浓度升高[EO,(100±10)nmol·mol~(-1) O_3,(360±20)μmol·mol~(-1) CO_2]、CO_2浓度升高[EC,(40±5)nmol·mol~(-1) O_3,(700±35)μmol·mol~(-1) CO_2]、CO_2和O_3浓度复合升高[EOEC,(100±10)nmol·mol~(-1) O_3,(700±35)μmol·mol~(-1) CO_2]4个处理,测定了竹叶、竹枝、竹秆和竹根的Na+、Fe(~(2+,3+)、Ca~(2+)和Mg~(2+)含量,并分析了矿质营养在器官间的转运情况。结果显示:与CK比较,EO处理显著降低了四季竹植株体内Na+和Fe(~(2+,3+)含量,特别是竹根和竹叶,同时也明显降低Na+和Fe(~(2+,3+)器官间的运输能力。EC处理显著降低了四季竹植株体内Na+含量,而未改变其他矿质元素含量,但其器官中的分配格局和运输能力发生变化,尤其是竹枝向竹叶运输Ca~(2+)和Mg~(2+)的能力增强。EOEC处理显著降低了四季竹植株体内Na+含量,但显著提高了Fe(~(2+,3+)、Ca~(2+)和Mg~(2+)含量及其向上运输能力。研究表明,O_3浓度升高降低了四季竹植株体内矿质养分含量和器官间养分运输能力,在一定程度上影响四季竹的正常生长;CO_2浓度升高通过提高Ca~(2+)和Mg~(2+)向光合器官叶片的运输能力,促进四季竹生长;CO_2和O_3浓度升高复合作用能够通过提高四季竹光合器官竹叶中Fe(~(2+,3+)、Ca~(2+)和Mg~(2+)含量及其向光合器官叶片的运输能力,以维持体内矿质养分元素的平衡,提高四季竹对高浓度CO_2和O_3浓度复合环境下的适应能力。     

外源ABA对大蒜试管苗玻璃化发生和抗氧化系统的影响

以大蒜品种‘二水早’试管苗为材料,从活性氧代谢的角度研究了外源ABA、H_2O_2和H_2O_2+ABA处理下的试管苗玻璃化率、活性氧积累与组织定位和抗氧化系统的响应特征,探讨ABA缓解试管苗玻璃化过程的机理。结果表明:(1)外源H_2O_2处理可诱导大蒜试管苗玻璃化发生,外源ABA处理下玻璃化率最低,可以缓解H_2O_2诱导的玻璃化的发生。(2)试管苗O_2~产生速率和H_2O_2含量在H_2O_2处理下最高,在ABA处理下最低;在添加H_2O_2的培养基中同时添加ABA能显著减少因外源H_2O_2处理引起的O_2~产生和H_2O_2积累。(3)试管苗CAT、POD和APX活性在外源H_2O_2处理前期(0~8d)均上升并显著高于对照,但其CAT、APX活性在处理后期(8~16d)下降,其同期POD活性也增加缓慢;各抗氧化酶的活性在外源ABA与H_2O_2+ABA处理前期(0~8d)均呈直线上升趋势,而它们在H_2O_2+ABA处理后期(8~16d)均显著高于H_2O_2处理。(4)各处理试管苗抗坏血酸和谷胱甘肽含量随处理时间先升高后降低,并以外源ABA处理下最高,外源H_2O_2处理下最低。(5)试管苗O_2~和H_2O_2产生部位主要在基部和叶尖,且外源ABA处理下组织中ROS的积累最少。(6)在ABA+H_2O_2处理下,大蒜试管苗内丙二醛含量和膜相对透性显著低于对照和H_2O_2处理。研究发现,外源ABA处理可有效降低大蒜试管苗的内源O_2~产生速率和H_2O_2含量,提高抗氧化酶活性和抗氧化物质含量,抑制活性氧在试管苗内的产生和运输,显著降低试管苗玻璃化率;外源ABA可通过增强大蒜试管苗抗氧化能力来抑制玻璃化发生。     

Effect of glycidate on glycolate formation and photosynthesis in isolated spinach chloroplasts

Glycidate (2,3-epoxypropionate) increased CO₂ photoassimilation in intact spinach (Spinacia oleracea L.) chloroplasts in the presence of various inhibitors of photosynthesis, including O₂, arsenite, azide, iodo-acetamide, and carbonylcyanide 3-chlorophenylhydrazone. Although the mechanism by which glycidate enhances photosynthesis is obscure, the stimulatory effect cannot be ascribed to either an inhibition of glycolate formation, a specific interaction with the O₂ inhibition of photosynthesis, or a direct effect on the ribulose 1,5-bisphosphate carboxylase (EC 4.1.1.39) reaction. The lack of a differential effect of glycidate on photosynthesis and glycolate formation in the isolated chloroplast was confirmed in whole leaf studies by the CO₂ compensation concentration assay. These results are at variance with the report that glycidate stimulates net photosynthesis in tobacco leaf disks by irreversibly inhibiting glycolate formation and thus photorespiration (Zelitch, I., 1974, Arch. Biochem. Biophys. 163: 367-377).     

Evidence for recycling of inorganic phosphate by wheat chloroplasts during photosynthesis at air levels of CO2 and O2

Phosphate recycling under photorespiratory conditions was investigated using intact wheat chloroplasts from Triticum aestivum (cv. Maris dove). A decline in the optimal Pi level needed to support steady-state photosynthesis was observed (a) as the bicarbonate supply became limiting, or (b) as oxygen concentrations were increased. Further, at subsaturating CO2 and elevated O2 (52%), photosynthetic induction periods were shortest in the absence of exogenous Pi, and severely extended by its addition. Thus, photosynthesis under low CO2 levels which favor ribulose 1,5 bisphosphate (RuBP) oxygenase activity and glycolate synthesis by chloroplasts decreases their dependency on exogenous Pi from the initial illumination of chloroplasts through to the attainment of steady state rates of O2 evolution. Uptake of phosphate (Pi) was directly measured at ambient O2 concentrations and showed the stoichiometry of O2 evolved to Pi consumed at 10 mmol/L bicarbonate (saturating) had a mean value of 3.0, and was increased to 5.4 at 2.5 mmol/L bicarbonate and to > 8.0 at 1.0 mmol/L bicarbonate. The observation is consistent with enhanced stromal recycling of Pi released during hydrolysis of phosphoglycolate produced in greater quantities as the ratio of RuBP carboxylase relative to oxygenase activities (vc/vo) declines. The theoretical relationship between vc/vo and O2/Pi stoichiometries was derived and compared favorably to experimental data obtained.     

The stoichiometry of photorespiration during C3-photosynthesis is not fixed: evidence from combined physical and stereochemical methods

The stoichiometry of photorespiration, S, is defined as the fraction of glycolate carbon photorespired. It is postulated that under steady-state conditions there are two determinants of the ratio of photorespiration to net photosynthesis: the partitioning of ribulose bisphosphate between oxidation and carboxylation, and the partitioning of glycolate between reactions leading to complete oxidation to CO2 (S = 100%) and those yielding CO2 plus serine (S = 25%). S may be calculated using two independent probes of the system. The physical probe, using an infrared gas analyzer, measured photorespiration and net photosynthesis, and hence their ratio PR/NPS = pn(phys). The metabolic probe employed tracer (3R)-D-[3-3H1,3-14C]glyceric acid to determine r, the fraction of 3H retained in the triose phosphates leaving the chloroplasts. It is deduced from the postulated model that S = pn(phys) . r/(1 - r). Experiments have been performed with illuminated tobacco leaf discs (inverted) under varying concentrations of O2 and CO2. Increasing O2 at constant CO2 increased pn(phys) and decreased r, whereas increasing CO2 at constant O2 had the opposite effect. S more than doubled at 32 degrees C on going from 16 to 40% O2 (340 microliters CO2/liter) and decreased 40% on going from 200 to 700 microliters CO2/liter (21% O2). For discs in normal air S was somewhat greater than 27%. It is suggested that net photosynthesis, and therefore crop yields, could be increased by selecting for crop plants with reduced photorespiration stoichiometry.     

Chloroplastic photorespiratory bypass increases photosynthesis and biomass production in Arabidopsis thaliana

We introduced the Escherichia coli glycolate catabolic pathway into Arabidopsis thaliana chloroplasts to reduce the loss of fixed carbon and nitrogen that occurs in C(3) plants when phosphoglycolate, an inevitable by-product of photosynthesis, is recycled by photorespiration. Using step-wise nuclear transformation with five chloroplast-targeted bacterial genes encoding glycolate dehydrogenase, glyoxylate carboligase and tartronic semialdehyde reductase, we generated plants in which chloroplastic glycolate is converted directly to glycerate. This reduces, but does not eliminate, flux of photorespiratory metabolites through peroxisomes and mitochondria. Transgenic plants grew faster, produced more shoot and root biomass, and contained more soluble sugars, reflecting reduced photorespiration and enhanced photosynthesis that correlated with an increased chloroplastic CO(2) concentration in the vicinity of ribulose-1,5-bisphosphate carboxylase/oxygenase. These effects are evident after overexpression of the three subunits of glycolate dehydrogenase, but enhanced by introducing the complete bacterial glycolate catabolic pathway. Diverting chloroplastic glycolate from photorespiration may improve the productivity of crops with C(3) photosynthesis.     

Influence of Varying CO(2) and Orthophosphate Concentrations on Rates of Photosynthesis, and Synthesis of Glycolate and Dihydroxyacetone Phosphate by Wheat Chloroplasts

Intact chloroplasts of wheat (Triticum aestivum) were isolated from mesophyll protoplasts. With decreasing concentrations of bicarbonate from 10 to 0.3 millimolar (pH 8.0), the optimal concentration of orthophosphate (Pi) for photosynthetic O₂ evolution decreased from a value of 0.1 to 0.2 millimolar to 0 to 0.025 millimolar. The extremely low Pi optimum for photosynthesis at the low bicarbonate levels of 0.3 millimolar was increased by lowering the O₂ concentration from 253 (21% gas phase) to 72 micromolar (6% gas phase). The relative amount of glycolate and dihydroxyacetone phosphate (DHAP) synthesized under high and low levels of bicarbonate and varying levels of Pi was determined. At low levels of bicarbonate, glycolate was the main product, whereas at high bicarbonate levels, DHAP was the main product. Most of the DHAP and glycolate was found in the extrachloroplastic fraction.     

Inhibition of the phosphate transporter during isolation of intact chloroplasts from leaves of sunflower

Chloroplasts isolated from spinach leaves by the mechanical method were intact and exhibited high rates of CO₂-dependent oxygen evolution whereas chloroplasts isolated from sunflower leaves by the same technique were also intact but showed only low rates of oxygen evolution. The rate of uptake of orthophosphate (Pi) from the suspending medium with sunflower chloroplasts was less than 20% of that in spinach chloroplasts. The apparent Km for Pi transport was lower in sunflower chloroplasts but uptake was competitively inhibited by 3-phosphoglycerate in chloroplasts from both species. Uptake of malate (via the dicarboxylate transporter) and of ATP (via the adenine nucleotide transporter) was also reduced in sunflower chloroplasts compared to spinach chloroplasts. The endogenous Pi content and total exchangeable phosphate pool of sunflower chloroplasts were less than half that in spinach chloroplasts.Addition of a number of possible protective agents to the grinding medium failed to prevent the loss of photosynthetic activity during mechanical isolation of sunflower chloroplasts. Grinding mixtures of spinach and sunflower leaves together indicated that spinach chloroplasts were not inhibited by the sunflower leaf extract. Chloroplasts isolated from sunflower leaves via protoplasts had high rates of CO₂-dependent oxygen evolution. The Vmax and Km for Pi uptake, endogenous Pi content and total exchangeable phosphate pool of chloroplasts isolated from sunflower protoplasts were all similar to spinach chloroplasts. It is concluded that inner envelope membrane proteins are damaged during mechanical isolation of sunflower chloroplasts. The decrease in activity of the phosphate transporter and loss of endogenous phosphate may contribute to the low rates of photosynthesis observed in chloroplasts isolated by the mechanical method from leaves of sunflower and possibly other species.Abbreviations PGA 3-phosphoglyceric acid     

31P NMR studies of spinach leaves and their chloroplasts

An experimental arrangement is described which enables high quality 31P NMR spectra of compressed spinach leaf pieces to be continuously recorded in which all the resonances observed (cytoplasmic and vacuolar Pi, glycerate-3-P, nucleotides) were sharp and well resolved. 31P NMR spectra obtained from intact chloroplasts showed a distinct peak of stromal Pi. An upfield shift of the stromal Pi resonance was associated with a decrease in the external Pi and vice versa. Nucleotides were largely invisible to NMR in intact chloroplasts, whereas the same nucleotides reappeared in a typical 31P NMR spectrum of an acid extract of intact chloroplasts. Perfusion of compressed spinach leaf pieces with a medium containing Pi triggered a dramatic increase in the vacuolar Pi over 12 h. Addition of choline to the Pi-free perfusate of compressed leaf pieces resulted in a steady accumulation of phosphorylcholine in the cytoplasmic compartment at the expense of cytoplasmic Pi. When a threshold of cytoplasmic Pi concentration was attained, Pi was drawn from the vacuole to sustain choline phosphorylation. In spinach leaves, the vacuole represents a potentially large Pi reservoir, and cycling of Pi through vacuolar influx (energy dependent) and efflux pathways is an efficient system that may provide for control over the cytosolic-free Pi and phosphorylated intermediate concentrations. 31P NMR spectra of neutralized perchloric acid extracts of spinach leaves showed well defined multipeak resonances (quadruplet) of intracellular phytate. The question of cytosolic Pi concentration in green cells is discussed.     

Influence of glycerate on photosynthesis by wheat chloroplasts

Glycerate was found to effect photosynthetic O2 evolution in wheat chloroplasts by its conversion to triose phosphate and by influencing the rate of photosynthesis through the reductive pentose phosphate pathway. In the absence of bicarbonate, the photosynthetic O2 evolution with glycerate was low (10 to 25 mumol mg chlorophyll-1 h-1), and only about 15% of the rate of bicarbonate-dependent O2 evolution under optimum conditions. This corresponds to a rate of glycerate conversion to triose phosphate of 20 to 50 mumol mg chlorophyll-1 h-1, which appears sufficient to accommodate flux through the glycolate pathway in vivo. Pi was required for this glycerate-dependent O2 evolution; rates remained relatively constant between 0.1 and 40 mM Pi, and proceeded with little lag upon illumination (less than 0.5 min). Evidence for O2 evolution due to glycerate conversion to triose phosphate could be conclusively demonstrated by addition of glycolaldehyde, an inhibitor of the regenerative phase of photosynthesis, which prevents CO2 fixation. The effect of glycerate on photosynthesis in the presence of bicarbonate was determined by measuring both photosynthetic O2 evolution and 14CO2 fixation at varying Pi concentrations. Low concentrations of glycerate (micro- to millimolar levels) prevented inhibition of photosynthesis by Pi. With 1 mM bicarbonate and pH 8.2, which is favorable for glycolate synthesis, maximum rates of photosynthesis were obtained at low Pi (25 microM), whereas strong inhibition of photosynthesis occurred at only 0.2 mM Pi. Addition of glycerate relieved the inhibition of photosynthesis by Pi, indicating the possible importance of glycerate metabolism in the chloroplast under photorespiratory conditions. The initiation of photosynthesis by glycerate at inhibitory Pi levels occurred with little reduction in the ratio of CO2 fixed/O2 evolved, and the main effect of glycerate was on carbon assimilation. While the basis for the beneficial effect of glycerate on CO2 assimilation under moderate to high Pi levels is uncertain, it may increase the concentration of 3-phosphoglycerate (PGA) in the chloroplast, and thus make conditions more favorable for induction of photosynthesis and reduction of PGA to triose phosphate.     

Fixation of O(2) during Photorespiration: Kinetic and Steady-State Studies of the Photorespiratory Carbon Oxidation Cycle with Intact Leaves and Isolated Chloroplasts of C(3) Plants

Mass spectrometric techniques were used to trace the incorporation of [¹⁸O]oxygen into metabolites of the photorespiratory pathway. Glycolate, glycine, and serine extracted from leaves of the C₃ plants, Spinacia oleracea L., Atriplex hastata, and Helianthus annuus which had been exposed to [¹⁸O]oxygen at the CO₂ compensation point were heavily labeled with ¹⁸O. In each case one, and only one of the carboxyl oxygens was labeled. The abundance of ¹⁸O in this oxygen of glycolate reached 50 to 70% of that of the oxygen provided after only 5 to 10 seconds exposure to [¹⁸O]oxygen. Glycine and serine attained the same final enrichment after 40 and 180 seconds, respectively. This confirms that glycine and serine are synthesized from glycolate.

The labeling of photorespiratory intermediates in intact leaves reached a mean of 59% of that of the oxygen provided in the feedings. This indicates that at least 59% of the glycolate photorespired is synthesized with the fixation of molecular oxygen. This estimate is certainly conservative owing to the dilution of labeled oxygen at the site of glycolate synthesis by photosynthetic oxygen. We examined the yield of ¹⁸O in glycolate synthesized in vitro by isolated intact spinach chloroplasts in a system which permitted direct sampling of the isotopic composition of the oxygen at the site of synthesis. The isotopic enrichment of glycolate from such experiments was 90 to 95% of that of the oxygen present during the incubation.

The carboxyl oxygens of 3-phosphoglycerate also became labeled with ¹⁸O in 20- and 40-minute feedings with [¹⁸O]oxygen to intact leaves at the CO₂ compensation point. Control experiments indicated that this label was probably due to direct synthesis of 3-phosphoglycerate from glycolate during photorespiration. The mean enrichment of 3-phosphoglycerate was 14 ± 4% of that of glycine or serine, its precursors of the photorespiratory pathway, in 10 separate feeding experiments. It is argued that this constant dilution of label indicates a constant stoichiometric balance between photorespiratory and photosynthetic sources of 3-phosphoglycerate at the CO₂ compensation point.

Oxygen uptake sufficient to account for about half of the rate of ¹⁸O fixation into glycine in the intact leaves was observed with intact spinach chloroplasts. Oxygen uptake and production by intact leaves at the CO₂ compensation point indicate about 1.9 oxygen exchanged per glycolate photorespired. The fixation of molecular oxygen into glycolate plus the peroxisomal oxidation of glycolate to glyoxylate and the mitochondrial conversion of glycine to serine can account for up to 1.75 oxygen taken up per glycolate.

These studies provide new evidence which supports the current formulation of the pathway of photorespiration and its relation to photosynthetic metabolism. The experiments described also suggest new approaches using stable isotope techniques to study the rate of photorespiration and the balance between photorespiration and photosynthesis in vivo.