Inhibition site of the electron transport system in lettuce chloroplasts by fumigation of leaves with SO2

In chloroplasts isolated from SO₂-fumigated leaves at 2.0 ppm,electron flow from water to 2,6-dichloroindophenol (DCIP) wasinhibited, but the electron flow from reduced DCIP to methylviologen was not affected. Neither diphenylcarbazide nor MnCl₂could restore the activity of the DCIP-Hill reaction of SO₂-injuredchloroplasts. Electron flows, from water to ferricyanide orto silicomolybdic acid, were inhibited in a degree similar tothat of the DCIP-Hill reaction. The rate of carotenoid photobleaching in the presence of carbonylcyanide-m-chlorophenylhydrazone was suppressed and paralleledthe inhibition of the DCIP-Hill reaction. In SO₂-injured chloroplasts, the variable part of the fluorescencetransient was diminished, and the fluorescence yield loweredby SO₂ was increased with 3-(3', 4'-dichlorophenyl)-l, l-dimethylurea(DCMU) or more pronouncedly by incubating the sample with sodiumdithionite. However, the yield could not recover to the levelfound in non-fumigated chloroplasts. With SO₂ fumigation, thetime required to reach steady-state level of fluorescence becamelonger in the absence of DCMU, but was not altered in the presenceof DCMU. The pool size of the primary electron acceptors decreasedwith SO₂ fumigation. We concluded that SO₂ inactivated the primaryelectron donor or the reaction center itself. The mode of SO₂action in the electron transport chain is discussed. (Received October 20, 1979; )     

Retardation of Leaf Senescence by Ascorbic Acid

Leaf discs of Solatium melongena were floated on various concentrationsof ascorbic acid (AA), gibberellic acid (GA₃), and kinetin inorder to study their effect on senescence. AA was highly effectivein retarding senescence as shown by the arrest of the fall inlevels of chlorophyll, DNA, RNA, and proteins. AA was effectiveat a lower concentration than that of GA₃ or kinetin.     

Ascorbic Acid-Dependent Regulation of Redox Levels of Chlorogenic Acid and its Isomers in the Apoplast of Leaves of Nicotiana tabacum L.

There is a question whether ascorbic acid (AA) can control redoxlevels of phenolics in the apoplast. The present study was designedto answer this question. AA, dehydroascorbic acid (DHA), chlorogenicacid (CGA) and its two structural isomers were present in theapoplast of leaves of tobacco (Nicotiana tabacum L. cv. BelW3).The levels of AA plus DHA (AA + DHA) and the ratios of AA to(AA + DHA) decreased while the levels of CGA plus its isomersincreased during leaf aging. o-Quinones of CGA plus its isomerswere found in the apoplast only in aged leaves of which apoplasticlevel of AA was nearly zero. In addition, activity of apoplasticperoxidase that could oxidize CGA and its isomers increasedduring leaf aging. From the observations, it is concluded thatAA can regulate the accumulation of the o-quinones of CGA andits isomers in the apoplast. Based on the conclusion, it isproposed that soluble peroxidase in the apoplast has two functions,namely, (i) scavenging of H₂O₂ and/or regulation of the levelof apoplastic H₂O₂ in the presence of AA, and (ii) accumulationof oxidation products of the phenolics in the absence of AA. (Received January 30, 1998; Accepted April 7, 1998)     

Inhibition of Photosynthesis by Sulfite in Mesophyll Protoplasts Isolated from Vicia faba L. in Relation to Intracellular Sulfite Accumulation

Inhibition of photosynthesis by Na₂SO₃ in mesophyll protoplastsisolated from Vicia faba leaves and uptake of sulfite by theprotoplasts were examined at various pH values of the incubationmedium containing Na₂SO₃. As the pH of the incubation mediumlowered, the rate of photosynthesis in the protoplasts decreasedand the amount of sulfite taken up by the protoplasts increased.Most of sulfite accumulated in the protoplasts was not metabolizedduring the dark incubation, as measured with an ion chromatograph.Photosynthetic O₂ evolution by the chloroplasts isolated fromVicia mesophyll protoplasts was inhibited by exogenously-appliedNa₂SO₃ over pH region examined (7.4–9.0). The sulfiteconcentration required for a half inhibition of photosynthesisby the isolated chloroplasts was similar to the intracellularsulfite level required for that by the protoplasts. These resultsindicate that the intracellular sulfite accumulated in the protoplastsin an unmetabolized state is responsible for the inhibitionof protoplast photosynthesis. (Received January 24, 1985; Accepted May 29, 1985)     

The Large Subunit of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase is Fragmented into 37-kDa and 16-kDa Polypeptides by Active Oxygen in the Lysates of Chloroplasts from Primary Leaves of Wheat

Lysates of chloroplasts isolated from wheat (Triticum aestivumL. cv. Aoba) leaves were incubated on ice (pH 5.7) for 0 to60 min in light (15 µmol quanta m^–2 s^–1),and degradation of the large subunit (LSU) of ribulose-l,5-bis-phosphatecarboxylase/oxygenase (Rubisco: EC 4.1.1.39 [EC] ) was analyzed byapplying immunoblotting with site-specific antibodies againstthe N-terminal, internal, and C-terminal amino acid sequencesof the LSU of wheat Rubisco. The most dominant product of thebreakdown of the LSU and that which was first to appear wasan apparent molecular mass of 37-kDa fragment containing theN-terminal region of the LSU. A 16-kDa fragment containing theC-terminal region of the LSU was concomitantly seen. This fragmentationof the LSU was inhibited in the presence of EDTA or 1,10-phenanthroline.The addition of active oxygen scavengers, catalase (for H₂O₂)and n-propyl gallate (for hydroxyl radical) to the lysates alsoinhibited the fragmentation. When the purified Rubisco fromwheat leaves was exposed to a hydroxyl radical-generating systemcomprising H₂O₂, FeSO₄ and ascorbic acid, the LSU was degradedin the same manner as observed in the chloroplast lysates. Theresults suggest that the large subunit of Rubisco was directlydegraded to the 37-kDa fragment containing the N-terminal regionand the 16-kDa fragment containing the C-terminal region ofthe LSU by active oxygen, probably the hydroxyl radical, generatedin the lysates of chloroplasts. (Received October 28, 1996; Accepted February 7, 1997)     

Proton Neutralization in the Leaves of English Oak (Quercus robur L.) Exposed to Sulphur Dioxide

Reactions to the input of acidic gases were investigated inleaves of Quercus robur L. exposed to different concentrationsof SO₂ (80, 120, and 160 nl I^–1) for 32 to 70 d. Two-year-oldoaks were grown in nutrient solutions with varied nitrogen formand were fumigated in closed chambers. An attempt was made toidentify the mechanisms of proton neutralization by consideringthe uptake of nitrogen, the increase in sulphur and carboxylatecontents, and the excretion of hydroxyl ions or protons. Inaddition, nitrate reductase activity was determined in the leaves. The reduction of sulphur was not involved in the neutralizationof protons generated by SO₂-uptake, whereas organic acid metabolismplayed a decisive role. Depending on SO₂-concentration, durationof fumigation and nitrogen supply, oaks reacted with a reductionin the size of the carboxylate pool in the leaves, and/or withan increase in proton excretion (or a decrease in hydroxyl ionexcretion). Nitrate reductase activity increased in the leavesof nitrate-grown oaks exposed to the highest SO₂-concentration(160 nl l^–1) for 42 d. The capacity of the mechanismsconsidered is sufficient for the neutralization of the calculatedamounts of protons resulting from SO₂-uptake. Key words: Leaves, neutralization, protons, Quercus, sulphur dioxide     

The Effect of Sulphur Dioxide on the Growth of Lolium perenne L., Lolium multiflorum Lam., Dactylis glomerata L., and Phleum pratense L.

Lockyer, D. R. 1985. The effect of sulphur dioxide on the growthof Lolium perenne L., Lolium multiflorum Lam., Dactylis glomerataL., and Phleum pratense L.?J. exp. Bot. 36: 1851-1859. Fouragriculturally important grasses, Lolium perenne L., Loliummultiflorum Lam., Dactylis glomerata L. and Phleum pratenseL. were exposed to sulphur dioxide (SO₂) in a system of exposurechambers. The plants were exposed for a total of 43 d to meanconcentrations of SO₂ in the air of 0,87 or 448 (µg m^–3and herbage was harvested twice. All four grasses showed chloroticlesions after exposure to the highest concentration of SO₂.The effect of SO₂ on the yield of herbage was statisticallysignificant only at the second harvest and at the highest concentration;the dry weights of shoots of D. glomerata and L. perenne werereduced by 33% and 16% respectively. Significant effects ofSO₂ were also found on the 'transpiration coefficients' measuredfor D. glomerata and P. pratense. The grasses differed in theiruptake of sulphur from the atmosphere but this was not relatedto their sensitivity to SO₂. Total–S concentration inthe shoots of L. perenne, L. multiflorum and D. glomerata increasedalmost linearly in response to increasing SO₂ concentration;with P. pratense only the highest SO₂ concentration raised total-Sabove the level in control plants. These increases were almostentirely due to the accumulation of sulphate–S. Key words: Sulphur diozide, Lolium perenne, Lolium multiflorum, Dactylis glomerata, Phleum pratense     

Inter-population Variations of Kochia indica During Germination Under Different Stresses

Germination of three populations of Kochia indica Wight fromcontrasting soil types were studied under various levels ofsalinity, alkalinity and osmotic stress with a view to evaluatetheir potential to establish through seeds in saline and alkalineenvironments of various magnitudes. The seeds were germinatedunder controlled conditions in Petri dishes using salinizedsolutions of NaCl, Na₂SO₄, MgCI₂ and CaSO₄ (electrical conductivity,EC, 4–16 ds m^–), sodium carbonate solutions (pH8.5–100), mannitol solutions ( – 2.5 to –100 bar) and de-ionised distilled water (control). Inter-populationdifferences were very marked, showing selective adaptation ofthe populations to a particular stress type. In general, allpopulations exhibited salt tolerance but were rather sensitiveto high osmotic stress. Kochia indica Wight (bui), germination, population, salinity, alkalinity, osmotic stress     

The Transport and Metabolism of Urea in Chara australis: II. UREASE AND THE DISTRIBUTION OF TRANSPORTED UREA

The ureolytic enzyme in Chara was investigated. This enzymewas shown to be a urease with an unusually high affinity forurea(K_m = 158 mmol m^-3). Little inhibition of urease activitywas found when intact Chara cells were exposed to the ureaseinhibitors hydroxyurea, acetohydroxamic acid and N-ethylmaleimide,although there was some inhibition of urea uptake. The distribution of radioactivity amongst the amino acid, organicacid and sugar/neutral fractions, determined by ion-exchangechromatography, was very similar whether the Chara internodeswere exposed to ¹⁴C-urea or to H¹⁴CO₃. This suggests that thefraction of the urea-carbon liberated by the urease as CO₂ andretained by the cell is used in photosynthetic carbon-fixation.During the initial 15 min of ¹⁴C-urea uptake, label appearsin the vacuole only in the form of unmetabolized urea. Afterthis time a variety of labelled compounds appear in the vacuole,presumably reflecting the gradual movement of carbon-fixationproducts from the chloroplasts to the cytoplasm and thence intothe vacuole. Key words: Urea transport, metabolism, Chara, urease     

Participation of Hydrogen Peroxide in the Inactivation of Calvin-Cycle SH Enzymes in SO2-Fumigated Spinach Leaves

In SO₂-fumigated spinach leaves under light, chloroplast SHenzymes, glyceraldehyde-3-phosphate dehydrogenase (NADP-GAPD)(EC 1.2.1.13 [EC] ), ribulose-5-phosphate kinase (Ru5PK) (EC 2.7.1.19 [EC] )and fructose-1,6-bisphosphatase (FBPase) (EC 3.1.3.11 [EC] ) weremore remarkably inactivated than other chloroplast enzymes.Their activities recovered after removal of SO₂. The inactivationparalleled light-dependent CO₂-fixation in spinach leaves. Inilluminated chloroplasts isolated from SO₂-fumigated spinachleaves, NADP-GAPD and Ru5PK were more specifically in activatedthan other chloroplast enzymes. These two enzymes could be protectedfrom the inactivation by adding catalase. The NADP-GAPD inactivationwas suppressed by DCMU, cytochrome c or anaerobic conditions.By adding thiol compounds, the NADP-GAPD inactivation was dischargedand the activity increased. In chloroplasts or crude extractsfrom non-fumigated spinach leaves, NADP-GAPD and Ru5PK weremore strongly inhibited by externally added H₂O₂ than otherchloroplast enzymes. All results supported the idea that thesuppression of photosynthesis at the beginning of SO₂ fumigationwas caused by the reversible inhibition of chloroplast SH enzymewith H₂O₂. (Received October 7, 1981; Accepted June 16, 1982)     

Changes in Growth and Quality Characteristics of Lucerne (Medicago sativa L.) in Response to Sulphur Dioxide Exposure under Field Conditions

The effects of SO₂ on some growth and quality characteristicsof lucerne (Medicago sativa L.) were investigated by exposingplants to mean SO₂ concentrations of 215, 78 or 2.8 µgm_–3 in open-top chambers for 166 d. Plants exposed to215 µg m_–3 had significantly lower shoot and rootweights compared with plants exposed to 78 µg m^–3,but not compared with control plants. Exposure to 215 or 78µg m ^–3 increased the plant shoot: root ratio, buthad no effect on leaf area. During the middle of the fumigationperiod, relative growth rate and net assimilation rate werehighest in plants exposed to 215 fig m, but these later fellbelow control values, and plants exposed to 78 µg m^–3had the highest relative growth rate and net assimilation rate.As the duration of exposure increased, an initial SO₂-inducedstimulation of growth may have developed to toxicity at thehighest SO₂ exposure. Exposure to SO₂ depressed L-ascorbic acid concentrations inleaves, had no effect on foliar protein or starch concentrations,and increased the specific energy of shoots and plant sulphurconcentrations. The effect of SO₂ on L-ascorbic acid concentrationsmay suggest a mechanism for reduced freezing tolerance of plantsafter exposure to SO₂. Key words: SO₂, Medicago sativa L., Growth     

Intracellular Localization of Phosphoenolpyruvate Carboxykinase in Bundle Sheath Cells of C4 Plants

Bundle sheath protoplasts (BSP) were isolated and purified fromfour C₄ species of the phosphoenolpyruvate (PEP) carboxykinasetype (Panicum maximum, P. texanum, Chloris gayana and Eriochloaborumensis), and cell organellses were separated from the BSPextract by differential centrifugation or sucrose density gradientcentrifugation. Separation of the organelles was ascertainedby the distribution of marker enzymes for chloroplasts, mitochondria,peroxisomes and cytoplasm. Contrary to the previous report [Rathnamand Edwards (1975) Arch. Biochem. Biophys. 171: 214], the distributionof PEP carboxykinase in BSP of P. maximum was the same as thatof UDP-glucose pyrophosphorylase, a marker for cytoplasm, andPEP carboxykinase activity was not recovered in the intact chloroplasts.The same results were obtained with P. texanum, C. gayana andE. borumensis. Therefore, we conclude that PEP carboxykinase is exclusivelylocalized in the cytoplasm of bundle sheath cells of C₄ plants. (Received July 23, 1983; Accepted October 17, 1983)     

Absorption of SO2 by Pecan (Carya illinoensis (Wang) K. Koch) and Alfalfa (Medicago sativa L.) and its Effect on Net Photosynthesis

Absorption rates of SO₂ by pecan (Carya illinoensis (Wang) K.Koch) leaflets exposed to 2.6, 5.2, and 7.8 mg SO₂ m^–3were measured over a 2 h period. SO₂ was rapidly absorbed bythe leaflets in all treatments during the initial 30–50min; the rate of uptake decreased to a rather constant levelthereafter. Total SO₂ absorbed during the 2 h period was 15.6,25.6, and 38.9 nmol cm^–2 for the low, medium, and highSO₂ concentrations, respectively. Reductions in net photosyntheticrates were proportional to ambient SO₂ concentrations and totalSO₂ absorbed. Partial photosynthetic recovery occurred in alltreatments during a 2 h post-treatment period and full recoveryoccurred during a 12 h dark period. Exposure to SO₂ resultedin slight increases in stomatal and boundary layer resistancesto CO₂ and substantial increases in residual resistances. Absorptionrates of SO₂ by alfalfa (Medicago saliva L.) exposed to 5.2mg SO₂m^–3 for 1 h were approximately double those of pecanexposed to the same ambient SO₂ concentration. Alfalfa net photosyntheticrates were reduced 74% after 1 h exposure to 5.2 mg SO₂ m^–3while a depression of 42% occurred in pecan.     

Ultrastructural Changes in Chloroplasts of Quercus rotundifolia Lam. in Response to Evernic Acid

The amount of total chlorophyll, chlorophylls a and b as wellas the ratio of a to b decreased in chloroplasts isolated fromQuercus rotundifolia leaves, kept for 17 d in a solution of35.5 µM evernic acid in 1 mM Na HCO₃, when compared withthe chloroplasts of control leaves (kept in NaHCO₃). The chloroplastsin the spongy parenchyma were smaller and the amount of starchand plastoglobuli lower. The number of grana per chloroplastsection, the number of thylakoids per grana and the height ofgrana stacks were also less in the chloroplasts of leaves treatedwith evernic acid. Quantitative ultrastructural differenceswere determined by means of electron microscopy and image analysistechniques. Quercus rotundifolia Lam., chloroplasts, ultrastructure, lichens, evernic acid     

Effects of Continuous SO2 Fumigation on SH-containing Compounds in Two Wheat Cultivars of Different Sensitivities

Two cultivars (Mec and Chiarano) of wheat (Triticum aestivum)were exposed to constant low levels of SO₂ (35, 75 and 120 nll^–1) over a period of 4 months. In previous studies Mechas been shown to be more sensitive to SO₂ and this has beenconfirmed in the present study where Mec showed a greater tendencythan Chiarano to accumulate soluble non-protein SH compounds,principally glutathione and cysteine. The reduced glutathione to oxidized glutathione ratio (GSH/GSSG)increased significantly in Mec with SO₂ treatment, but no changein the activity of glutathione reductase was observed. Key words: Wheat, SO₂ fumigation, SH-compounds     

Enzyme system catalyzing formation of cis-3-hexenal and n-hexanal from linolenic and linoleic acids in Japanese silver (Farfugium iaponicum Kitamura) leaves

Leaf alcohol (cis-3-hexenol) and leaf aldehyde (trans-2-hexenal)are responsible for the green odor in leaves and fruits. cis-3-Hexenal,a precursor of cis-3-hexenol and trans-2-hexenal, was producedfrom linolenic acid by a homogenate of Farfugium japonicum (Japanesesilver) leaves. n-Hexanal was produced from linoleic acid bya homogenate of the leaves. The enzyme system catalyzing formationof C₆-aldehydes from linolenic and linoleic acids was localizedin chloroplast lamellae, and required oxygen for reaction. C₁₈-unsaturatedfatty acids such as linolenic acid, linoleic acid and -linolenicacid, which have carboxyl groups and cis-1, cis-4-pentadienesystems including a double bond at C-12, acted as substrates,and C₆-aldehydes (cis-3-hexenal or n-hexanal), but not C₉-aldehydes,were produced from them. The properties of the enzyme systemin chloroplasts were as follows: optimal pH 7.0; stable at pH5 to 7; thermolabile and no activity at 50?C. These propertieswere very similar to those of tea chloroplasts. The enzyme systemcould be solubilized from chloroplasts by 2% Triton X-100, butwas very unstable in solubilized form. (Received July 9, 1976; )     

Fine Structure of Acid Mist Treated Sitka Spruce Needles: Open-top Chamber and Field Experiments

Sitka spruce grafts (clones DF and 141) grown in open-top chambers(OTC) and ‘mature ’, 6 –8m tall Sitka spruce(clone DF) grown in the field were exposed to acid mist containingan equimolar ion mixture of H₂SO₄and NH₄NO₃at pH2.5. Mist wasapplied 4 times a week (4 x1mm) in the OTC experiment and twicea week (2 x2mm) on average in the field experiment, betweenMay and Oct. 1991. Samples for light and electron microscopywere collected in Nov. and Jan. following acid mist treatment.Acid mist significantly decreased the amount of calcium depositedin the outer epidermal cell walls, the reduction being mostpronounced in the OTCs. Ultrastructurally, acid mist causeda significant increase in chloroplast and grana width. Othersymptoms associated with acid mist included swelling of chloroplastthylakoids, chloroplast protrusions, cytoplasm vacuolization,increase in large lipid accumulations and sickle-shaped chloroplastthylakoids. In the OTCs, acid mist hastened the acquisitionof frost hardening in both clones. In the field, the controltrees exhibited more frost injury than the acid mist treatedtrees suggesting, again, that acid mist had either hastenedor enhanced the stage of frost hardiness of treated trees. Ingeneral, acid mist induced changes were more pronounced in theOTCs than in the field. Picea sitchensis; Sitka spruce; acid mist; fine structure; calcium oxalate; frost hardening     

Control of ascorbic acid efflux in rat luteal cells: role of intracellular calcium and oxygen radicals

In luteal cells, prostaglandin (PG)F_2a mobilizes intracellular calcium concentration ([Ca]_i), generates reactive oxygen species (ROS), depletes ascorbic acid (AA) levels, inhibits steroidogenesis, and ultimately induces cell death. We investigated the hypothesis that [Ca]_i mobilization stimulates ROS, which results in depletion of cellular AA in rat luteal cells. We used a self-referencing AA-selective electrode that noninvasively measures AA flux at the extended boundary layer of single cells and fluorescence microscopy with fura 2 and dichlorofluorescein diacetate (DCF-DA) to measure [Ca]_i and ROS, respectively. Menadione, a generator of intracellular superoxide radical (), PGF_2a, and calcium ionophore were shown to increase [Ca]_i and stimulate intracellular ROS. With calcium ionophore and PGF_2a, but not menadione, the generation of ROS was dependent on extracellular calcium influx. In unstimulated cells there was a net efflux of AA of 121.5 ± 20.3 fmol · cm^–1 · s^–1 (mean ± SE, n = 8), but in the absence of extracellular calcium the efflux was significantly reduced (10.3 ± 4.9 fmol · cm^–1 · s^–1; n = 5, P < 0.05). PGF_2a and menadione stimulated AA efflux, but calcium ionophore had no significant effect. These data suggest two AA regulatory mechanisms: Under basal conditions, AA efflux is calcium dependent and may represent recycling and maintenance of an antioxidant AA gradient at the plasma membrane. Under luteolytic hormone and/or oxidative stress, AA efflux is stimulated that is independent of extracellular calcium influx or generation of ROS. Although site-specific mobilization of calcium pools and ROS cannot be ruled out, the release of AA by PGF_2a-stimulated luteal cells may occur through other signaling pathways. luteolysis; apoptosis; self-referencing microelectrode     

Anion-dependent changes of energy transfer in chloroplasts II. Relationships of the coupling factor to light-induced proton transport and absorbance change at 515 nm

Roles of the coupling factor in light-induced proton transportand 515-nm absorption change were investigated in chloroplastswashed with high concentrations of Tris salts (pH 7.2). Washingthe chloroplasts with Tris-HCl and Tris-HNO₃ buffers diminishedboth the light-induced pH rise and absorbance change at 515-nm,while Tris-H₂SO₄ buffer was much less effective. Inhibited activitiescould be restored by replacement of the coupling factor afterextraction with EDTA. N,N'-dicyclohexylcarbodiimide also restoredboth activities. Effects of various anions on the proton pumpand 515-nm shift were also investigated. The order of effectivenesswas NO₃^–>Cl^–>SO₄^2–. The role of thecoupling factor and its mode of action; the action mechanismsof Tris and anionsn energy transducing processes in chloroplasts,photophosphorylation, proton transport and absorbance changeat 515 nm, are discussed. ¹Present address: Biology Department, College of Science andEngineering, Ryukyu University, Naha, Okinawa, Japan. (Received June 27, 1972; )     

Changes of Anatomical Features, Photosynthesis and Ribulose Bisphosphate Carboxylase-Oxygenase Content of Mango Leaves

Changes in anatomical and physiological features, includingchanges in amount per unit area of anthocyanin and chlorophyll,in leaves of seedling mango (Mangifera indica L. cv. Irwin)trees were determined to understand what controls the rate ofphotosynthesis (Pn) at various stages of development. The youngleaves of seedling trees contained high concentrations of anthocyanin.During enlargement of leaves, the disappearance of anthocyaninand the accumulation of chlorophyll occurred concomitantly;the anthocyanin content began to decrease markedly once theleaf area had reached a maximum. During the early period ofleaf development, the thickness of mesophyll tissue decreasedtemporarily, but when the length of the leaf reached half thatof a mature leaf, the mesophyll began to thicken again. Smallstarch grains appeared in the chloroplasts of the young leavesand chloroplast nucleoids (ct-nuclei) were distributed throughoutthe chloroplasts. When leaves matured, ct-nuclei were displacedto the periphery of chloroplasts because of the accumulationof large starch grains. Compared with young leaves, green andmature leaves contained greater concentrations of ribulose bisphosphatecarboxylase-oxygenase (RuBisCO) protein. The results of immunocytochemicalexamination of RuBisCO under the light microscope reflectedthe results of electrophoresis measurements of RuBisCO. Pn waslow during the chocolate-coloured stage of early leaf development.In green and mature leaves Pn was higher; the average Pn was7·6 mg CO₂ dm^-2 h^-1 under light at intensities above500 µmol m^-2 s^-1.Copyright 1995, 1999 Academic Press Mangifera indica L., mango leaf, chloroplast nucleoids, chloroplast ultrastructure, starch accumulation, anthocyanin, chlorophyll, DAPI staining, SDS-PAGE, immunocytochemical technique