Effects of growth temperature on photosynthetic carbon metabolism in green plants III. Differences in structure, photosynthetic activities and activities of ribulose diphosphate carboxylase and glycolate oxidase in leaves of wheat grown under varied temperatures

The rate of ferricyanide photoreduction in broken chloroplastsisolated from leaves of wheat acclimatized to a low temperature(mean temperature, 5–7?C) was similar to that in chloroplastsfrom wheat acclimatized to a high temperature (20–25?C). There was no practical difference in glycolate oxidase activityin leaf extracts of wheat plants grown at low and high temperatures.In contrast, the ribulose diphosphate carboxylase activity ofchloroplasts from low temperature sample was less than halfthat for the high temperature sample. Chloroplasts having a high rate of photosynthetic CO₂-fixationwere obtained from wheat acclimatized to a low temperature,whereas the CO₂-fixation activity in chloroplasts isolated fromhigh temperature-acclimatized wheat was very low. Electron microscopy revealed that chloroplasts in high temperature-acclimatizedwheat were ellipsoidal, electron dense and contained starchgranules. Those in low temperature-acclimatized leaves wereround and did not contain starch granule. ²Present address: Department of Botany, Faculty of Science,University of Tokyo, Tokyo, Japan (Received August 7, 1973; )     

Biosynthetic mechanism of glycolate in Chromatium IV. Glycolate-glycine transformation

The metabolic transformation of glycolate to glycine occurringin photosynthesizing cells of Chromatium was investigated bythe radioisotopic technique and by amino acid analysis. By analyzingthe distribution of radiocarbon upon feeding [1-¹⁴C] glycolate,[2-¹⁴C] glyoxylate and [1-¹⁴C] glycine to bacterial cells, itwas demonstrated that glycolate is converted to glycinc viaglyoxylate, and both glycolate and glycine are excreted extracellularly.Although the formation of serine was barely detected by theabove two techniques in both N₂ and O₂ atmospheres, it was foundthat ¹⁴CO₂ is evolved quite markedly from both [1-¹⁴C] glycolateand [1-¹⁴C] glycine fed to the Chromatium cells. Analyticalresults of transient changes in amino acid compositions underatmospheric changes of N₂O₂ and by the addition of exogenousglycolate in N₂ confirm the notion that glycolate is convertedto glycine. Acidic amino acids (glutamic acid and aspartic acid)appear to take part in glycine formation as amino donors. Theformation of glycine from glycolate in a N₂ atmosphere suggeststhat an unknown glycolate dehydrogenation reaction may operatein the overall process. ¹ This is paper XXXVII in the series ‘Structure and Functionof Chloroplast Proteins’. Paper XXXVI is ref. (5). Theresearch was supported in part by grants from the Ministry ofEducation of Japan (No. 111912), the Toray Science Foundation(Tokyo) and the Naito Science Foundation (Tokyo). (Received July 14, 1976; )     

Effects of growth temperture on photosynthetic carbon metabolism in green plants I. Photosynthetic activites of various plants acclimatized to varied temperatures

In wheat, pea, barley, broad bean and rape, the photosyntheticcapacities in the leaves of plants acclimatized to a highertemperature (20–25?C) were much higher than those in thesame plants grown at low outdoor temperatures in winter (meanairtemperature, 5–7?C). On transferring the cold-grown wheat plants (vats, 5–7?C)to the higher temperature (25/20?C), the Photosynthetic rateof laves remained unchanged for the first 8 hr, then graduallyincreased to attain after 16 hr a maximum level which was thesame as that in wheat plants grown at the higher temperature(growth box, 20–25?C). When intact wheat without soilwas taken from the cold winter field and the attached leaveswere placed in the chamber at 25?C, the Photosyntheic rate ofthe attached leaves increased sluggishly for 2 hr, then remarkablyincreased to attain the maximum level 9 hr afer transferralto the chamber. When the leaves detached from wheat grown inthe winter field were transferred to the assimilation chamberkept at 25?C, the rate started to increase without a lag periodand attained the maximum level within 3 hr. The optimum temperaturefor photosynthesis shifted in parallel with the rise of airtemperature under which the wheat had been grown. The slopes of increase in the photosynthetic rates with increasingintensity of illumination wwere practically the same in wheatgrown under different temperatures. It was found that the total amount of sucrose in wheat grownat low temperatures in the winter was higher than that in thosegrown at the high temperature. ¹Present address: Department of Botany, Faculty of Science,University of Tokyo, Tokyo, Japan. (Received July 19, 1973; )     

Effect of the age of tobacco leaves on photosynthesis and photorespiration

Relationships among the activities of enzymes related to photosynthesisand photorespiration, and ¹⁴CO₂ photosynthetic products wereinvestigated with individual tobacco leaves attached to thestalk from the bottom to the top. P-glycolate phosphatase ofthe chloroplasts and glycolate oxidase of the peroxisomes hadtheir maximum activities in the 25th leaf from the dicotyledons.Maximum photorespiration was similarly distributed. The highestratio of serine-¹⁴C to glycine-¹⁴C in the photosynthesates andmaximum glycolate formation were also observed in the 25th leaf.Glutamateglyoxylate aminotransferase, serine hydroxymethyltransferaseand glycine decarboxylase were more active in the upper leaves.RuDP carboxylase had nearly constant activity in all leaves,except for the youngest in which activity decreased. MaximumCO₂ photosynthesis and enzyme activity for the C₄ dicarboxylicacid cycle occurred in the upper, youngest leaf. Distributionof photosynthetic CO² fixation among the leaves did not coincidewith RuDP carboxylase activity. The photosynthetic capacityappeared to be better related to the distribution pattern forenzymes of the C₄ dicarboxylic acid pathway, i.e. PEP carboxylase,pyruvate Pi dikinase and 3-PGA phosphatase in the upper leaves.The results suggest that the C₄ dicarboxylic acid pathway participates,to some extent, in photosynthesis in young leaves of tobacco,a dicotyledonous plant. ¹This work was reported at the Annual Meeting (1970) of theJapanese Plant Physiologists in Kobe. ²The Central Research Institute, Japan Monopoly Corporation1-28-3, Nishishinagawa, Shinagawaku, Tokyo, 141 Japan. (Received November 2, 1972; )     

Stimulation of Malate Synthesis by Glycolate in Tomato Leaves in Relation to CO2 Concentration

The mechanism by which malate synthesis from CO2 is increasedunder low concentrations of CO₂ was investigated in C₃ plants.A number of metabolites were administered to illuminated tomatoleaves, and their effects on the incorporation of ¹⁴CO₂ intomalate were determined. Compared with water as a control, glycolate,glyoxylate, D,L-glycerate, glycine, phosphoglycolate and L-serineincreased malate synthesis by factors of 6.8, 3.8, 3.3, 2.5,2.3 and 2.2, respectively. The effect of exogenous glycolateon malate synthesis from CO₂ was dependent on its concentrationup to 100 mu, but was independent of ambient CO₂ concentration.The feeding of l-¹⁴C-glycolate in the light indicated that glycolatestimulated the carbon flow from CO₂ to malate. The analysis of the products of ¹⁴CO₂ fixation in illuminatedleaves supplied with glycolate showed increases in malate andsugar and decreases in serine and phosphate esters. However,this stimulated malate synthesis ceased when malonate was suppliedsimultaneously with glycolate. Treatment with glycolate didnot affect the dark ¹⁴CO₂-fixation, but increased the ¹⁴C-malatesynthesis, with a corresponding decrease in ¹⁴C-aspartate and14^C-glutamate. These results suggest that exogenous glycolateactivates malate dehydrogenase in leaves, and that the increasedglycolate formation at low CO₂ concentrations is associatedwith the increased malate synthesis from CO₂. (Received January 12, 1981; Accepted May 20, 1981)     

Comparison of carbon dioxide fixation and the fine structure in various assimilatory tissues of Amaranthus retroflexus L

The pattern for primary products of CO₂-fixation and the chloroplaststructure of Amaranthus retrqflexus L., a species which incorporatescarbon dioxide into C₄ dicarboxylic acids as the primary productof photosynthesis, were compared in various chlorophyll containingtissues,i.e., foliage leaves, stems, cotyledons and pale-greencallus induced from stem pith. Despite some morphological differencesin these assimilatory tissues, malate and aspartate were identifiedas the major compounds labelled during a 10 sec fixation of¹⁴CO₂ in all tissues. Whereas, aspartate was the major componentin C₄-dicarboxylic acids formed in foliage leaves, malate predominatedas the primary product in stems, cotyledons and the pale-greencallus. The percentage of ¹⁴C-radioactivity incorporated intoPGA and sugar-P esters increased and ¹⁴C-sucrose was detectedin the prolonged fixation of ¹⁴CO₂ in the light, not only infoliage leaves, but also in stems and cotyledons. ¹ This work was supported by a Grant for Scientific ResearchNo. 58813, from the Ministry of Education, Japan. ² Present address: Institute of Applied Microbiology, Universityof Tokyo, Tokyo, Japan. ³ Present address: Department of Biochemistry, University ofGeorgia, Athens 30601. Georgia, U. S. A. (Received July 10, 1971; )     

Photoinhibition under Atmospheric O2, the Activation State of RuBP Carboxylase and the Content of Photosynthetic Intermediates in Soybean and Wheat

Ward, D. A. and Drake, B. G. 1987. Photoinhibition under atmosphericO₂, the activation state of RuBP carboxylase and the contentof photosynthetic intermediates in soybean and wheat.—J.exp. Bot. 38: 1937–1948. Associations between photosynthesis, the activation state ofRuBP carboxylase and the contents of photosynthetic intermediateswere compared in soybean and wheat leaves before and after exposureto photoinhibitory treatments in the presence of atmosphericO₂. Exposing attached leaves to a supra-saturating irradiance(3 800 µmol quanta m^{– 2} s^–1) for 2 h in CO_2-freeair decreased carboxylation efficiency and the light-saturatedphotosynthetic rate in air by approximately 50%. Exposure tothe photoinhibitory treatment for periods in excess of 2 h didnot cause a further decrease of photosynthesis in soybean. Althoughphotosynthesis was reduced, the initial and total (fully-activated)activities of ribulose 1,5-bisphosphate carboxylase (RuBPCase)in leaf extracts were unaltered in each species by the photoinhibitorytreatment. This was true for leaves sampled under both air andat a rate-limiting intercellular CO₂ partial pressure (C_i) of75 µPa Pa^–1. The contents of ribulose l,5-bisphosphate(RuBP) and 3-phosphoglyceric acid (3-PGA) were reduced by thephotoinhibitory treatment in soybean leaves sampled in air andat a rate-limiting C_i, although the RuBP/3-PGA ratio was unaffected.The relative reduction of RuBP content in soybean leaves atrate-limiting C₁ was similar to the corresponding reductionof carboxylation efficiency. For wheat,the relative reductionof RuBP content at rate-limiting C_i (–19%) caused by thephotoinhibitory treatment was considerably less than the correspondingdecrease of carboxylation efficiency (–49%).The RuBP/3-PGAratio of wheat was also increased significantly by the photoinhibitorytreatment The significance of these observations to the regulationof CO₂-limited photosynthesis in leaves experiencing photoinhibitionunder atmospheric oxygen is discussed. Consideration is alsogiven to the previous contention that contemporary measurementsof initial activity in crude extracts may provide a spuriousindication of the amount of the enzyme-CO₂-Mg_{2 +} form of RuBPcarboxylase present in the leaf. Key words: Carboxylation efficiency, RuBP carboxylase, photoinhibition, RuBP, 3-PGA     

Control of Photosynthesis in Wheat by CO2, O2 and Light Intensity

The regulation of photosynthesis in wheat leaves under varyingO₂, CO₂, and light was studied by analyzing certain metabolitepools and enzyme activities. Under high light when the rateof photosynthesis was limited by low intercellular levels ofCO₂ (C₁) there was a high level of ribulose-1,5-bisphosphate(RuBP) (about 100 nmols per mg chlorophyll). As C, increased,there was a parallel decrease in the ratios of RuBP/3-phosphoglycerate(PGA) (from 0.18 to 0.08 under 21% O₂) and triose-phosphate/PGA(from 0.16 to 0.07 under 21% O₂). The results suggest carboxylationis limited at low C_i, and that there is high carboxylation andlimited assimilatory power at high C_i. As photosynthesis increasedwith increasing Jight intensity under atmospheric levels ofCO₂ the ratios of RuBP/PGA and triosephosphate/PGA remainednearly constant (near 0.12 to 0.13) suggesting there may bea coordinate regulation by light of the different phases ofthe cycle. There was increasing activation of ribulose 1,5-bisphosphatecarboxylase oxygenase (Rubisco) and fructose 1,6-bisphosphatase(FBPase) with increasing light intensity. The ways in whichthe light activation of the enzymes Rubisco and FBPase may regulatecarbon metabolism in the cycle are discussed. ¹ Current address: Biological Sciences Center, Desert ResearchInstitute, PO Box 60220, Reno, Nevada 89506, U.S.A. (Received March 24, 1987; Accepted June 23, 1987)     

Glycine as a substrate for photorespiration

Substrates for photorespiration were examined by feeding ¹⁴Clabeled compounds to tobacco and corn leaf segments and by measuring¹⁴CO₂ evolution in light and darkness. CO₂ release in the darkwas rapid, but in light CO₂ release was slow due to refixationby photosynthesis. Carboxyl labeled glycine was more rapidlydecarboxylated than were glyoxylate, glycolate or serine. Hydroxypyridinemethanesulfonate, an inhibitor of glycolate oxidase, blocked CO₂ releasefrom glycolate but not from glycine. Isonicotynyl hydrazideblocked CO₂ release from both glycine and glycolate. DCMU blockedphotosynthetic refixation of the released CO₂, consequentlythe rates of CO₂ release in light and dark were about equal.It was concluded that CO₂ release during photo-respiration camefrom the conversion of 2 molecules of glycine to one serineand one CO₂. ¹⁴CO₂ release from glycine-l-¹⁴C in the dark or with DCMU inlight can be used as an assay for photorespiration ability. CO₂ release from glycine and glycolate by corn leaf segmentsin the dark proceeded at the rate of that in normal tobaccoleaf. This result, together with other work on O₂ exchange andenzymatic analysis, indicates that corn and other plants docarry on photorespiration, but it is not manifested by CO₂ releasein light. A yellow tobacco mutant, Consolation 402, had high rates ofphotorespiration by the ¹⁴CO₂ assay, nearly half (or more) asmany peroxisomes as chloroplasts, and high rates of CO₂ releasefrom glycine-l-¹⁴C or glycolate-l-¹⁴C. A common tobacco, BrightYellow, had lower rates of photorespiration, fewer visible peroxisomes,and slower decarboxylation of glycine and glycolate. The amount of ¹⁴CO₂ release from glycine-l-¹⁴C or glycolate-l-¹⁴Cincreased only slightly when the temperature was raised from25 to 35°C. ¹Parts of this work were abstracted at the Annual Meeting (April,1969) of Japanese Society of Plant Physiologists, Kanazawa ²Department of Biochemistry, Michigan State University, EastLansing, Michigan, U.S.A. (Received September 3, 1969; )     

The Effect of 2-Chloro-4-ethylamino-6-isopropyl-amino-s-triazine (Atrazine) on Distribution of 14C-Compounds Following 14CO2 Fixation in Excised Kidney Bean Leaves

Excised leaves of kidney bean plants treated with various concentrationsof atrazine for different periods were allowed to fix ¹⁴CO₂CO₂ fixation was inhibited by atrazine. The ¹⁴C-labelling patternof atrazine-treated leaves resembled dark Co₂-fixation patterns.The carbon-I-carboxyl group of ¹⁴C-aspartic acid from atrazine-treatedand ‘dark’ leaves showed no significant differencesin total radioactivity. Although atrazine disrupted the photosyntheticapparatus, it seemed to have no effect on non-photosyntheticCO₂ fixation.     

Effect of Carbonic Anhydrase on the Activity of Ribulose Bisphosphate Carboxylase

At concentrations of CO₂ less than saturating, carbonic anhydrase(EC 4.2.1.1 [EC] ) stimulates the carboxylation of ribulose bisphosphatecatalysed by ribulose bisphosphale carboxylase (EC 4.1.1.3 [EC] .9)in vitro. This is not through any beneficial association ofthe two enzymes but is a consequence of the increased rate ofconversion of HCO₃^– ion to CO₂, the substrate for thecarboxylation. Carbonic anhydrase should always be includedin reaction mixtures used to determine the Michaelis constantof ribulose bisphosphate carboxylase for CO₂ where fixationof radioactive CO₂ into phosphoglycerate is the basis of rateestimation. The effect is to decrease the value obtained forthe Michaelis constant.     

PHOTOSYNTHESIS OF GLYCINE AND SERINE IN GREEN PLANTS

1. The effects of "carbonyl" reagents on the photosyntheticin-corporation of ¹⁴CO₂ into the assimilation products of tobaccoand spinach leaves were studied. The presence of "carbonyl"reagents causes an increase in the ratio of ¹⁴CO₂ incorporatedin glycine and a decrease in serine. The incorporation of ¹⁴Cfrom glycolate-1-¹⁴C and glycolaldehyde-2-¹⁴C into glycine andserine was also affected by "carbonyl" reagents, as in the caseof ¹⁴CO₂-experiment. 2. The feeding experiments of glycine-1-¹⁴C and serine-1-¹⁴Cin the presence and in the absence of "carbonyl" reagents revealedthat these reagents inhibit the conversion of glycine to serine. 3. The results obtained above, together with the effects ofthiols on ¹⁴CO₂ incorporation presented in this paper, supportthe assumption that glycine and serine are formed via glycolateand glyoxylate during photosynthesis in green plants. 4. Comparison of ¹⁴C incorporation in malate from ¹⁴CO₂, glycolate-1-¹⁴C,glycine-1-¹⁴C and serine-1-¹⁴C in the presence and in the absenceof "carbonyl" reagents suggested the occurrence of the pathwayof the malate formation via glycolate and glyoxylate, not passingthrough glycine and serine, during photosynthesis. ¹ A part of this paper was presented at the Symposium on "Nitrogenand Plant" by the Japanese Society of Plant Physiologists, inOctober, 1963 ² Present address: Radiation Center of Osaka Prefecture, Sakai,Osaka     

Distribution of radioactivity in carbon atoms of malic acid formed during light-enhanced dark 14CO2-fixation in maize leaves

Most of the radioactivity incorporated into malic acid duringlight-enhanced dark ¹⁴CO₂-fixation was found in C-4, supportingour conclusion that phosphoenolpyruvic acid serves as a primaryacceptor of ¹⁴CO₂ to form the C₄ acid. ¹This work was reported at the 13th Annual Meeting of JapaneseSociety of Plant Physiologists, April, 1972. (Received January 6, 1973; )     

CO2-Fixation, Glycolate Formation, and Enzyme Activities of Photorespiration and Photosynthesis during Greening and Senescence of Sunflower Cotyledons

Time-courses of ¹⁴CO₂-fixation and of enzyme activities involvedin photorespiration and photosynthesis were determined duringthe life span of cotyledons from sunflower seedlings (Helianthusannuus L.). Glycolate formation in vivo was estimated from theresults of combined labelling and inhibitor experiments. NADPH-glyceraldehyde-3-phosphatedehydrogenase, NADPH-glyoxylate reductase and chlorophyll werewell correlated with the time-course of ¹⁴CO₂-fixation (photosynthesis).There was, however, a considerable discrepancy between the developmentalsequence of photosynthesis and that of both ribulose-l,5-bisphosphatecarboxylase and glycolate oxidase. Furthermore, time-coursesof glycolate oxidase activity in vitro and of glycolate formationin vivo differed significantly. Therefore, the use of glycolateoxidase as a marker for the activity of photorespiration ingreening sunflower cotyledons may be questionable. Results from¹⁴CO₂-labelling experiments with cotyledons treated with theglycolate oxidase inhibitor 2-hydroxy butynoic acid suggestthat glycolate formation relative to CO₂-fixation is reducedin senescent cotyledons. Key words: Development, glycolate oxidase, photorespiration, ribulose-l,5-bisphosphate carboxylase, oxygenase     

Light-enhanced carbon dioxide fixation in isolated chloroplasts

Preillumination of leaves of spinach, soybean and maize in theabsence of CO₂ greatly enhanced the capacity for fixing CO₂in an immediately following dark period. Lightenhanced darkCO₂-fixation was further observed in isolated chloroplasts ofspinach and soybean. When isolated chloroplasts were illuminated,CO₂-fixing capacity in the subsequent dark period increasedrapidly at first and later more slowly attaining a stationaryvalue in about 20 min. When the light was turned off at thisstage, the capacity decreased very rapidly becoming zero inabout 10 min. The magnitude of the enhanced dark fixation andits decay in the dark were not influenced by the presence orabsence of atmospheric oxygen. In both leaves and isolated chloroplasts,no significant change in oxygen (21%) occurred in distributionpatterns of radioactivity in products fixed by photosynthetic,or light-enhanced, dark, ¹⁴CO₂-fixation. In preilluminated leaves¹⁴C was incorporated into sucrose in the subsequent dark period,indicating that the photosynthetic carbon reduction cycle isoperating in light-enhanced dark fixation in higher plants. ¹Present address: Noda Institute for Scientific Research, Noda,Chiba Prefecture (Received August 10, 1970; )     

Excretion of glycolate during synchronous culture of Ankistrodesmus braunii in the presence of disalicylidenepropanediamine or hydroxypyridinemethanesulfonate

The rate of excretion of glycolate by the unicellular greenalga Ankistrodesmus braunii changes during its life cycle. Itis high in the main growth phase during the light period witha maximum 6 hr after the start of illumination, and low duringthe period of cell division in the dark. The glycolate excretion is stimulated by DSPD and HPMS, whilethe total ¹⁴CO₂-fixation is inhibited by DSPD and enhanced byHPMS. Changes in the effects of DSPD and HPMS on glycolate excretionas well as on photosynthetic ¹⁴CO₂-fixation during the courseof the algal life cycle were followed using the technique ofsynchronous culture. How far the change of glycolate excretion is due to a changeof glycolate oxidase activity during the life cycle and to achange of C₂-supply from the carbon reduction cycle is discussed.The effect of DSPD on glycolate excretion suggests a participationof ferredoxin in the glycolate pathway. (Received August 10, 1968; )     

Photosynthetic and light-enhanced dark fixation of 14CO2 from the ambient atmosphere and 14C-bicarbonate infiltrated through vascular bundles in maize leaves

1. The capacity of light-enhanced dark fixation of ¹⁴CO₂ from theambient atmosphere decayed following time-course characteristicsof a first-order reaction (half-life, 1–2 min). The levelof phosphoenolpyruvate in maize leaves under CO₂-free air didnot decrease in the dark subsequent to preillumination. Theseresults indicate that phosphoenolpyruvate carboxylase is activatedin light and quickly inactivated in the following darkness.
2. Removal of oxygen from the atmosphere did not exert any effecton the products of light-enhanced dark fixation of ¹⁴CO₂ providedfrom the atmosphere, the major labeled compounds being malateand aspartate. This confirms that the transfer of carboxyl carbonof C₄-acids to form 3-phosphoglycerate is light-dependent.
3. WhenNaH¹⁴CO₃ solution was vacuum-infiltrated through vasculartissuesof maize leaves, the main initial photosynthetic ¹⁴CO₂fixationproducts were phosphate esters. This indicates thatby thistechnique, ¹⁴CO₂ could be directly provided to the bundlesheathcells, and was fixed via the reductive pentose phosphatecycle.On the other hand, the main initial ¹⁴CO₂-fixation productswere malate and aspartate even when ¹⁴CO₂ was provided throughvascular tissues in the dark immediately following preillumination.The possible regulatory mechanisms underlying the above findingsare discussed.

¹ This work was reported at the 4th International Congress onPhotosynthesis, Reading, September 1977. Request for reprintsshould be addressed to S. Miyachi, Institute of Applied Microbiology,University of Tokyo, Bunkyo-ku, Tokyo 113, Japan ² Present address: Okinawa Branch of Tropical Agriculture ResearchCenter, Ishigaki-shi, Okinawa 907, Japan. (Received October 28, 1977; )     

Biosynthetic mechanism of glycolate in Chromatium 6. Glycolate formation and metabolism under low O21

Under low O₂ (0.05 mM O₂), there was no measurable excretionof glycolate or glycine by Chromatium cells, unlike the caseof their incubation under high (0.7 mM) O₂ However, upon additionof non-radioactive glycolate and glycine to the suspension medium,there occurred a measurable incorporation of ¹⁴CO₂ into thesecompounds, which were then excreted extracellularly; the totalradioactivities measured were approximately 15% of the totalCO₂ fixed photosynthetically. This phenomenon could be as cribedto the dilution of the intracellular pools by the compoundsadded. The results indicate that under low O₂ the glycolatemolecules produced are metabolically further transformed inthe bacterial cells. The incorporation of ¹⁴CO₂ into the extracellularglycolate fraction was maximal at 0.3 mM glycolate in both highand low O₂. Presumably, glycolate formed in the bacterial cellsunder both the high and low O₂ is metabolized in a similar manner,although the excess glycolate and glycine molecules are rapidlyexcreted. During glycolate metabolism CO₂ was evolved from anisonicotinylhydrazide-sensitive reaction, suggesting that thepathway <glycolate glycine . CO₂ was similar in green plants.The results thus indicate that studies on glycolate and glycinemetabolism in the anaerobic bacterium, Chromatium, provide auseful model system for elucidating the mechanism of photorespirationin green plants. (Received May 19, 1978; )     

Photorespiration: stimulation of glycine decarboxylation by oxygen in tobacco leaf disks and corn leaf segments

Glycine decarboxylation, shown by us as an intermediate stepin CO₂ evolution during photorespiration, was enhanced by oxygentensions of up to 100%, whereas CO₂ evolution from glucose didnot increase when oxygen tensions were raised above 20%. Thus,we concluded that enhancement of photorespiration by oxygenis not only due to stimulation of glycolate oxidation, but alsoto stimulation of glycine decarboxylation. ¹ This work was reported at the Annual Meeting (1971) of theJapanese Society of Plant Physiologists in Okayama. ² The Okayama Tobacco Experiment Station, Japan Monopoly Corporation,Tamashima, Kurashiki, Okayama. ³ The Hatano Tobacco Experiment Station, Japan Monopoly Corporation,Naganuki, Hatano, Kanagawa. (Received October 19, 1971; )     

Biosynthetic mechanism of glycolate in Chromatium I. Glycolate pathway

The pattern of radioactivity distribution in several amino acidsof Chromatium cells exposed to ¹⁴CO₂ was determined. By transferringthe bacterial cells from an atmosphere of nitrogen to oxygenthere occurred a transient decrease of ¹⁴CO₂ incorporation intoaspartate and glutamate, whereas that into glycine showed aprominent increase. The labeling of both serine and alaninedid not show a marked change under such conditions. The, activitiesof glycolate oxidase and glycolate dehydrogenase in crude extractsof the bacterial cells were very low. The formation of glycolic acid only occurred during the oxidativemetabolism of Chromatium cells grown on bicarbonate as a C source,being negligibly small in bacteria under nitrogen or after growthon malate or acetate. The activities of both ribulose- 1,5-bisphosphateoxygenase and phosphoglycolate phosphatase in the extract preparedfrom the bicarbonate-grown bacterial cells were very low andapparently could not account for the glycolic acid formationthrough these enzymic reactions. Metabolic patterns of glycolicacid in Chromatium are discussed in relation to the photorespiratoryphenomenon. (Received February 24, 1975; )