Fatty acid synthesis by spinach chloroplasts I. Property of fatty acid synthesis from acetate

Intact chloroplasts (about 70% Class I chloroplasts) isolatedfrom spinach leaves incorporated 150 nmoles of [1-¹⁴C] acetateinto fatty acids per mg chlorophyll in 1 hr at pH 8.3, 25°Cand 25,000 lux. On electron and phase-contrast microscopiescombined with hypotonic treatment of chloroplasts, this syntheticactivity was shown to be proportional to the percentage of ClassI chloroplasts in the preparation. Light was necessary for thesynthesis, the activity in the complete reaction mixture inthe dark being only 2% of that in the light. The synthetic activityincreased with increasing intensities of light to reach saturationat 6,000 lux. CoA and ATP were most effective as cofactors,HCO₃^–, HPO₄^2–, Mg^2$ and Mn^2$ were less effective.ATP could be replaced by ADP in the presence of Pi, suggestingpossible supply of ATP by photophosphorylation. Omission ofthe NADPH-generation system and NADH did not affect the synthesis,indicating sufficient provision of endogenous NADPH and NADHin intact chloroplasts under light. Addition of DTE did notcause recovery of the synthetic activity of intact chloroplastsin the dark. ¹ Present address: Radioisotope Centre, University of Tokyo,Yayoi, Bunkyo, Tokyo 113, Japan. (Received August 26, 1974; )     

Minor carotenoid components in the chloroplasts of higher plants

One-dimensional thin layer chromatography with microcrystallinecellulose was used for the separation of minor carotenoid componentsin spinach, parsley and Brassicachloroplasts. It was revealedthat chloroplasts of these plants contain two minor xanthophyllcomponents besides carotenes, lutein, violaxanthin and neoxanthin.These minor components, designated as xanthophyll–443and –439, were different in spectral properties from antheraxanthinand zeaxanthin which are known to be present in the chloroplastsof some higher plants, and evidence was obtained showing thatxanthophyll–439 has an epoxy group. Spectral propertiesof these xanthophylls in benzene, ethanol, H-hexane and carbondisulfide, as well as their contents relative to other carotenoids,are presented in this paper. ¹Postal address     

Cytoplasmic Alkalization and Cytoplasmic Streaming Induced by Light and Histidine in Leaf Cells of Egeria densa: in vivo 31P-NMR study

Rotational streaming of the cytoplasm including chloroplastswas induced by L-histidine, as well as by light, on the anticlinalface of leaf cells of Egeria densa. In the case of treatmentwith L-histidine some of the chloroplasts remained stationaryon the periclinal face of cells after rotational cytoplasmicstreaming was initiated. However, these chloroplasts were easilydislodged and translocated to the centrifugal end of the histidine-treatedcells by application of a centrifugal force that barely affectedthe location of chloroplasts in cells incubated in the darkwithout L-histidine. This result indicates that the anchoringof chloroplasts was weakened by L-histidine. Thus only the releaseof chloroplasts from anchoring was not enough for initiationof their streaming. The cytoplasmic pH (pH_c) and vacuolar pH(pH_v) were noninvasively monitored by in vivo ³¹P-nuclear magneticresonance (NMR) spectroscopy. Compared with the dark controlvalue, both illumination and treatment with L-histidine increasedthe pH_c by 0.3 units. In contrast, pH_v changed only a littlewith both illumination and treatment with L-histidine. Releaseof chloroplasts from anchoring and initiation of cytoplasmicstreaming are discussed in relation to the increase in pH_c inducedby both light and L-histidine. ⁴ Present address: Department of Cell Biology, National Instituteof Agrobiological Resources, Kannondai, Tsukuba, Ibaraki, 305Japan ⁵ Present address: Marine Biotechnology Institute Co., Ltd.,Head Office, 2-35-10 Hongo, Bunkyo-ku, Tokyo, 113 Japan (Received July 16, 1990; Accepted December 20, 1990)     

Analysis of Lipids in Prochloron sp.: Occurrence of Monoglucosyl Diacylglycerol

Prochloron contained monogalactosyl diacylglycerol, digalactosyldiacylglycerol, sulfoquinovosyl diacylglycerol, phosphatidylglyceroland, as a minor component, monoglucosyl diacylglycerol, butno phosphatidylcholine. With respect to the lipid and fattyacid compositions, this alga is similar to the blue-green algaerather than the chloroplasts of eukaryotic plants. ¹ Present address: Department of Biology, Faculty of Science,University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113, Japan. (Received August 24, 1982; Accepted November 18, 1982)     

Light-induced changes in membrane potential in Spirogyra

Spirogyra cells exhibited changes in membrane potential whenthey were exposed to light. Cells made chloroplast-free didnot show any light-induced potential change (LPC) upon illuminationwith white light and also monochromatic red (680 nm) and farred (720 nm) light. LPC was observed when the cell containedonly a small fragment of chloroplast, whether the cell had anucleus or not. The magnitude of LPC depended on the amountof chloroplast in the cell. DCMU at 10^–5 M, CCCP at 10^–5 M and DNP at 10^–4M at pH 5.5 suppressed LPC, while CCCP at 1–5 ? 10^–6M, NH₄Cl at 5 ? 10^–2 M and DNP at 10^–4 M at pH 7.0stimulated LPC. PMS at 10^–4 M stimulated LPC and couldinduce LPC which was completely inhibited by DCMU. These factssuggest that LPC is related to noncyclic and cyclic electronflows. The influences of light and dark conditions and various metabolicinhibitors (DCMU, DNP, CCCP, NH₄Cl) on ATP level have been investigated.No significant difference in the ATP level was observed betweencells in the light and dark. DNP at 10^–4 M (pH 5.5) andCCCP at 5 ? 10^–6 M decreased the ATP level significantly,while DCMU and NH₄Cl only slightly. Good correlation was notfound between the total ATP level and LPC in Spirogyra. LPC occurred even when the external medium contained only asingle salt such as KCl, NaCl or CaSO₄. LPC was also recorded in chloroplasts in situ and in vitro.The mode of LPC of chloroplasts was quite different from thatof the cell. On illumination, the chloroplast potential changedvery rapidly and transiently in the positive direction thenrecovered spontaneously to almost the original potential level. Possible causes of LPC are discussed in relation to the electrogenicion pump. ¹ Present address: Department of Botany, Faculty of Science,University of Tokyo, Hongo, Bunkyo, Tokyo 113, Japan. (Received November 9, 1977; )     

SOME PROPERTIES OF THE CYTOCHROME C REDUCING SUBSTANCE, A FACTOR FOR LIGHT-INDUCED REDOX REACTION OF CYTOCHROME C IN PHOTOSYNTHETIC LAMELLAE

Cytochrome c reducing substance (CRS), a redox substance discoveredin photoreactive lamellar fragments, was purified by Sephadexcolumn chromatography. Chromatographic behaviours of CRS ofAnabaena and spinach were essentially the same. Purified CRSof Anabaena showed an absorption spectrum having one absorptionmaximum around 260 mµ. The absorption peak disappearedon addition of excess amount of borohydride. Similar absorptionchange on borohydride addition was observed with spinach CRSpreparation. Purified preparations of Anabaena and spinach CRS supportedphotophosphorylation in spinach broken chloroplasts. The phosphorylationwas found to couple the electron flow from water to molecularoxygen. ¹This work was supported by grant GM-11300 from the NationalInstitute of Health, U. S. A. ²Present address: Institute of Applied Microbiology, The Universityof Tokyo, Tokyo, Japan.     

Enzymic synthesis of floridean starch in a red alga, Serraticardia maxima

ADP-glucose: -l,4-glucan -4-glucosyltransferase was obtainedfrom a marine red alga Serraticardia maxima in a form boundwith floridean starch granules. The enzyme catalyzed the transferof glucosyl residue from ADP-glucose, UDP-glucose and GDP-glucoseto floridean starch added as a primer. ADP-glucose was the mostefficient glucosyl donor in the reaction. Maltose was producedby ß-amylolysis of the glucan synthesized by the algalenzyme. The optimum pH for enzyme activity was at 8.4. The enzymewas not obtained in a soluble form from either the chloroplastextract or the whole algal cell extract. Electron micrographsof algal cells revealed that floridean starch granules are localizedexclusively outside chloroplasts. Hence, it appears that mostof the synthetase is present outside chloroplasts. ¹ Contribution from the Shimoda Marine Biological Station, TokyoKyoiku University, No. 202. This work was supported by a Grant-in-Aidfor Cooperative Research from the Ministry of Education, Japan. ² Present address: Department of Biology, Faculty of Science,Science University of Tokyo, Kagurazaka, Tokyo 162, Japan. ³ Present address: Laboratory of Biology, Faculty of Science,Toho University, Narashino, Chiba 275, Japan. (Received May 25, 1970; )     

Spectral Properties of Chromophore-Containing Fragments Prepared from Pea Phytochrome by Limited Proteolysis

Five chromophore-containing fragments were prepared from peaphytochrome in PR form (monomer mol wt 114,000) by limited proteolysiswith trypsin, thermolysin or chymotrypsin, and their absorptionand circular dichroism (CD) spectra were determined. The fragmentsof mol wt 62,100 and 56,400 showed photoreversible transformationbetween P_R and P_FR like phytochrome. The smaller fragments ofmol wt 40,300, 39,000 and 33,000 showed an absorption maximumat 657–660 nm (P₆₆₀) which was transformed to a bleachedform (P_BL) after a brief red-light exposure. The phototransformationbetween P₆₆₀ and P_BL was repeatedly reversible. Both P₆₆₀ andP_BL showed a negative CD band in red region like P_R, in contrastwith P_FR which has a positive band in far-red region. The natureof a chromophore domain of phytochrome and spectral propertiesof P_BL are discussed. ¹This study is dedicated to the late Professor J. Ashida. ²Permanent address: Department of Botany, Faculty of Science,University of Tokyo, Hongo, Tokyo 113, Japan. (Received August 7, 1982; Accepted March 26, 1983)     

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; )     

Studies on Plastid-Nuclei (Nucleoids) in Nicotiana tabacum L. III. Isolation of Chloroplast-Nuclei from Mesophyll Protoplasts and Identification of Chloroplast DNA-Binding Proteins

We have developed a method of isolating morphologically intactchloroplast-nuclei (nucleoids) in large quantities from mesophyllprotoplasts of Nicotiana tabacum L. cv. Bright Yellow-2. The isolated chloroplast-nuclei (cp-nuclei) were dispersed bythe treatments with proteinase K, 2 M NaCl and 2 M KCL, whichsuggested that the cp-nuclei are compactly organized by an electrostaticinteraction between the chloroplast-DNA and some protein(s).However, the four proplastid DNA-binding proteins identifiedpreviously (Nemoto et al. 1988) were not found in the cp-nuclei,and a different set of DNA-binding proteins (mol wt: 35 kDa,28 kDa and 26 kDa) was detected in the cp-nuclei by a DNA-bindingassay. On the other hand, the chloroplast-DNA was not differentfrom the proplastid-DNA. These findings indicate that the cp-nuclei are constituted fromthe plastid-DNA and the chloroplast-specific DNA-binding proteins.This suggests that the DNA-binding proteins in proplastids aredynamically replaced with the chloroplast DNA-binding proteinsduring the differentiation of plastids from proplastids to chloroplasts.The change of DNA-binding proteins may be involved in the morphologicalchange of plastid-nuclei and/or the regulation of plastid-DNAreplication and gene expression during the differentiation processof plastids. ⁶Present address: Department of Biotechnology, Faculty of Technology,Tokyo University of Agriculture and Technology, Nakamachi, Koganei,Tokyo, 184 Japan ⁷Present address: Department of Biology, Faculty of Science,University of Tokyo, Hongo, Tokyo, 113 Japan (Received April 4, 1990; Accepted May 18, 1990)     

Photosynthetic Characteristics of Chloroplasts Isolated from Euglena gracilis Z Grown Photoautotrophically

Photosynthetically competent chloroplasts were isolated fromcells of Euglena gracilis Z grown photoautotrophically in 1.5%CO₂. The isolated chloroplasts were intact and substantiallyfree from cytosolic, mitochondrial and microbody materials.The effects of some compounds on the activity of photosynthetic¹⁴CO₂ fixation were examined. The optimal pH and sorbitol concentrationwere 8.0 and 0.33 M, respectively. The chloroplasts requireda high level of P, (5 to 20 mM) for the maximal rate of photosynthesis.They were insusceptible to 10 mM of free Mg²⁺. ATP, ADP andAMP at 1 to 5 mM notably stimulated photosynthesis, althoughhigh concentrations of AMP were unfavorable. In the assay mediumdeveloped for this study, the chloroplasts exhibited photosyntheticactivity of 120µmoles-mg^–1 Chl-h^–1 at 30?C. Chloroplasts could also be isolated from cells grown under ordinaryair. The rate of photosynthetic ¹⁴CO₂ fixation at 1 mM NaH^l4CO₃was higher in these chloroplasts than in those isolated fromcells grown in 1.5% CO₂, whereas at 10 mM NaH^l4CO₃, the ratesof the two types of chloroplasts were nearly the same. Theseresults suggest that the CO₂ concentration given during growthof the algal cells affects the affinity for dissolved inorganiccarbon at the chloroplast level. (Received March 30, 1987; Accepted August 17, 1987)     

Temperature dependence of chlorophyll {alpha} fluorescence in lettuce and spinach chloroplasts at sub-zero temperatures

The temperature dependence of chlorophyll fluorescence wasmeasured in spinach and lettuce chloroplasts at sub-zero temperaturesin the presence of 50% ethylene glycol. In the presence of 5mM Mg²⁺, a fluorescence maximum appeared at –31?C in boththe spinach and lettuce chloroplasts, while in the presenceof only 5 mM Na⁺ as cations the maximum shifted to –20?Cin the spinach chloroplasts and to –11?C in the lettucechloroplasts. Since the occurrence of a maximum in the temperatureversus fluorescence curve is an indication for the transitionof the physical phase of thylakoid membrane lipids between theliquid crystalline and the phase-separation state (16, 18),these findings suggest that the (major) phase transition ofmembrane lipids occurs at these low temperatures in chloroplastsof higher plants and also that the phase transition temperatureis markedly lowered by the presence of divalent cations. Ethylene glycol at a concentration of 50% had almost no effecton the temperature dependence of chlorophyll fluorescence ina lamellar membrane preparation of Anabaena variabilis. In awater suspension of dimyristoylphosphatidylcholine, the additionof ethylene glycol to 50% did not alter the characteristic featureof the temperature dependence of fluorescence of 1-anilinonaphthalene-8-sulfonate.These findings suggest that 50% ethylene glycol does not affectthe temperature of the transition of the physical phase of membranelipids. ¹ C.I.W.-D.P.B. Publication No. 592. ² Present Address: Department of Biophysics and Biochemistry,Faculty of Science, University of Tokyo, Hongo 113, Tokyo, Japan. (Received June 22, 1977; )     

STIMULATORY EFFECT OF CARBON DIOXIDE UPON THE HILL REACTION AS OBSERVED WTTH THE ADDITION OF CARBONIC ANHYDRASE TO REACTION MIXTURE

The stimulatory effect of CO₂ upon the HILL reaction by isolatedchloroplasts was observed with erythrocyte carbonic anhydraseas a supplementary agent for CO₂ deprivation. Addition of thisenzyme to the reaction media remarkably shortened the time requiredto obtain the maximal effect of CO₂ The degree of stimulationwas rather small (below 50 per cent) and varied depending onthe preparation of chloroplasts. In general, the effect wasgreater with broken chioroplasts than with whole chloroplasts.The lowering of light intensity diminished the CO₂-effect. ¹ Present address: Laboratory of Biological Chemistry, TokyoInstitute of Technology, Meguro-ku, Tokyo. (Received April 6, 1962; )     

Changes in Protein Content and in the Structure and Number of Chloroplasts during Leaf Senescence in Rice Seedlings

Two types of experiment were carried out to examine whetheror not the inactivation of photosynthesis is related to lossof chloroplasts during foliar senescence of rice seedlings.Levels of both soluble and insoluble leaf proteins decreasedduring senescence, the loss of the soluble proteins being fasterthan that of the insoluble ones. There was a good positive correlationbetween the rate of oxygen evolution and the level of solubleproteins. The inactivation of photosynthesis was also linearlyrelated to the loss of a major fraction of insoluble proteins.Thus, the loss of photosynthetic ability is ascribable to thedegradation of relevant proteins and enzymes during leaf senescence.Electron microscopy revealed that senescence caused the disorientationof the grana and stroma thylakoids, a decrease in the numberof starch granules, and an increase in the size and number ofplastoglobuli. Large grana consisting 20 to 30 thylakoids appearedin aged leaves. In addition to these changes in ultrastructure,there was a significant decrease in the size of chloroplasts.Furthermore, the number of chloroplasts in mesophyll cells wasalso notably reduced during senescence. Thus, the loss of leafproteins and inactivation of photosynthesis are both relatedto the decrease in the total mass of chloroplasts during senescenceof rice seedlings. ³Present address: Department of Botany, Faculty of Science,University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113 Japan. (Received January 4, 1989; Accepted April 19, 1989)     

Multiplication and Differentiation of Plastid Nucleoids during Development of Chloroplasts and Etioplasts from Proplastids in Triticum aestivum

The morphological changes of plastid nucleoids (pt nucleoids)in the shoot apex and along the axis of the leaf blade in Triticumaestivum L. cv. Asakaze were followed with fluorescence microscopyafter staining with 4'6-diamidino-2-phenylindole (DAPI) andquantified with supersensitive microspectrophotometry. Proplastidsin the shoot apex contained 1–10 spherical pt nucleoids.These pt nucleoids changed to a row of spherical and cup-shapedpt nucleoids in sausage-shaped plastids at the leaf base inboth dark and light conditions, in which active cell divisionwas observed. These structures have a higher copy number ofplastid DNA (pt DNA) (72–78 copies) compared to proplastidsin the shoot apex (32–45 copies) and, therefore, may reflectthat active pt DNA synthesis is in progression. In the dark,the cup-shaped pt nucleoids in the spherical etioplasts, whichoriginated from the sausage-shaped plastids, grew to form ring-shapedpt nucleoids. Each ring-shaped pt nucleoid is sub-divided intosmaller pt nucleoids. Under continuous illumination, similarmorphological changes of pt nucleoids occurred except for distributionof small pt nucleoids into young chloroplasts as well as inmature chloroplasts. However, pt nucleotids of leucoplasts inepidermal and vascular bundle sheath cells did not show conspicuouschanges along the axis of the leaf blade. The significance ofthese observations is discussed in relation to plastid differentiationand to the plastid division cycle. ⁴ Present address: Faculty of Science, University of Tokyo,Hongo, Bunkyo-ku, Tokyo, 113 (Received August 15, 1989; Accepted April 13, 1990)     

Anion-dependent changes of energy transfer in chloroplasts III. Evidence for inactivation of light-triggered adenosine triphosphatase in chloroplasts by chloride

Washing chloroplasts with a high concentration of Tris-Cl^- buffercaused Cl^- dependent inhibition of photophosphorylation, light-inducedpH rise and light-triggered Mg²⁺-dependent ATPase activity.The inhibition of these activities was largely prevented bythe presence of 10^–4 M ADP or ATP during Tris washing,especially that of Mg²⁺-ATPase activity. The results were interpretedas suggesting that the inactivation of light-triggered ATPaseactivity in chloroplasts by chloride is one of the causes ofthe uncoupling of chloroplasts with Tris washing. (Received April 30, 1974; )     

Chloroplast DNA in Expanding Spinach Leaves

The proportion of chloroplast DNA in total DNA from spinachleaves has been measured using the second order reassociationkinetics of a ³H-labelled chloroplast DNA probe in total DNAextracts. There was no significant difference between the proportionof chloroplast DNA in the basal and distal halves of 2 cm leavesand in the distal halves of 5, 8, and 10 cm leaves. The meanof all the observations was 21.1 ± 0.7%. There was littlechange in the average total DNA content of cells from any ofthe leaves but cells from larger leaves contained 130–170chloroplasts while cells from the basal half of 2 cm leavescontained about 20 chloroplasts which were smaller than thosefrom the larger leaves. Consequently the average number of copiesof the plastome per chloroplast in large leaves was about 30(5 x 10^–15 g DNA) and in the smaller chloroplasts in thebase of 2 cm leaves was 200 (32 x 10^–15 g DNA). Stainingwith the DNA fluorochrome 4, 6-diamidino-2 phenyl indole (DAPI)showed 10–15 plastid nucleoid areas in chloroplasts oflarger leaves, suggesting there are 2–3 copies of theplastome per plastid nucleoid.     

Intra-Leaf and Intracellular Gradients in Chloroplast Ultrastructure of Dorsiventral Leaves Illuminated from the Adaxial or Abaxial Side during their Development

The number of thylakoids per granum, which is smaller in sun-typechloroplasts than in shade-type chloroplasts, was counted forthe chloroplasts at various positions within the single leavesof Spinacia oleracea L. and Glycine max (L.) Merrill. The thylakoidnumber increased with depth from the adaxial surface, but thistrend was not evident within respective cells. Therefore, photosyntheticproperties of chloroplasts should be similar within a cell butdifferent among cell layers. The similarity within a cell maybe due to the nuclear control of the chloroplast developmentand/or to chloroplast movement along the cell walls. Illumination of the leaves of G. max from the abaxial side duringtheir expansion resulted in the complete inversion of the intra-leafgradient in the thylakoid number, indicating that the formationof the intra-leaf gradient in chloroplast properties is influencedby the intra-leaf light environment during the later phase ofleaf development. ¹presennt address: Department of Environmental Biology, ResearchSchool of Biological Sciences, Australian National University,P.O. Box 475, Canberra, A.C.T. 2601, Australia. (Received March 13, 1986; Accepted May 30, 1986)     

Morphological and physiological characterization of IAA-less-sensitive and IAA-sensitive phases in rooting of Azukia cuttings

In disbudded epicotyl cuttings taken from light grown 5-dayold Azukia angularis Phaseolus angularis) seedlings, all adventitiousrootlets appeared on the second day of incubation. No root primordiawere observed within the first 24 hr and no increase in thenumber of roots occurred after 48 hr. Puromycin (5.5?10^–5M), p-fluorophenylalanine (1?10^–3M),2-thiouracil (2.3?10^–4M) and 2,6-diaminopurine (2?10^–5M)inhibited rooting when applied to cuttings on the second day,but showed no inhibition when applied on the first day. Unlike these inhibitors, pyrithiamine (7.2?10^–5M) inhibitedrooting when it was applied to cuttings on the first day. A rooting promoting effect was observed with actinomycin D (2.4?10^–6M),2,4-dinitrophenol (3?10^–5M) and p-fluorophenylalanine(1?10^–4M) applied to the cuttings on the first day, whereasindoleacetic acid (1.7?10^–4M) showed its promoting effectmost effectively on the second day. ¹Contribution No. 17 from the Botanical Gardens, Faculty ofScience, University of Tokyo, Tokyo, Japan. (Received June 4, 1969; )     

Photoreduction of 18O2 and H218O2 with Concomitant Evolution of 16O2 in Intact Spinach Chloroplasts: Evidence for Scavenging of Hydrogen Peroxide by Peroxidase

Illuminated intact spinach chloroplasts decomposed one moleculeof H₂¹⁸O₂ which resulted in the evolution of a half moleculeof ¹⁶O₂, but little ¹⁸O₂. The chloroplasts showed the same rateof photoreduction of ¹⁸C₂ as that of the evolution of ¹⁶O₂ withoutaccumulation of H₂¹⁸O₂. These reactions were suppressed by DCMU,and also by several inhibitors of ascorbate peroxidase and dehydroascorbateand monodehydroascorbate reductases in chloroplasts. These observationsindicate that the hydrogen peroxide produced in chloroplastsis reduced to water by a peroxidase using a photoreductant asthe electron donor. The hydrogen peroxide scavenging systemof chloroplasts was inactivated if hydrogen peroxide was addedin the dark, but not if added during the light. (Received May 4, 1984; Accepted July 10, 1984)