[Show abstract][Hide abstract] ABSTRACT: Peas were grown in controlled environments (12h white fluorescent light. ∼47 μmol photons m-2 s 1/12 dark, 25 °C), using (1) 15-min far-red illumination at the end of each photoperiod (brief FR) to simulate the increase in the far-red/red ratio near the end of the day, and (2) high levels of supplementary far-red light (red:far-red ratio=0.04) during the entire photoperiod (long-term FR) to simulate extreme shade conditions under a plant canopy. Brief FR illumination led to marked morphological effects attributable to phytochrome regulation, namely, an increase in internodal length, but a decrease in leaflet area, chloroplast size and chlorophyll content per chloroplast compared with the control. Significantly, brief FR illumination had little or no effect on the amounts of the major chloroplast components (ribulose 1.5-biphosphate carboxylase, adenosine triphosphate synthase, cytochrome b/f complex and Photosystem II) relative to chlorophyll or Photosystem I, and the leaf photosynthetic capacities per unit chlorophyll were similar. In contrast, supplementing high levels of far-red light during the entire photoperiod not only led to the phytochrome effects above, but there was also a marked increase in leaf photosynthetic capacity per unit chlorophyll. due to increased amounts of the major chloroplast components relative to chlorophyll or Photosystem I. We hypothesize that supplementary far-red light, absorbed by Photosystem I, induced an increase in the major chloroplast components by a photosynthetic feedback mechanism. In fully greened leaves, we propose that the two photosystems themselves, rather than phytochrome, may be the predominent sensors of light quantity in triggering modulations of the stoichiometries of chloroplast components, which in turn lead to varying photosynthetic capacities.
[Show abstract][Hide abstract] ABSTRACT: The photosynthetic acclimation of Alocasia macrorrhiza (L.) G. Don, a species naturally occurring in deep shade in rainforests, has been studied in relation to a wide range of controlled irradiances during growth (~3-780 µmol photons m-2 s-1 of fluorescent or incandescent light, 10 h light/ 14 h dark). At the maximum growth irradiances, the light- and CO2-saturated rates of O2 evolution per unit leaf area were ~4 times as high as at low irradiance, and approached those of glasshouse-grown spinach. Growth at maximum irradiances reduced the quantum yield of O2 evolution only slightly.
Changes in the anatomy of leaf tissue, the ultrastructure of chloroplasts and the composition of chloroplast components accompanied the changes in photosynthetic functional characteristics. At low growth irradiance, palisade cell chloroplasts were preferentially located adjacent to the distal periclinal cell walls and had large granal stacks, and the destacked thylakoids had a very low surface charge density. In contrast, at higher growth irradiance, palisade cell chloroplasts were preferentially located adjacent to the anticlinal cell walls; they had small granal stacks, large stromal space, and a high surface charge density on the destacked thylakoids. The number of chloroplasts per unit section length increased with growth irradiance. Ribulosebisphosphate carboxylase activity per unit leaf area increased markedly with irradiance. Photosystem II, cytochrome f and latent ATPase activity per unit chlorophyll increased to a lesser extent. While the chlorophyll a/chlorophyll b ratio increased substantially with growth irradiance, the chlorophyll content per unit leaf area declined slightly.
Our results show that coordinated changes in the structure of leaf tissue, and the organisation and composition of chloroplast components are responsible for Alocasia being capable of acclimation to high as well as low irradiance.
[Show abstract][Hide abstract] ABSTRACT: The photosynthetic apparatus of plants responds to changing light quantity and quality with coordinated changes in both the light-harvesting antennae of the photosystems and the amounts of electron transport components and ATP synthase. These compositional modulations are accompanied by changes in thylakoid membrane organisation and photosynthetic capacity. It is now clear that there is a dynamic continuum of organisation and function of the photosynthetic apparatus from the appressed granal and non-appressed stroma thylakoids within a chloroplast, to different chloroplasts within a leaf, to leaves within and between species. While it is very unlikely that there is a unique solution to photosynthesis in the sun or shade, substantial changes in composition, and hence thylakoid membrane organisation and function, are elicited as part of sun/shade responses.
[Show abstract][Hide abstract] ABSTRACT: Immunoblotting was used to probe the reactivity of rabbit polyclonal antibodies against PS1I and PSI light-harvesting chlorophyll a/b-proteins of spinach (Spinacea oleracea L.) with the light-harvesting complexes of a siphonaceous marine alga, Codium, that have more chlorophyll b, siphonaxanthin and siphonein instead of the lutein. The spinach LHCII antibodies cross-reacted only with the apoproteins of Cod-ium LHCII. Antisera against the spinach LHCI apoproteins showed strong affinity for the apoproteins of Codium LHCI, and also reacted with the polypeptides of spinach LHCII and Codium LHCII. Our results indicate some similarities in the amino acid sequences between the Codium siphonaxanthin-Chl a/fe-proteins of LHCII and LHCI and the corresponding spinach lutein-chlorophyll a/b-proteins.
[Show abstract][Hide abstract] ABSTRACT: To probe the location of the carboxyl-terminus of the 28 kDa apoprotein of the light-harvesting chlorophyll a/b-protein complex of PS II (LHCII), an antibody was generated against a synthetic octapeptide corresponding to the C-terminal region of LHCII. The high specificity of the octapeptide antiserum was deonstrated by immunoblots and immunogold labelling. The octapeptide antiserum agglutinated destacked thylakoid membranes, but no significant agglutination occurred with inside-out vesicles suggesting that the COOH-terminus is located at the outer, stroma-exposed surface where the NH2-terminus is also located [(1983) J. Biol. Chem. 258, 9941-9948]. Our results support a model for LHCII with four transmembrane-spanning domains.
[Show abstract][Hide abstract] ABSTRACT: Some marine nematodes from anoxic mangrove mud-flats contain densely packed intracellular inclusions within their tissues which are visible by light microscopy. Transmission electron microscopy of sections of Sabatieria wieseri, Terschellingia longicaudata and Sphaerolaimus papillatus shows these inclusions within the intestinal and other cells. Energy dispersive analysis of secondary X-rays by scanning electron microscopy of thick epoxy sections shows concentrations of Si, P, S, K, Ca and Fe in their tissues. EDS of secondary X-ray emissions from thin epoxy unstained sections by transmission electron microscopy shows these elements together with Na, Zn and Al concentrated in the intracellular inclusions. The physiological implications of these observations are discussed.
[Show abstract][Hide abstract] ABSTRACT: Mixed cultures of Cellulomonas sp. and Azospirillum brasilense were grown with straw or cellulose as the carbon source under conditions favoring the fixation of atmospheric nitrogen. Rapid increases in cell numbers, up to 10 cells per g of substrate, were evident after 4 and 5 days of incubation at 30 degrees C for cellulose and straw, respectively. Nitrogen fixation (detected by acetylene reduction measured on parallel cultures) commenced after 2 and 4 days of incubation for straw and cellulose, respectively, and continued for the duration of the experiment. Pure cultures of Cellulomonas sp. showed an increase in cell numbers, but CO(2) production was low, and acetylene reduction was not detected on either cellulose or straw. Pure cultures of A. brasilense on cellulose showed an initial increase in cell numbers (10 cells per g of substrate) over 4 days, followed by a decline presumably caused by the exhaustion of available carbon substrate. On straw, A. brasilense increased to 10 cells per g of substrate over 5 days and then declined slowly; this growth was accompanied by acetylene reduction. Scanning electron micrographs of straw incubated with a mixed culture under the above conditions for 8 days showed cells of both species in close proximity to each other. Evidence was furnished that the close spatial relationship of cells from the two species facilitated the mutually beneficial association between them and thus increased the efficiency with which the products of straw breakdown were used for nitrogen fixation.
Applied and Environmental Microbiology 05/1986; 51(4):849-54. · 3.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Antibodies directed against purified cytochrome f, isolated from the cytochrome b/f complex of spinach chloroplasts, were used in on-grid immunogold labelling studies of spinach leaf tissue. Our results show unambiguously that cytochrome f, and hence the cytochrome b/f complex, is located in both appressed and non-appressed thylakoid membranes.
Cell Biology International Reports 09/1985; 9(8):715-21. DOI:10.1016/0309-1651(85)90079-7
[Show abstract][Hide abstract] ABSTRACT: The effect of light quality on the composition, function and structure of the thylakoid membranes, as well as on the photosynthetic rates of intact fronds from Asplenium australasicum, a shade plant, grown in blue, white, or red light of equal intensity (50 microeinsteins per square meter per second) was investigated. When compared with those isolated from plants grown in white and blue light, thylakoids from plants grown in red light have higher chlorophyll a/chlorophyll b ratios and lower amounts of light-harvesting chlorophyll a/b-protein complexes than those grown in blue light. On a chlorophyll basis, there were higher levels of PSII reaction centers, cytochrome f and coupling factor activity in thylakoids from red light-grown ferns, but lower levels of PSI reaction centers and plastoquinone. The red light-grown ferns had a higher PSII/PSI reaction center ratio of 4.1 compared to 2.1 in blue light-grown ferns, and a larger apparent PSI unit size and a lower PSII unit size. The CO(2) assimilation rates in fronds from red light-grown ferns were lower on a unit area or fresh weight basis, but higher on a chlorophyll basis, reflecting the higher levels of electron carriers and electron transport in the thylakoids.The structure of thylakoids isolated from plants grown under the three light treatments was similar, with no significant differences in the number of thylakoids per granal stack or the ratio of appressed membrane length/nonappressed membrane length. The large freeze-fracture particles had the same size in the red-, blue-, and white-grown ferns, but there were some differences in their density. Light quality is an important factor in the regulation of the composition and function of thylakoid membranes, but the effects depend upon the plant species.
[Show abstract][Hide abstract] ABSTRACT: A computational method for estimating the radial distribution function of particles seen in electron micrographs of freeze-fractured membranes is presented. The method overcomes the previous difficulty of measurements of this quantity from freeze-fracture electron micrographs--the systematic error due to the finite size of the sample areas in the freeze-fracture. By multiplying the measurements by weights specific to the shape of the sample area, the method lessens the errors due to the boundaries of the area and allows a synthesis of the average radial distribution function from counts over many small separate areas.
Computers in Biology and Medicine 02/1982; 12(4):319-22. DOI:10.1016/0010-4825(82)90035-X · 1.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The major intrinsic protein from spinach chloroplast membranes, the light-harvesting chlorophyll a/b-protein complex, contains two distinct polypeptides of Mr 23,500 and 26,000 and 31% lipid by weight, comprising five diacyl lipids and seven chlorophylls, together with some carotenoids, per 26,000-Mr polypeptide. The chlorophyll a/b ratio is 1.1. Low-temperature fluorescence emission spectra of the light-harvesting complex revealed a major peak at 681 nm with a shoulder of variable intensity at 695 nm. The 695-nm emission has been correlated with a progressive aggregation of the complex into two-dimensional, semi-crystalline sheets. To determine the role of the light-harvesting complex in cation-dependent thylakoid stacking, the purified complex has been quantitatively incorporated into liposomes containing the four major chloroplast diacyl lipids using a simple freeze-thaw technique. The proteoliposomes appeared largely as unilamellar vesicles, with diameters between 0.1 and 0.8 micron. Freeze-fracture analysis showed intramembrane particles of 8-10 nm corresponding to the incorporated complex. Both monovalent and divalent cations caused an immediate aggregation of the proteoliposomes, which was reversed at low cation concentrations and was largely inhibited by prior trypsin treatment. Since lipid vesicles themselves showed none of these effects, it is concluded that surface-exposed polypeptide regions of the light-harvesting complex are directly involved in thylakoid stacking in vivo.
European Journal of Biochemistry 07/1980; 107(2):345-54. DOI:10.1111/j.1432-1033.1980.tb06035.x · 3.58 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Chlorella fusca, strain 211-15, cells degreened in a nitrogen-deficient mineral growth medium in the light for 4-6 weeks were regreened for up to 24 hrs in a nitrogen rich medium that leads to synchronous cell division at 24-26 hrs. Structural changes in the plastid membranes during the regreening period were observed by thin section and freeze-fracture electron microscopy. Nitrogen-deficient plastids were found to have non-appressed lamellae, prolamellar body-like membrane aggregations, and only 2 types of freeze-fracture face. At this time no photosynthetic oxygen evolution could be demonstrated. After 6 hrs regreening the plastid lamellae had fused to form bands of appressed lamellae and the four types of freeze-fracture face, described previously, were visible. At this time photosynthetic oxygen evolution could be demonstrated. After 24 hrs regreening the plastids had an appearance typical of normally grown Chlorella and had commenced to divide. Supporting evidence for these developmental stages is presented from isolated chloroplast particle fractions. An unusual type of cell wall proliferation was observed in the nitrogen-deficient Chlorella cells that resulted in the laying down of several walls, each with a trilaminar component.
Archives of Microbiology 06/1975; 103(3):259-70. · 1.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Aggregations of approximately 11 nm spaced spherical particles are described in both freeze-fractured and chemically fixed bean and spinach leaf plastids. It is suggested that the aggregations were induced as a result of the dehydrating action of the cryoprotectants used. It is tentatively concluded, on the basis of previous reports, that the aggregations consist of Fraction I protein.
[Show abstract][Hide abstract] ABSTRACT: Chlorella fusca, strain 211-15, cells degreened in a nitrogen-deficient mineral growth medium in the light for 4–6 weeks were regreened for up to 24 hrs in a nitrogen rich medium that leads to synchronous cell division at 24–26 hrs. Structural changes in the plastid membranes during the regreening period were observed by thin section and freeze-fracture electron microscopy. Nitrogen-deficient plastids were found to have non-appressed lamellae, prolamellar body-like membrane aggregations, and only 2 types of freeze-fracture face. At this time no photosynthetic oxygen evolution could be demonstrated. After 6 hrs regreening the plastid lamellae had fused to form bands of appressed lamellae and the four types of freeze-fracture face, described previously, were visible. At this time photosynthetic oxygen evolution could be demonstrated. After 24 hrs regreening the plastids had an appearance typical of normally grown Chlorella and had commenced to divide. Supporting evidence for these developmental stages is presented from isolated chloroplast particle fractions.An unusual type of cell wall proliferation was observed in the nitrogen-deficient Chlorella cells that resulted in the laying down of several walls, each with a trilaminar component.
Archives of Microbiology 12/1974; 103(1):259-270. DOI:10.1007/BF00436359 · 1.67 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Chloroplasts were isolated from leaves of three species of tropical rainforest plants, Alocasia macrorrhiza, Cordyline rubra and Lomandra longifolia; these species are representative of extreme “shade” plants. It was found that shade plant chloroplasts contained 4–5 times more chlorophyll than spinach chloroplasts. Their chlorophyll a/chlorophyll b ratio was 2.3 compared with 2.8 for spinach. Electron micrographs of leaf sections showed that the shade plant chloroplasts contained very large grana stacks. The total length of partitions relative to the total length of stroma lamellae was much higher in Alocasia than in spinach chloroplasts. Freeze-etching of isolated chloroplasts revealed both the small and large particles found in spinach chloroplasts.Despite their increased chlorophyll content, low chlorophyll a/chlorophyll b ratio, and large grana, the shade plant chloroplasts were fragmented with digitonin to yield small fragments (D-144) highly enriched in Photosystem I, and large fragments (D-10) enriched in Photosystem II. The degree of fragmentation of the shade plant chloroplasts was remarkably similar to that of spinach chloroplasts, except that the subchloroplast fragments from the shade plants had lower chlorophyll a/chlorophyll b ratios than the corresponding fragments from spinach. The D-10 fragments from the shade plants had chlorophyll a/chlorophyll b ratios of 1.78-2.00 and the D-144 fragments ratios of 3.54–4.07. We conclude that Photosystems I and II of the shade plants have lower proportions of chlorophyll a to chlorophyll b than the corresponding photosystems of spinach. The lower chlorophyll a/chlorophyll b ratio of shade plant chloroplasts is not due to a significant increase in the ratio of Photosystem II to Photosystem I in these chloroplasts.The extent of grana formation in higher plant chloroplasts appears to be related to the total chlorophyll content of the chloroplast. Grana formation may simply be an means of achieving a higher density of light-harvesting assemblies and hence a more efficient collection of light quanta.
[Show abstract][Hide abstract] ABSTRACT: The mechanism of digitonin action on spinach chloroplasts was investigated by thin sectioning. Evidence is presented which shows that digitonin continues to modify membranes for many minutes after the addition of the fixative glutaraldehyde. However, the action of digitonin can be stopped by simultaneous fixation and dilution of the detergent. Such experiments indicate that the initial action of digitonin is to release stroma lamellae which in turn yield a Photosystem 1 fraction. This interpretation is further supported by a significant correlation between the ratio and the ratio of stroma to grana lamellae in spinach chloroplasts.
[Show abstract][Hide abstract] ABSTRACT: Class II spinach chloroplasts were fragmented by passage through the French pressure cell (French press), and the fragments were separated by fractional centrifugation. Fragments sedimenting between 1000 × g and 10000 × g (10K) have a lower chlorophylla/chloropyllb ratio and lower P-700 content than whole chloroplasts. Fragments sedimenting between 40000 × g and 160000 × g (160K) have a much higher chlorophylla/chlorophyllb ratio (6.0) and a much higher P-700 content (1 P-700 per 105 chlorophylls) than whole chloroplasts. The chlorophyll and cytochrome contents of the French press fractions are similar to those found in fractions isolated after digitonin disruption.The 160K fraction performs Photosystem 1 but not Photosystem 2 reactions. The 10K fraction contains both photosystems. Electrophoresis of sodium dodecyl sulfate solubilized 10K and 160K fractions gives further evidence for this distribution of photosystems.Thin sectioning and freeze fracturing show that the 160K fraction originates from stroma lamellae and the end membranes of grana stacks and contains only 110Åparticles. The 10K fraction originates from the partition regions of grana stacks and contains both 110 and 175Åparticles. This distribution of particles on fracture faces of stroma versus grana lamellae is shown to exist in freeze fractured Class I chloroplasts.These data demonstrate that both digitonin and French press treatments of chloroplasts initially break stroma lamellae and end membranes to yield small vesicles which contain only Photosystem 1.
[Show abstract][Hide abstract] ABSTRACT: A chlorophyll-deficient mutant of pea (Pisum sativum) was found as a spontaneous mutation of the variety Greenfeast. Total chlorophyll of the mutant leaves was about one-half that of normal pea leaves per mg dry weight, and the ratio of chl a:chl b ranged from 10 to 18, compared with 3 for normal pea. In each generation the mutant plants gave rise to normal and mutant plants and lethal plants with yellow leaves.For a normal pea plant, CO(2) uptake was saturated at about 60,000 lux, whereas with mutant leaves, the rate of CO(2) uptake was still increasing at 113,000 lux. At 113,000 lux the mutant and normal leaves showed similar rates of CO(2) fixation per unit area of leaf surface, but on a chlorophyll basis the mutant leaves were twice as active. Hill reaction measurements on isolated chloroplasts also showed that the mutant chloroplasts were saturated at higher intensities than the normal, and that the activity of the mutant was at least double that of the normal on a chlorophyll basis.It is suggested that the photosynthetic units of the mutant chloroplasts contain about half the number of chlorophyll molecules as compared to the normal photosynthetic units.Electron microscopy of leaf sections of normal and mutant leaves showed that the mutant chloroplasts contain fewer lamellae per chloroplast and fewer lamellae per granum. The lethal chloroplasts, which were virtually devoid of chlorophyll, were characterized by an absence of grana.