Photosynthesis and Other Traits in Relation to Chloroplast Number during Soybean Leaf Senescence

Soon after attaining full expansion, soybean (Glycine max [L.] Merr.) leaves enter a senescence phase marked by decline in photosynthetic rate and the progressive loss of chloroplast activity and composition. Our primary goal was to determine if this loss could be accounted for by sequential degradation of whole chloroplasts or by simultaneous degeneration of all chloroplasts. Total photosynthesis (TPs) measured as ¹⁴CO₂ uptake, chloroplast number, ribulose 1,5-bisphosphate carboxylase activity, uncoupled photosynthetic electron transport activity, soluble protein content, and chlorophyll content declined progressively during the 37 days after full leaf expansion. During this period, chloroplast number per unit leaf area was constant for all genotypes studied. We conclude that leaf senescence may be a two-stage process wherein the first stage chloroplast activity and composition declines, but chloroplast numbers do not change. During a brief terminal stage (11 days in our experiment), whole chloroplasts may be lost as well. As a second objective we wished to determine if variation in single-leaf total photosynthetic rate among soybean cultivars is related to corresponding variation in chloroplast number and/or chloroplast activity/composition. By comparing the means for three cultivars known to have rapid leaf TPs and for the three known to have slow TPs, we found the former group to be superior to the latter for all the previously mentioned leaf physiological traits. This superiority was related primarily to differences in chloroplast number and only secondarily to differences in activity and composition per chloroplast.     

Studies on genetic male-sterile soybeans : v. Effects of male-sterility on the function and glycerolipid composition of chloroplast thylakoids

Soybean (Glycine max L. Merr.) germplasm, isogenic except for loci controlling male sterility (ms₁), was utilized to study the effects of reproductive development on certain aspects of photosynthesis. Plants were sampled at various times between flowering (77 days after transplanting) and maturity (147 days after transplanting). During that period photosynthetic rates declined more rapidly in the male-sterile genotypes than male-fertile genotypes; and after 105 days, the sterile genotypes maintained low but relatively constant carbon exchange rates. The decline of leaf photosynthesis in the male-sterile genotype occurred concomitantly with an inhibition of the photosynthetic electron transport chain associated with photosystem II. Changes in photosystem I activities, cytochrome f levels, and chlorophyll a/b ratios per se were not responsible for the decline in whole leaf photosynthesis. These conditions were independent of the source of nitrogen nutrition. Lipid analyses of the thylakoids revealed that a loss of phosphatidylglycerol was highly correlated with the inhibition of photosystem II activity. These results suggested a relation between the decline in leaf carbon exchange and the decline in photosynthetic electron transport activity.     

Limiting Factors in Photosynthesis: II. IRON STRESS DIMINISHES PHOTOCHEMICAL CAPACITY BY REDUCING THE NUMBER OF PHOTOSYNTHETIC UNITS

It has been proposed that Fe stress may be used in the study of limiting factors in photosynthesis as an experimental means of varying photochemical capacity in vivo (Plant Physiol 1980 65: 114-120). In this paper the effect of Fe stress on photosynthetic unit number, size, and composition was investigated by measuring P₇₀₀, cytochrome (Cyt) f, chlorophyll (Chl) a, and Chl b in sugar beet leaves. The results show that when Fe stress reduced Chl per unit area by 80% (from 60 to 12 micrograms per square centimeter), it decreased the number of P₇₀₀ molecules per unit area by 88% and Cyt f per unit area by 86%; over the same range the Chl to P₇₀₀ ratio increased by 37% but there was no significant change in the Chl to Cyt f ratio. These data suggest that Fe stress decreases photochemical capacity and Chl per unit area by diminishing the number of photosynthetic units per unit leaf area.     

The Heterogeneity of Structural and Functional Photosynthetic Characteristics of Mesophyll Chloroplasts in Various Parts of Mature or Senescing Leaf Blade of Two Maize (Zea Mays L.) Genotypes

Differences in ultrastructural parameters of mesophyll cell (MC) chloroplasts, contents of photosynthetic pigments, and photochemical activities of isolated MC chloroplasts were studied in the basal, middle, and apical part of mature or senescing leaf blade of two maize genotypes. A distinct heterogeneity of leaf blade was observed both for structural and functional characteristics of chloroplasts. In both mature and senescing leaves the shape of MC chloroplasts changed from flat one in basal part of leaf to nearly spherical one in leaf apex. The volume density of granal thylakoids decreased from leaf base to apex in both types of leaves examined, while the amount of intergranal thylakoids increased in mature leaves but decreased in senescing leaves. The most striking heterogeneity was found for the quantity of plastoglobuli, which strongly increased with the increasing distance from leaf base. The differences in chloroplast ultrastructure were accompanied by differences in other photosynthetic characteristics. The Hill reaction activity and activity of photosystem 1 of isolated MC chloroplasts decreased from leaf base to apex in mature leaves. Apical part of senescing leaf blade was characterised by low contents of chlorophyll (Chl) a and Chl b, whereas in mature leaves, the content of Chls as well as the content of total carotenoids (Car) slightly increased from basal to apical leaf part. This was reflected also in the ratio Chl (a+b)/total Car; the ratio of Chl a/b did not significantly differ between individual parts of leaf blade. Both genotypes examined differed in the character of developmental gradient observed along whole length of leaf blade.     

Photosynthesis in leaves of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L. infected with tobacco mosaic virus

In tobacco leaves inoculated with tobacco mosaic virus (TMV), changes in chlorophyll (Chl) and carotenoid contents, parameters of slow Chl fluorescence kinetics, i.e. the maximum quantum yield of photosystem (PS2) photochemistry F_v/F_m, the effective quantum yield of photochemical energy conversion in PS2 Φ₂, ratio of quantum yields of photochemical and concurrent non-photochemical processes in PS2 F_v/F₀, non-photochemical quenching (NPQ), and photochemical activities of isolated chloroplasts from systemically infected tobacco leaves were investigated. We compared two successive stages of infection, the first in the stage of vein clearing at 9^th day post inoculation (dpi) and the second at 22^nd dpi when two different regions, i.e. light- (LGI) or dark-green (DGI) islands in the infected leaf were apparent and symptoms were fully developed. These two different regions were measured separately. The Chl and carotenoid contents in infected leaves decreased with a progression of infection and were lowest in LGI in the second stage. Also the ratio of Chl a/b declined in similar manner. The maximum quantum yield of PS2 photochemistry F_v/F_m, was decreased in the following order: first stage, DGI, and LGI. The same is true for the ratio F_v/F₀. The decrease of Φ₂ in infected leaves declined as compared to their controls. On the contrary, NPQ increased in infected leaves, the highest value was found in the first infection stage. Photochemical activities of the whole electron transport chain in isolated chloroplasts dramatically declined with the progression of symptoms, the lowest value was in LGI. Similarly, but to a lesser extent, the activity of PS2 in isolated chloroplasts decreased in infected leaves. Generally, the most marked impairment of the photosynthetic apparatus was manifested in the LGI of infected leaves.     

Chloroplast osmotic adjustment and water stress effects on photosynthesis

Previous studies have suggested that chloroplast stromal volume reduction may mediate the inhibition of photosynthesis under water stress. In this study, the effects of spinach (Spinacia oleracea, var `Winter Bloomsdale') plant water deficits on chloroplast photosynthetic capacity, solute concentrations in chloroplasts, and chloroplast volume were studied. In situ (gas exchange) and in vitro measurements indicated that chloroplast photosynthetic capacity was maintained during initial leaf water potential (Ψ_w) and relative water content (RWC) decline. During the latter part of the stress period, photosynthesis dropped precipitously. Chloroplast stromal volume apparently remained constant during the initial period of decline in RWC, but as leaf Ψ_w reached −1.2 megapascals, stromal volume began to decline. The apparent maintenance of stromal volume over the initial RWC decline during a stress cycle suggested that chloroplasts are capable of osmotic adjustment in response to leaf water deficits. This hypothesis was confirmed by measuring chloroplast solute levels, which increased during stress. The results of these experiments suggest that stromal volume reduction in situ may be associated with loss of photosynthetic capacity and that one mechanism of photosynthetic acclimation to low Ψ_w may involve stromal volume maintenance.     

Limiting Factors in Photosynthesis: III. Effects of Iron Nutrition on the Activities of Three Regulatory Enzymes of Photosynthetic Carbon Metabolism

When Fe was withheld from sugar beets (Beta vulgaris L. CV F58-44H1), the activities of NADP-glyceraldehyde-3-phosphate dehydrogenase, fructose 1,6-bisphosphatase, and ribulose 5-phosphate kinase were not diminished, while chlorophyll per area was decreased by 75%. On resupplying Fe, chlorophyll per area increased to control levels within 5 days, whereas the activities of the three enzymes remained approximately constant. These results support the view advanced earlier (Terry 1980 Plant Physiol 65: 114-120) that the photosynthetic effects of Fe deprivation are mediated by changes in the lamellar components of chloroplasts and not by changes in stromal enzymes involved in photosynthetic carbon reduction.     

Effects of drought stress on fluorescence characteristics of photosystem II in leaves of Plectranthus scutellarioides

Drought stress has multiple effects on the photosynthetic apparatus. Herein, we aimed to study the effect of drought stress on fluorescence characteristics of PSII in leaves of Plectranthus scutellarioides and explore potentially underlying mechanisms. Plants of P. scutellarioides were grown in a greenhouse and subjected to drought (DS, drought-stressed) or daily irrigation (control group). Leaf chlorophyll (Chl) index and induction kinetics curves of Chl a fluorescence and the JIP-test were used to evaluate effects of drought lasting for 20 d. Our results showed that both the leaf and soil relative water content decreased with increasing treatment duration. The leaf Chl index was reduced to half in the DS plants compared with the control group after 20 d. The minimal fluorescence in the DS plants was higher than that in the control plants after 10 d of the treatment. Maximum photochemical efficiency and lateral reactivity decreased with increasing treatment duration in the DS plants. With the continuing treatment, values of absorption flux per reaction center (RC), trapped energy flux per RC, dissipated energy flux per RC, and electron transport flux per RC increased in the earlier stage in the DS plants, while obviously decreased at the later stage of the treatment. In conclusion, drought stress inhibited the electron transport and reduced PSII photochemical activity in leaves of P. scutellarioides.     

Limiting Factors in Photosynthesis: VI. Regeneration of Ribulose 1,5-Bisphosphate Limits Photosynthesis at Low Photochemical Capacity

Earlier work (SE Taylor, N Terry [1984] Plant Physiol 75: 82-86) has shown that the rate of photosynthesis may be colimited by photosynthetic electron transport capacity, even at low intercellular CO₂ concentrations. Here we monitored leaf metabolites diurnally and the activities of key Calvin cycle enzymes in the leaves of three treatment groups of sugar beet (Beta vulgaris L.) plants representing three different in vivo photochemical capacities, i.e. Fe-sufficient (control) plants, moderately Fe-deficient, and severely Fe-deficient plants. The results show that the decrease in photosynthesis with Fe deficiency mediated reduction in photochemical capacity was through a reduction in ribulose 1,5-bisphosphate (RuBP) regeneration and not through a decrease in ribulose 1,5-bisphosphate carboxylase/oxygenase activity. Based on measurements of ATP and NADPH and triose phosphate/3-phosphoglycerate ratios in leaves, there was little evidence that photosynthesis and RuBP regeneration in Fe-deficient leaves were limited directly by the supply of ATP and NADPH. It appeared more likely that photochemical capacity influenced RuBP regeneration through modulation of enzymes in the photosynthetic carbon reduction cycle between fructose-6-phosphate and RuBP; in particular, the initial activity of ribulose-5-phosphate kinase was strongly diminished by Fe deficiency. Starch and sucrose levels changed independently of one another to some extent during the diurnal period (both increasing in the day and decreasing at night) but the average rates of starch or sucrose accumulation over the light period were each proportional to photochemical capacity and photosynthetic rate.     

Activation of NADP-Malate Dehydrogenase, Pyruvate,Pi Dikinase, and Fructose 1,6-Bisphosphatase in Relation to Photosynthetic Rate in Maize

The activity and extent of light activation of three photosynthetic enzymes, pyruvate,Pi dikinase, NADP-malate dehydrogenase (NADP-MDH), and fructose 1,6-bisphosphatase (FBPase), were examined in maize (Zea mays var Royal Crest) leaves relative to the rate of photosynthesis during induction and under varying light intensities. There was a strong light activation of NADP-MDH and pyruvate,Pi dikinase, and light also activated FBPase 2- to 4-fold. During the induction period for whole leaf photosynthesis at 30°C under high light, the time required to reach half-maximum activation for all three enzymes was only 1 minute or less. After 2.5 minutes of illumination the enzymes were fully activated, while the photosynthetic rate was only at half-maximum activity, indicating that factors other than enzyme activation limit photosynthesis during the induction period in C₄ plants.

Under steady state conditions, the light intensity required to reach half-maximum activation of the three enzymes was similar (300-400 microEinsteins per square meter per second), while the light intensity required for half-maximum rates of photosynthesis was about 550 microEinsteins per square meter per second. The light activated levels of NADP-MDH and FBPase were well in excess of the in vivo activities which would be required during photosynthesis, while maximum activities of pyruvate,Pi dikinase were generally just sufficient to accommodate photosynthesis, suggesting the latter may be a rate limiting enzyme.

There was a large (5-fold) light activation of FBPase in isolated bundle sheath strands of maize, whereas there was little light activation of the enzyme in isolated mesophyll protoplasts. In mesophyll protoplasts the enzyme was largely located in the cytoplasm, although there was a low amount of light-activated enzyme in the mesophyll chloroplasts. The results suggest the chloroplastic FBPase in maize is primarily located in the bundle sheath cells.

     

Effects of cucumber mosaic virus infection on electron transport and antioxidant system in chloroplasts and mitochondria of cucumber and tomato leaves

We examined the responses of the photosynthetic and respiratory electron transport and antioxidant systems in cell organelles of cucumber ( Cucumis sativus L.) and tomato ( Lycopersicon esculentum Mill.) leaves to infection of cucumber mosaic virus (CMV) by comparing the gas exchange, Chl fluorescence, respiratory electron transport, superoxide dismutase (SOD, EC 1.15.1.1) and ascorbate–glutathione (AsA–GSH) cycle enzymes and the production of H₂O₂ in chloroplasts, mitochondria and soluble fraction in virus-infected and non-infected leaves. Long-term CMV infection resulted in decreased photosynthesis and respiration rates. Photosynthetic electron flux to carbon reduction, respiratory electron transport via both complex I and complex II and also the Cyt respiration rate all significantly decreased, while photosynthetic alternative electron flux and alternative respiration significantly increased. These changes in electron transport were accompanied by a general increase in the activities of SOD/AsA–GSH cycle enzymes followed by an increased H₂O₂ accumulation in chloroplasts and mitochondria. These results demonstrated that disturbance of photosynthetic and respiratory electron transport by CMV also affected the antioxidative systems, thereby leading to oxidative stress in various organelles.     

Effects of drought stress on the evergreen Quercus ilex L., the deciduous Q. robur L. and their hybrid Q. × turneri Willd.

Five-year-old trees of deciduous Quercus robur L., evergreen Q. ilex L., and their semideciduous hybrid, Q. × turneri Willd. (var. pseudoturneri), growing in pots, were subjected to drought stress by withholding water for 18–22 days, until leaf water potentials decreased below ?2 MPa. Gas-exchange rates, oxygen evolution, and modulated chlorophyll (Chl) fluorescence measurements revealed that by strong stomata closure and declining photosynthetic capacity down to approximately 50%, all three taxa responded with strongly reduced photosynthesis rates. In Q. robur, photochemical quenching of the drought-stressed plants was much lower than in nonstressed controls. Dissection of the occurring events in the photosynthetic electron transport chain by fast Chl fluorescence induction analysis with the JIP-test were discussed.     

Image analysis of chlorophyll fluorescence transients for diagnosing the photosynthetic system of attached leaves

A new image instrumentation system for quantitative analysis of the rapid change in intensity of chlorophyll fluorescence during dark-light transition (CFI, chlorophyll fluorescence induction), which is a sensitive indicator of the various reactions of photosynthesis, was developed and its performance was evaluated. This system made it possible to resolve CFI at any small leaf area (about 1 square millimeter) of a whole leaf when the plant was illuminated by blue-green light at more than 50 micromoles photons per square meter per second. In order to test the usefulness of this system, we applied it to analyze the effect of SO₂ on photosynthetic apparatus in attached sunflower leaves. Dynamic CFI imaging over the whole single leaf, where there was no visible injury, indicated not only the local changes in photosynthetic activity but also the site of inhibition in photosynthetic electron transport system in chloroplasts. The new instrumentation system will be useful for the analytical diagnosis of various stress-actions on plants in situ.     

Influence of leaf age on photosynthesis, enzyme activity, and metabolite levels in wheat

The rate of photosynthesis under high light (1000 micromole quanta per square meter per second) and at 25°C was measured during development of the third leaf on wheat plants and compared with the activity of several photosynthetic enzymes and the level of metabolites. The rate of photosynthesis reached a maximum the 7th day after leaf emergence and declined thereafter. There was a high and significant correlation between the rate of photosynthesis per leaf area and the activities of the enzymes ribulose 5-phosphate kinase (r = 0.91), ribulose 1,5-bisphosphate (RuBP) carboxylase (r = 0.94), 3-phosphoglycerate (PGA) kinase (r = 0.82), and fructose 1,6-bisphosphatase (r = 0.80) per leaf area. There was not a significant correlation of photosynthesis rate with chlorophyll content. The rate of photosynthesis was strongly correlated with the level of PGA (r = 0.85) and inversely correlated with the level of triose phosphate (dihydroxyacetone phosphate and glyceraldehyde 3-phosphate) (r = 0.92). RuBP levels did not change much during leaf development; therefore photosynthesis rate was not correlated with the level of RuBP. The rate of photosynthesis was at a maximum when the ratio of PGA/triose phosphate was high, and when the ratio of RuBP/PGA was low. Although several enzymes change in parallel with leaf development, the metabolite changes suggest the greatest degree of control may be through RuBP carboxylase. The sucrose content of the leaf was highest under high rates of photosynthesis. There was no evidence that later in leaf development, photosynthesis (measured under high light and at 25°C) was limited by utilization of photosynthate.     

Effects of sulfur on the photosynthesis of intact leaves and isolated chloroplasts of sugar beets

Effects of sulfur on photosynthesis in sugar beets (Beta vulgaris L. cv. F58-554H1) were studied by inducing sulfur deficiency and determining changes in the photosynthesis of whole attached leaves and of isolated chloroplasts. The rates of photosynthetic CO₂ uptake by intact leaves, photoreduction of ferricyanide, cyclic and noncyclic photophosphorylation of isolated chloroplasts, and the rate of CO₂ assimilation by ribulose diphosphate carboxylase, decreased with decrease in total leaf sulfur from 2500 to about 500 μg g⁻¹ dry weight. Sulfur deficiency reduced photosynthesis through an effect on chlorophyll content, which decreased linearly with leaf sulfur, and by decreasing the rate of photosynthesis per unit chlorophyll. There was only a small effect of sulfur deficiency on stomatal diffusion resistance to CO₂ until leaf sulfur decreased below 1000 μg g⁻¹ when stomatal resistance became a more significant proportion of the total diffusion resistance to CO₂. Light respiration rates were positively correlated with photosynthesis rates and dark respiration was unchanged as leaf sulfur concentrations declined.     

Structural and functional organization of the photosynthetic apparatus in halophytes with different strategies of salt tolerance

The specific features of the structural and functional organisation of the photosynthetic apparatus (PSA) were studied in wild halophytes representing three strategies of salt tolerance: euhalophyte Salicornia perennans, crynohalophyte Limonium gmelinii, and glycohalophyte Artemisia santonica. The sodium content in aboveground parts of the plants corresponded to the strategy of salt tolerance. The photosynthetic cells of the euhalophyte were large and contained a higher number of chloroplasts than those in other species. In contrast, the number of cells per a leaf area unit was lower in S. perennans as compared to cryno- and glycohalophytes. Thereupon, the cell and chloroplast surface area per leaf area unit declined in the following sequence: A. santonica > L. gmelinii > S. perennans. However, the large cells of euhalophyte contained chloroplasts of larger sizes with 4- to 5-fold higher chlorophyll (Chl) content per chloroplast and Chl concentration in chloroplast volume unit. Also, chloroplasts of S. perennans were characterised by the higher content of glyco- and phospholipids. Qualitative composition of fatty acids (FA) in lipids isolated from the chloroplast-enriched fraction was similar in all three species; however, the index of unsaturation of FA was higher in glycohalophyte A. santonica than those in two other species. Under natural condition, PSA of all three halophytes showed high resistance to soil salinity. The results indicated tolerance of PSII to the photodamage in halophytes. The high rate of electron transport through PSII can be important to prevent oxidative damage of PSA in halophytes under strong light and hight temperature in vivo. Thus, the strategy of salt tolerance is provided by both the leaf anatomical structure and the ultrastructure of photosynthetic membranes, which is determined in particular by the specific composition of lipids.     

Diminution of photosynthesis in rice (Oryza sativa L.) seedlings under elevated CO2 concentration and increased temperature

The photosynthetic responses to elevated CO₂ concentration (EC) at ambient and ambient +4°C temperature were aßsessed in the second leaf of rice (Oryza sativa L.) seedlings. The duration of different leaf developmental phases, as characterised by changes in photosynthetic pigment contents and photochemical potential, was protracted in the seedlings grown under EC. On the other hand, a temporal shift in the phases of development with an early onset of senescence was observed in the seedlings grown under EC at ambient +4°C temperature. The contents of carotenoids, ß-carotene, and xanthophyll cycle pigments revealed that EC downregulated the protective mechanism of photosynthetic apparatus against oxidative damages, whereas this mechanism assumed higher significance under EC at ambient +4°C temperature. We observed an enhancement in electron transport activity, photochemical potential, and net photosynthesis in spite of a loss in photostasis of photosynthesis under EC. On the other hand, the loss in photostasis of photosynthesis was exacerbated under EC at ambient +4°C temperature due to the decline in electron transport activity, photochemical potential, and net photosynthesis.     

Changes in photosynthetic capacity and antioxidant enzymatic systems in micropropagated <Emphasis Type="Italic">Zingiber officinale</Emphasis> plantlets during their acclimation

Ginger (Zingiber officinale Rosc.) plantlets were propagated in vitro and acclimated under different photosynthetic photon flux densities (60 and 250 μmol m⁻² s⁻¹ = LI and HI, respectively). Increases in chlorophyll (Chl) content and Chl a/b ratio were found under both irradiances. In vitro plantlets (day 0) exhibited a low photosynthesis, but chloroplasts from in vitro leaves contained well developed grana and osmiophillic globules. Photoinhibition in leaves formed in vitro was characterized by decrease of photochemical efficiency and quantum efficiency of photosystem 2 photochemistry in HI treatment during acclimation. The new leaves formed during acclimation in both treatments showed a higher photosynthetic capacity than the leaves formed in vitro. Also activities of antioxidant enzymes of micropropagated ginger plantlets changed during acclimation.     

Photosynthetic Electron Transport,Photophosphorylation, and Antioxidants in Two Ecotypes of Reed (Phragmites Communis Trin.) from Different Habitats

We compared chloroplast photochemical properties and activities of some chloroplast-localised enzymes in two ecotypes of Phragmites communis, swamp reed (SR, C₃-like) and dune reed (DR, C₄-like) plants growing in the desert region of north-west China. Electron transport rates of whole electron transport chain and photosystem (PS) 2 were remarkably lower in DR chloroplasts. However, the electron transport rate for PS1 in DR chloroplasts was more than 90 % of the activity similar in the SR chloroplasts. Activities of Mg²⁺-ATPase and cyclic and non-cyclic photophosphorylations were higher in DR chloroplasts than in the SR ones. The activities of chloroplast superoxide dismutase (SOD) and ascorbate peroxidase (APX), both localised at or near the PS1 complex and serving to scavenge active oxygen around PS1, and the content of ascorbic acid, a special substrate of APX in chloroplast, were all higher in DR chloroplasts. Hence reed, a hydrophytic plant, when subjected to intense selection pressure in dune habitat, elevates its cyclic electron flow around PS1. In consequence, it provides extra ATP required by C₄ photosynthesis. Combined high activities of active oxygen scavenging components in DR chloroplasts might improve protection of photosynthetic apparatus, especially PS1, from the damage of reactive oxygen species. This offers new explanation of photosynthetic performance of plant adaptation to long-term natural drought habitat, which is different from those, subjected to the short-term stress treatment or even to the artificial field drought.     

Growth of Photoheterotrophic Cells of Peanut (Arachis hypogaea L.) in Still Nutrient Medium

Cell suspension cultures were established from the callus proliferation of leaf explants of 10- to 12-day-old seedlings of the peanut (Arachis hypogaea L. var. TMV-3). The cells could be cultivated in both agitated and still media, the latter promoting more of chlorophyll (Chl) synthesis. High Chl content (210-240 micrograms Chl per gram fresh weight), yield of free and pipetable cells, presence of all the pigments in the same ratio as that of the leaf tissue, and high rates of O₂ evolution (140-170 micromoles O₂ per milligram Chl per hour) were some of the desirable features of the still-grown cell cultures. However, considerable variations with regard to the above characters were observed between the cell cultures of different varieties of the peanut.

O₂ evolution by the cultured cells was dependent on exogenous supply of HCO₃⁻. A well-developed photosynthetic apparatus as evidenced from photosystem I and photosystem II activities of the isolated chloroplasts and variable fluorescence measurements with the cell cultures was further documented by electron microscopic evidence of distinct granal stackings in chloroplasts and sodium dodecyl sulfate-polyacrylamide gel separation of thylakoid membranes into P700 Chl a protein complex and light-harvesting Chl a/b complex. Evidence is presented for the relative increase in the Chl associated with P700 Chl a protein complex in contrast to the light-harvesting Chl a/b complex in the cultured cells as compared to intact leaf.