Localization of photosynthetic electron transport components in mesophyll and bundle sheath chloroplasts of Zea mays

The organization of the electron transport components in mesophyll and bundle sheath chloroplasts of Zea mays was investigated. Grana-containing mesophyll chloroplasts (chlorophyll a to chlorophyll b ratio of about 3.0) possessed the full complement of the various electron transport components, comparable to chloroplasts from C₃ plants. Agranal bundle sheath chloroplasts ($\text{Chl}\text{a}\text{Chl}\text{b}\text{> 5.0}$) contained the full complement of photosystem (PS) I and of cytochrome (cyt) f but lacked a major portion of PS II and its associated Chl $\text{a}\text{b}$ light-harvesting complex (LHC), and most of the cyt b₅₅₉. The kinetic analysis of system I photoactivity revealed that the functional photosynthetic unit size of PS I was unchanged and identical in mesophyll and bundle sheath chloroplasts. The results suggest that PS I is contained in stroma-exposed thylakoids and that it does not receive excitation energy from the Chl $\text{a}\text{b}$ LHC present in the grana. A stoichiometric parity between PS I and cyt f in mesophyll and bundle sheath chloroplasts indicates that biosynthetic and functional properties of cyt f and P₇₀₀ are closely coordinated. Thus, it is likely that both cyt f and P₇₀₀ are located in the membrane of the intergrana thylakoids only. The kinetic analysis of PS II photoactivity revealed the absence of PS II_αfrom the bundle sheath chloroplasts and helped identify the small complement of system II in bundle sheath chloroplasts as PS II_β. The distribution of the main electron transport components in grana and stroma thylakoids is presented in a model of the higher plant chloroplast membrane system.     

Photochemical properties of mesophyll and bundle sheath chloroplasts of maize

Several photochemical and spectral properties of maize (Zea mays) bundle sheath and mesophyll chloroplasts are reported that provide a better understanding of the photosynthetic apparatus of C₄ plants. The difference absorption spectrum at 298 K and the fluorescence excitation and emission spectra of chlorophyll at 298 K and 77 K provide new information on the different forms of chlorophyll a in bundle sheath and mesophyll chloroplasts: the former contain, relative to short wavelength chlorophyll a forms, more long wavelength chlorophyll a form (e.g. chlorophyll a 693 and chlorophyll a 705) and less chlorophyll b than the latter. The degree of polarization of chlorophyll a fluorescence is 6% in bundle sheath and 4% in mesophyll chloroplasts. This result is consistent with the presence of relatively high amounts of oriented long wavelength forms of chlorophyll a in bundle sheath compared to mesophyll chloroplasts. The relative yield of variable, with respect to constant, chorophyll a fluorescence in mesophyll chloroplasts is more than twice that in bundle sheath chloroplast. Furthermore, the relative yield of total chlorophyll a fluorescence is 40% lower in bundle sheath compared to that in mesophyll chloroplasts. This is in agreement with the presence of the higher ratio of the weakly fluorescent pigment system I to pigment system II in bundle sheath than in mesophyll chloroplast. The efficiency of energy transfer from chlorophyll b and carotenoids to chlorophyll a are calculated to be 100 and 50%, respectively, in both types of chloroplasts. Fluorescence quenching of atebrin, reflecting high energy state of chloroplasts, is 10 times higher in mesophyll chloroplasts than in bundle sheath chloroplasts during noncyclic electron flow but is equal during cyclic flow. The entire electron transport chain is shown to be present in both types of chloroplasts, as inferred from the antagonistic effect of red (650 nm) and far red (710 nm) lights on the absorbance changes at 559 nm and 553 nm, and the photoreduction of methyl viologen from H₂O. (The rate of methyl viologen photoreduction in bundle sheath chloroplasts was 40% of that of mesophyll chloroplasts.)     

Chlorophyll and peptide compositions in the two photosystems of marine green algae

The molar ratios of chlorophyll a to b in the thalli of marine green algae were between 1.5 and 2.2, being appreciably lower than the ratio between 2.8 and 3.4 found for the leaves of higher plants and the cells of fresh-water green algae. The ratio of chlorophylls to P-700 in these marine algae was also lower than that in higher plants. The $\text{a}\text{b}$ ratios in the pigment proteins of Photosystems 1 and 2 separated by polyacrylamide-gel electrophoresis from sodium dodecyl sulfate-solubilized chloroplasts of four species of marine green algae, Bryopsis maxima, Cheatomorpha spiralis, Enteromorpha compress and Ulva conglobata, were approximately 5 and 1, which are considerably smaller than the ratios, 7 and 2, respectively, found for the pigment proteins of the two photosystems of higher plants separated by the same technique. The chloroplasts of Bryopsis maxima and Cheatomorpha spiralis lacked two of the peptides associated with Photosystem II, which are present in the chloroplasts of Spinacia oleracea and Taraxacum officinale.     

Iron nutrition-mediated chloroplast development

Membrane development in chloroplasts was explored by resupplying iron to iron-deficient sugar beet (Beta vulgaris L. cv F58-554H1) and monitoring changes in lamellar components during regreening. The synthesis of chlorophyll a, chlorophyll b, and Q, the first stable electron acceptor of photosystem II, exhibited a lag phase during the first 24 to 48 hours of resupply. In contrast, the per area amounts of P₇₀₀ and cytochrome f increased linearly over the first 48 hours. During the early regreening period, the Q to P₇₀₀ ratio was 2.6 and decreased to 0.7 after 96 hours of regreening. The rate of photosynthesis (net CO₂ uptake) per chlorophyll increased during the first 48 hours of resupply, then by 96 hours decreased to values typical of control plants. The results suggest that there was preferential synthesis of the measured photosystem I components during the first 24 to 48 hours, while from 48 to 96 hours there was rapid synthesis of all components. The iron nutrition-mediated chloroplast development system provides a useful experimental approach for studying biomembrane synthesis and structural-functional relations of the photosynthetic apparatus.     

Spectral, Physical, and Electron Transport Activities in the Photosynthetic Apparatus of Mesophyll Cells and Bundle Sheath Cells of Digitaria sanguinalis (L.) Scop

Isolated mesophyll cells and bundle sheath cells of Digitaria sanguinalis were used to study the light-absorbing pigments and electron transport reactions of a plant which possesses the C₄-dicarboxylic acid cycle of photosynthesis. Absorption spectra and chlorophyll determinations are presented showing that mesophyll cells have a chlorophyll a-b ratio of about 3.0 and bundle sheath cells have a chlorophyll a-b ratio of about 4.5. The absorption spectrum of bundle sheath cells has a greater absorption in the 700 nm region at liquid nitrogen temperature, and there is a relatively greater amount of a pigment absorbing at 670 nm in the bundle sheath cells compared to the mesophyll cells. Fluorescence emission spectra, at liquid nitrogen temperature, of mesophyll cells have a fluorescence 730 nm-685 nm ratio of about 0.82 and bundle sheath cells have a ratio of about 2.84. The reversible light-induced absorption change in the region of P₇₀₀ absorption is similar in both cell types but bundle sheath cells exhibit about twice as much total P₇₀₀ change as mesophyll cells on a total chlorophyll basis. The delayed light emission of bundle sheath cells is about one-half that of mesophyll cells. Both mesophyll cells and bundle sheath cells evolve oxygen in the presence of Hill oxidants with the mesophyll cells exhibiting about twice the activity of bundle sheath cells, and both activities are inhibited by 1 μM 3-(3,4-dichlorophenyl)-1, 1-dimethylurea. Ferredoxin nicotinamide adenine dinucleotide phosphate reductase is present in both cells although it is about 3- or 4-fold higher in mesophyll cells than in bundle sheath cells. Glyceraldehyde 3-P dehydrogenases, both nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate, are equally distributed in the two cell types on a chlorophyll basis. Malic enzyme is localized in the bundle sheath cells.     

Photosystem electron-transport capacity and light-harvesting antenna size in maize chloroplasts

Spectrophotometric and kinetic measurements were applied to yield photosystem (PS) stoichiometries and the functional antenna size of PSI, PSII_α, and PSII_β in Zea mays chloroplasts in situ. Concentrations of PSII and PSI reaction centers were determined from the amplitude of the light-induced absorbance change at 320 and 700 nm, which reflect the photoreduction of the primary electron acceptor Q of PSII and the photooxidation of the reaction center P700 of PSI, respectively. Determination of the functional chlorophyll antenna size (N) for each photosystem was obtained from the measurement of the rate of light absorption by the respective reaction center. Under the experimental conditions employed, the rate of light absorption by each reaction center was directly proportional to the number of light-harvesting chlorophyll molecules associated with the respective photosystem. We determined N_P700 = 195, N_α = 230, N_β = 50 for the number of chlorophyll molecules in the light-harvesting antenna of PSI, PSII_α, and PSII_β, respectively. The above values were used to estimate the PSII/PSI electron-transport capacity ratio (C) in maize chloroplasts. In mesophyll chloroplasts C > 1.4, indicating that, under green actinic excitation when Chl a and Chl b molecules absorb nearly equal amounts of excitation, PSII has a capacity to turn over electrons faster than PSI. In bundle sheath chloroplasts C < 1, suggesting that such chloroplasts are not optimally poised for linear electron transport and reductant generation.     

The P700-chlorophyll a-protein. Isolation and some characteristics of the complex in higher plants

A P700-chlorophyll a-protein complex has been purified from several higher plants by hydroxylapatite chromatography of Triton X-100-dissociated chloroplast membranes. The isolated material exhibits a red wavelength maximum at 677 nm, major spectral forms of chlorophyll a at 662, 669, 677, and 686 nm, a chlorophyll/P700 ratio of $\text{40–45}\text{1}$, and contains only chlorophyll a and β-carotene of the photosynthetic pigments present in the chloroplast. The spectral characteristics and composition of the higher plant material are homologous to those of the P700-chlorophyll a-protein previously isolated from blue-green algae; however, unlike the blue-green algal component, cytochromes f and b₆ are associated with the higher plant material. Evidence is presented that a chlorophyll a-protein termed “Complex I” which can be isolated from sodium dodecyl sulfate extracts of chloroplast membranes is a spectrally altered form of the eucaryotic P700-chlorophyll a-protein. The isolation procedure described in this paper is a more rapid technique for obtaining the heart of photosystem I than presently exists; furthermore, the P700 photooxidation and reduction kinetics in the fraction are improved over those in other isolated components showing the same enrichment of P700. It appears very probable that the heart of photosystem I is organized in the same manner in all chlorophyll a-containing organisms.     

Photochemical activities and organization of photosynthetic apparatus of C3 and C4 plants grown under different light intensities

Changes in the photochemical activities, influenced by variation in the growth light intensity, were followed in typical C₃ (Phaseolus, Ipomoea) and C₄ (Amaranthus, Sorghum) plants. Progressive decrease in the growth light intensity accelerated the O-P fluorescence induction in whole leaves. Such acceleration of the fluorescence kinetics was found to be not due to enhanced photosystem II activity but possibly a result of reduced rate of electron flow between the two photosystems. This is supported by 4 lines of evidence: (1) by the Hill activity determined in the presence of electron acceptors functioning before and after plastoquinone; (2) the photosynthetic unit size determined after flash excitation showing variations that were apparently too small to account for the changes observed fluorescence induction; (3) modification of the kinetics of secondrange light-induced absorbance changes at 520 nm; and (4) absence of significant changes in the ratio of P₇₀₀/total chlorophyll ratio. The P₇₀₀/cytochrome f ratio, however, increased from the usual 1–1.5 to 3–4 in plants grown under 9% sunlight. Increase in the P₇₀₀/cytochrome f ratio was found to be due to a decrease in the cytochrome f/chlorophyll ratio, and this was due to perhaps to a simultaneous increase in chlorophyll and decrease in cytochrome content.     

Photochemical activities and organization of photosynthetic apparatus of C3 and C4 plants grown under different light intensities

Changes in the photochemical activities, influenced by variation in the growth light intensity, were followed in typical C₃ (Phaseolus, Ipomoea) and C₄ (Amaranthus, Sorghum) plants. Progressive decrease in the growth light intensity accelerated the O-P fluorescence induction in whole leaves. Such acceleration of the fluorescence kinetics was found to be not due to enhanced photosystem II activity but possibly a result of reduced rate of electron flow between the two photosystems. This is supported by 4 lines of evidence: (1) by the Hill activity determined in the presence of electron acceptors functioning before and after plastoquinone; (2) the photosynthetic unit size determined after flash excitation showing variations that were apparently too small to account for the changes observed fluorescence induction; (3) modification of the kinetics of second-range light-induced absorbance changes at 520 nm; and (4) absence of significant changes in the ratio of P₇₀₀/total chlorophyll ratio. The P₇₀₀/cytochrome f ratio, however, increased from the usual 1–1.5 to 3–4 in plants grown under 9% sunlight. Increase in the P₇₀₀/cytochrome f ratio was found to be due to a decrease in the cytochrome f/chlorophyll ratio, and this was due to perhaps to a simultaneous increase in chlorophyll and decrease in cytochrome content.     

Chlorophyll composition of Photosystems IIα, IIβ and I in tobacco chloroplasts

The antenna composition of the Photosystems II_α, II_β and I was studied in tobacco chloroplasts. Absorbance spectra, recorded at 4 K, were analyzed for the wild type and the mutants Su/su and Su/su var. Aurea, containing higher concentrations of the photosystems. With chloroplasts of Su/su we measured the action spectra of the three photosystems from 625 to 690 nm. Above 675 nm absorption by Photosystem I dominated. This sytem had a maximum at 678 nm and a shoulder at 660 nm. Of the long-wavelength chlorophyll a forms, absorbing at 690, 697 and 705 nm at 4 K, which are generally assigned to Photosystem I, the 697 nm form occurred in an amount of four molecules per reaction center of Photosystem I in each type of chloroplast. The Photosystem II_α spectrum was characterized by maxima at 650 and 672 nm, showing clearly the participation of the chlorophyll a and b containing light-harvesting complex. In the mutants the light-harvesting complex has a chlorophyll a to chlorophyll b ratio of more than 1; the amount of the 672 nm chlorophyll a was normal, whereas the amount of chlorophyll b was markedly decreased in the mutants relative to the wild type. The Photosystem II_β spectrum mainly consisted of a band at 683 nm.     

THE CHLOROPHYLL-PROTEIN COMPOUND OF THE GREEN LEAF

1. Aqueous extracts of spinach and Aspidistra leaves yield highly opalescent preparations which are not in true solution. Such extracts differ markedly from colloidal chlorophyll in their spectrum and fluorescence. The differences between the green leaf pigment and chlorophyll in organic solvents are shown to be due to combination of chlorophyll with protein in the leaf. 2. The effect of some agents on extracts of the chlorophyll-protein compound has been investigated. Both strong acid and alkali modify the absorption spectrum, acid converting the compound to the phaeophytin derivative and alkali saponifying the esterified groups of chlorophyll. Even weakly acid solutions (pH 4.5) denature the protein. Heating denatures the protein and modifies the absorption spectrum and fluorescence as earlier described for the intact leaf. The protein is denatured by drying. Low concentrations of alcohol or acetone precipitate and denature the protein; higher concentrations cause dissociation liberating the pigments. 3. Detergents such as digitonin, bile salts, and sodium desoxycholate clarify the leaf extracts but denature the protein changing the spectrum and other properties. 4. Inhibiting agents of photosynthesis are without effect on the absorption spectrum of the chlorophyll-protein compound. 5. The red absorption band of chlorophyll possesses the same extinction value in organic solvents such as ether or petroleum ether, and in aqueous leaf extracts clarified by digitonin although the band positions are different. Using previously determined values of the extinction coefficients of purified chlorophylls a and b, the chlorophyll content of the leaf extracts may be estimated spectrophotometrically. 6. It was found that the average chlorophyll content of the purified chloroplasts was 7.86 per cent. The protein content was 46.5 per cent yielding an average value of 16.1 parts per 100 parts of protein. This corresponds to a chlorophyll content of three molecules of chlorophyll a and one of chlorophyll bfor the Svedberg unit of 17,500. It is suggested that this may represent a definite combining ratio of a and b in the protein molecule.     

The state of detergent solubilised light-harvesting chlorophyll-a/b protein complex as monitored by picosecond time-resolved fluorescence and circular dichroism

Steady-state and picosecond time-resolved fluorescence techniques in conjunction with circular dichroism have been used to study the light-harvesting chlorophyll-a/b protein complex (LHC) isolated from pea chloroplasts. In particular, the effect of changing the detergent / chlorophyll ratio on the state of the LHC has been investigated. Our results have been interpreted in light of the known protein geometry of the LHC in 2-dimensional crystals (Kühlbrandt, W. (1984) Nature 307, 478–479). The fluorescence lifetime data reveals 1 / e-lifetimes of 3.53 (±0.04) ns and 1.10 (±0.01) ns for a stable, efficiently energy-transferring state of the LHC. Subnanosecond lifetimes are observed under conditions leading to aggregation, while a long component of 5.50 (±0.16) ns corresponding to free Chl a is found when the detergent / chlorophyll ratio is high. The circular dichroism shows a major Chl-b exciton, a Chl-a / b exciton and a further ‘quenching’ Chl-b exciton. These have been attributed to: a C₃ symmetric Chl-b interaction for which the intact C₃ protein trimer geometry is a prerequisite; a dimeric Chl-a / b interaction, the presence of which is critically dependent on the detergent type; and a further Chl-b interaction which arises from the presence of aggregated trimers, respectively. We have found that the degree of heterogeneity with respect to the oligomeric state of the pigment-protein trimers is dependent upon the detergent / chlorophyll ratio used. Low detergent / chlorophyll ratios result in extensive aggregation of the trimers with a geometry similar to that found in 2-dimensional crystals of the LHC. Moderate detergent conditions yield predominantly non-aggregated trimers. Excess detergent conditions result in considerable chromophore heterogeneity and loss of the main Chl-b exciton consistent with protein denaturation through an initial break up of the trimer geometry. From these results we believe that in vitro the minimum stable functional unit corresponds to a C₃ symmetric protein trimer.     

High photosynthetic rate of a chlorophyll mutant of cotton

In a chlorophyll mutant (virescent) and wild-type cotton (Gossypium hirsutum L.), a number of photosynthetic parameters have been measured and compared with those published for other chlorophyll mutants. (a) The photosynthetic rates at 230 w/m² (400-700 nm) from a tungsten lamp were 36.8 mg CO₂ fixed/dm²·hr (virescent) and 39.5 mg CO₂ fixed/dm²·hr (wild-type). On a chlorphyll basis, the photosynthetic rates were 36.8 and 12.1 mg CO₂ fixed/mg chl·hr, respectively. (b) The photosynthetic rates at 13 w/m² (400-700 nm) from a tungsten source were 7.1 mg CO₂ fixed/dm²·hr (virescent) and 7.4 mg CO₂ fixed/dm²·hr (wild-type). On a chlorophyll basis, the photosynthetic rates were 6.0 and 1.4 mg CO₂ fixed/mg chl·hr, respectively. (c) The chlorophyll a/b ratios of the virescent and wild-type leaves were 3.3 and 4.1 (d) The chlorophyll/carotenoid ratios for the virescent and wild-type leaves were 3.2 and 7.3, respectively. (e) The photosynthetic carbon metabolism of the chlorophyll mutant was through the reductive pentose phosphate cycle. (f) The CO₂ compensation points for the virescent and wild-type plants were similar. (g) The mutant and wild-type leaves have the same quantum yield in the red part of the visible spectrum, but the virescent leaves have a lower quantum yield in the blue part of the spectrum. (h) Virescent and wild-type leaves contain similar levels on a protein basis of several reductive pentose phosphate cycle enzymes.     

Nitrogen deposition limits photosynthetic response to elevated CO2 differentially in a dioecious species

Sexual dimorphisms of dioecious plants are important in controlling and maintaining sex ratios under changing climate environments. Yet, little is known about sex-specific responses to elevated CO₂ with soil nitrogen (N) deposition. To investigate sex-related physiological and biochemical responses to elevated CO₂ with N deposition, Populus cathayana Rehd. was employed as a model species. The cuttings were subjected to two CO₂ regimes (350 and 700???mol?mol^?1) with two N levels (0 and 5?g?N?m^?2?year^?1). Our results showed that elevated CO₂ and N deposition separately increased the total number of leaves, leaf area (LA), leaf mass, net photosynthetic rate (P _n), light saturated photosynthetic rate (P _max), chlorophyll a (Chl a), and chlorophyll a to chlorophyll b ratio (Chl a/b) in both males and females of P. cathayana. However, the effects on LA, leaf mass, P _n, P _max, Chl a and Chl a/b were weakened under the combined treatment of elevated CO₂ and N deposition. Males had higher leaf mass, P _n, P _max, apparent quantum yield (??), carboxylation efficiency (CE), Chl a, Chl a/b, leaf N, and root carbon to N ratio (C/N) than did females under elevated CO₂ with N deposition. In contrast to males, females had significantly higher levels of soluble sugars in leaves and greater starch accumulation in roots and stems under the same condition. The results of the present work imply that P. cathayana females are more responsive and suffer from greater negative effects on growth and photosynthetic capacity than do males when grown under elevated CO₂ with soil N deposition.     

Inhibition of photosynthetic electron transport and formation of inactive chlorophyll in winter stressed Pinus silvestris

Kinetics of fluorescence at room temperature, electron transport and photooxidation of P700 and cytochrome f have been studied in chloroplasts isolated from active and winter stressed Pinus silvestris. The winter stress induced block in the electron transport chain between the two photosystems is close to the site of plastoquinone, since winter stress and DCMU caused the same type of inhibition of the reoxidation of the primary electron acceptor Q of photosystem II. No winter inhibition of the electron transport between cytochrome f and P700 was observed. Time course studies of P700 photooxidation in chloroplasts of active and winter stressed pine have shown that the photosynthetic unit size must be about equal in the two types of chloroplasts. An apparent increase of the photosynthetic unit size was induced by winter stress, as revealed by the high chlorophyll/P700 ratio of winter stressed pine. The phenomenon is explained by the formation of photosynthetically inactive chlorophyll. Low-temperature fluorescence emission spectra were recorded when either chlorophyll a (433 nm) or chlorophyll b (477 nm) were preferentially excited. Winter stress induced the formation of a chlorophyll a fraction emitting at 673 nm. This chlorophyll is most likely derived from the chlorophyll a antennae of the two photosystems, and it probably contributes to the photosynthetically inactive pool of chlorophyll in winter stressed pine. The light harvesting chlorophyll a/b complex is relatively resistant to winter stress.     

Spectral characterization of five chlorophyll-protein complexes

Sodium dodecyl sulfate-solubilized chloroplast internal membranes of higher plants (cowpea [Vigna unguiculata L. Walp], chinese cabbage [Brassica chinensi L.], and tobacco [Nicotiana tabacum L.]) are resolved by polyacrylamide gel electrophoresis into two chlorophyll a- and three chlorophyll a,b-proteins. A small portion (about 15%) of the membrane chlorophyll migrates as a component of high electrophoretic mobility and presumably consists of detergent-complexed, protein-free pigment.

One of the chlorophyll a-proteins is qualitatively similar to the P₇₀₀ chlorophyll a-protein but contains a much larger proportion of total chlorophyll (about 30%) than previously reported. The second chlorophyll a-protein is a recently discovered component of the membrane and accounts for about 7% of the total chlorophyll. The absorption and fluorescence emission spectra of these two chlorophyll a-proteins differ.

The three chlorophyll a,b-proteins are components of the chloroplast membrane chlorophyll a,b-light-harvesting complex which was previously resolved as a single chlorophyll-protein band. The two additional chlorophyll a,b-proteins observed in our work probably represent larger aggregates contained within that membrane complex which are preserved under the solubilization and electrophoretic conditions used here.

     

Photosynthetic CO(2) Fixation Products and Activities of Enzymes Related to Photosynthesis in Bermudagrass and Other Plants

After a 5-second exposure of illuminated bermudagrass (Cynodon dactylon L. var. `Coastal') leaves to ¹⁴CO₂, 84% of the incorporated ¹⁴C was recovered as aspartate and malate. After transfer from ¹⁴CO₂-air to ¹²CO₂-air under continuous illumination, total radioactivity decreased in aspartate, increased in 3-phosphoglyceric acid and alanine, and remained relatively constant in malate. Carbon atom 1 of alanine was labeled predominantly, which was interpreted to indicate that alanine was derived from 3-phosphoglyceric acid. The activity of phosphoenolpyruvate carboxylase, alkaline pyrophosphatase, adenylate kinase, pyruvate-phosphate dikinase, and malic enzyme in bermudagrass leaf extracts was distinctly higher than those in fescue (Festuca arundinacea Schreb.), a reductive pentose phosphate cycle plant. Assays of malic enzyme activity indicated that the decarboxylation of malate was favored. Both malic enzyme and NADP⁺-specific malic dehydrogenase activity were low in bermudagrass compared to sugarcane (Saccharum officinarum L.). The activities of NAD⁺-specific malic dehydrogenase and acidic pyrophosphatase in leaf extracts were similar among the plant species examined, irrespective of the predominant cycle of photosynthesis. Ribulose-1, 5-diphosphate carboxylase in C₄-dicarboxylic acid cycle plant leaf extracts was about 60%, on a chlorophyll basis, of that in reductive pentose phosphate cycle plants.     

Antenna replacement in the evolutionary origin of chloroplasts

The endosymbiotic origin of chloroplasts from unicellular cyanobacteria is presently beyond doubt. Oxygenic photosynthesis is based on coordinated action of the two photosystems (PS), PS I and PS II, cooperating with several variants of the pigment antenna. In cyanobacteria, red algae, and glaucophytes, phycobilisomes (PBS) act as antennae, while in terrestrial plants, as well as in most macro- and microalgae, antennae are formed by chlorophyll a/b- and chlorophyll a/c-containing proteins. Advantages and disadvantages of the PBS antenna compared to other light-harvesting complexes form the basis for adaptive variations of the antenna in the course of development of eukaryotic photosynthesis. During the evolution of the “green” and “chromophyte” lineages of the chloroplasts, PBS, in spite of their optimal features of light absorption, were replaced by chlorophyll a/b- and chlorophyll a/c-containing light-harvesting complexes. Development of the cell wall associated with the limitation of motility and tissue formation in photosynthetic eukaryotes were the factors responsible for the antenna shift. The subsequent redistribution of cell resources in favor of cellulose biosynthesis required for increased CO₂ consumption, higher PS II levels, and greater number and density of the thylakoids in the chloroplasts, was incompatible with the energy-consuming and overly large PBS antenna.     

Effect of Long Term Exposure to High CO2 Concentrations on Photosynthetic Characteristics of Prunus Avium L. Plants

The effect of two elevated carbon dioxide concentrations, 700 µmol(CO₂) mol^–1 (C700) and 1 400 µmol(CO₂) mol^–1 (C1400), on photosynthetic performances of 1-year-old Prunus avium L. plant was studied. Plants grown at C700 were characterised by increased net photosynthetic rate (P _N) as compared to those grown at C1400. Plant photosynthetic adjustment to C1400 resulted in 27 % higher P _N than in control at atmospheric CO₂ concentration (C _a) at the beginning of the experiment (3–4 weeks) with a consequent decline to the end of the experiment. Thus, 1 400 µmol(CO₂) mol^–1 had short-term stimulatory effect on plant P _N. Both chlorophyll (Chl) a and b concentrations dramatically decreased during exposure to C1400. Compensation irradiance was increased by 57 % in C700 and by 87 % in C1400. Photochemical efficiency () was affected by balloon environment, however, a clear stimulatory effect of C700 was detected. Opposite influence of both elevated CO₂ concentrations on P _Nmax was established: slight increase by C700 (2.7 % at C_a), but considerable decrease by C1400 (63 % at C_a). Exposure to C700 enhanced compensation irradiance by 42 %, while C1400 by only 21 %. Either C700 or C1400 did not reduce stomatal conductance (g _s). Leaf area per plant (LAR) was more stimulated by C700 than by C1400. High unit area leaf mass, specific leaf area, and dry matter accumulation in roots without affecting tissue density characterised plants grown in C1400. However, when considering the root : shoot ratio, these plants allocated less carbon to the roots than plants from other treatments.     

Isolation and Characterization of a Light-Harvesting Chlorophyll a/b Protein Complex Associated with Photosystem I

A chlorophyll a/b protein complex has been isolated from a resolved native photosystem I complex by mildly dissociating sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The chlorophyll a/b protein contains a single polypeptide of molecular weight 20 kilodaltons, and has a chlorophyll a/b ratio of 3.5 to 4.0. The visible absorbance spectrum of the chlorophyll a/b protein complex showed a maximum at 667 nanometers in the red region and a 77 K fluorescence emission maximum at 681 nanometers. Alternatively, by treatment of the native photosystem I complex with lithium dodecyl sulfate and Triton, the chlorophyll a/b protein complex could be isolated by chromatography on Sephadex G-75. Immunological assays using antibodies to the P₇₀₀-chlorophyll a-protein and the photosystem II light-harvesting chlorophyll a/b protein show no cross-reaction between the photosystem I chlorophyll a/b protein and the other two chlorophyll-containing protein complexes.