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1.
P700 enriched fractions were isolated from two brown algae and one diatom using sucrose density centrifugation after digitinin solubilization. They had a Chl a/P700 ratio of about 250 to 375 according to the species, they were enriched in long-wavelength absorbing Chl a and exhibited a fluorescence emission maximum at 77 K near 720 nm. They all presented a major polypeptide component at 66±2 kDa, but their polypeptide composition was rather complex and somewhat different from one species to another. Further solubilization with dodecylmaltoside of those native PSI particles allowed the separation of two or three fractions. The lightest, xanthophyll-rich, fraction was identified to be a light-harvesting complex. It contained no P700 and had a major polypeptide of molecular weight near 20 kDa (at the same molecular weight than the respective LH native fraction of each species) and exhibited a 77 K peak fluorescence emission at 685 nm. The other fractions were enriched in P700 and almost entirely depleted in xanthophylls. When two of them are present, they both exhibited a major polypeptide at 66±2 kDa and were totally devoid of the LH polypeptide, but the two fractions widely differed one from another in the abundance and molecular weight of the other polypeptide components. The most purified of these two fractions presented a composition similar to PSI core complex from green plants.Abbreviations LH light-harvesting - LHCII light-harvesting complex II of green plants - P700 reaction center chlorophyll of PSI  相似文献   

2.
Two types of chlorophyll-protein complexes of photosystem I (PSIa, PSIc) have been isolated from the membranes of Spirulina platensis using a Triton X-100 treatment and chromatography on DEAE-Toyopearl. The complexes are equally enriched with P700 (Chl: P700 = 100-110) but show different electrophoretic molecular masses--140 (PSIa) and 320 kDa (PSIc)--and differ in the content of long-wavelength absorbing Chl. PSIa has a typical PSI fluorescence band at 730 nm (F730) as the main band at 77 K, whereas PSIc is responsible for F760, the intensity of which depends on the redox state of P700. PSIc only shows 77 K light-induced variable fluorescence at 760 typical of Spirulina membranes and cells.  相似文献   

3.
Eight chlorophyll b deficient nuclear mutants of pea (Pisum sativum L.) have been characterized by low temperature fluorescence emission spectra of their leaves and by the ultrastructure, photochemical activities and polypeptide compositions of the thylakoid membranes. The room temperature fluorescence induction kinetics of leaves and isolated thylakoids have also been recorded. In addition, the effects of Mg2+ on the fluorescence kinetics of the membranes have been investigated. The mutants are all deficient in the major polypeptide of the light-harvesting chlorophyll a/b protein of photosystem II. The low temperature fluorescence emission spectra of aurea-5106, xantha-5371 and –5820 show little or no fluorescence around 730 nm (photosystem I fluorescence), but possess maxima at 685 and 695 nm (photosystem II fluorescence). These three mutants have low photosystem II activities, but significant photosystem I activities. The long-wavelength fluorescence maximum is reduced for three other mutants. The Mg2+ effect on the variable component of the room temperature fluorescence (685 nm) induction kinetics is reduced in all mutants, and completely absent in aurea-5106 and xantha-5820. The thylakoid membranes of these 2 mutants are appressed pairwise in 2-disc grana of large diameter. Chlorotica-1-206A and–130A have significant long-wavelength maxima in the fluorescence spectra and show the largest Mg2+ enhancement of the variable part of the fluorescence kinetics. These two mutants have rather normally structured chloroplast membranes, though the stroma regions are reduced. The four remaining mutants are in several respects of an intermediate type.Abbreviations Chl chlorophyll - CPI Chi-protein complex I, Fo, Fv - Fm parameters of room temperature chlorophyll fluorescence induction kinetics - F685, F695 and F-1 components of low temperature chlorophyll emission with maximum at 685, 695 and ca 735 nm, respectively - PSI photosystem I - PSII photosystem II - LHCI and LHCII light-harvesting chlorophyll a/b complexes associated with PSI and PSII, respectively - SDS sodium dodecyl sulfate  相似文献   

4.
The phylogenetic distribution of photosystem I-associated polypeptides was assessed by immunoblotting algal thylakoid membrane polypeptides with antisera generated against the P700-chlorophyll a protein (CC I) and a photosystem I light-harvesting chlorophyll-protein (LHC Ib). Polypeptides cross-reacting with the CC I apoprotein were found in 20 species representing four classes of unicellular algae. Polypeptides sharing antigenicity with spinach LHC Ib were observed only in algal species containing chlorophyll b. Tetraselmis spp. (Pleurastrophyceae), rich in chlorophyll b (Chl a:b 1.2), exhibited marked heterogeneity in the composition of their CC I and LHC Ib cross-reactive polypeptides. When immunoblotted with antisera against CC I, all Tetraselmis clones examined exhibited a 25-kD polypeptide in greater abundance than the 58-kD CC I apoprotein characteristic of higher plants and other green algal thylakoids. Three Tetraselmis clones (RG 6, RG 11, and RG 12) exhibited an 81-kD polypeptide with strong antigenicity toward the LHC Ib antisera, in contrast to the 17- to 24-kD cross-reactive polypeptides found in spinach, green algae, and one Tetraselmis clone (RG 5). Associated with the unique photosystem I polypeptide composition in Tetraselmis spp., Chl: P700 ratios for the group are 2–5 times greater than those observed for higher plants or other green algae. The chlorophyll b enrichment, unusual composition of photosystem I cross-reactive polypeptides, and heterogeneity of these polypeptides within isolates of Tetraselmis might make this genus useful for investigations of the functional organization of chlorophyll b in light-harvesting systems. These features also support the view of an alternative phyletic origin for the Pleurastrophyceae.  相似文献   

5.
Lin  Zhi-Fang  Peng  Chang-Lian  Lin  Gui-Zhu  Zhang  Jing-Liu 《Photosynthetica》2003,41(4):589-595
Two new yellow rice chlorophyll (Chl) b-less (lack) mutants VG28-1 and VG30-5 differ from the other known Chl b-less mutants with larger amounts of soluble protein and ribulose-1,5-bisphosphate carboxylase/oxygenase small sub-unit and smaller amounts of Chl a. We investigated the altered features of Chl-protein complexes and excitation energy distribution in these two mutants, as compared with wild type (WT) rice cv. Zhonghua 11 by using native mild green gel electrophoresis and SDS-PAGE, and 77 K Chl fluorescence in the presence of Mg2+. WT rice revealed five pigment-protein bands and fourteen polypeptides in thylakoid membranes. Two Chl b-less mutants showed only CPI and CPa pigment bands, and contained no 25 and 26 kDa polypeptides, reduced amounts of the 21 kDa polypeptide, but increased quantities of 32, 33, 56, 66, and 19 kDa polypeptides. The enhanced absorption of CPI and CPa and the higher Chl fluorescence emission ratio of F685/F720 were also observed in these mutants. This suggested that the reduction or loss of the antenna LHC1 and LHC2 was compensated by an increment in core component and the capacity to harvest photon energy of photosystem (PS) 1 and PS2, as well as in the fraction of excitation energy distributed to PS2 in the two mutants. 77 K Chl fluorescence spectra of thylakoid membranes showed that the PS1 fluorescence emission was shifted from 730 nm in WT rice to 720 nm in the mutants. The regulation of Mg2+ to excitation energy distribution between the two photosystems was complicated. 10 mM Mg2+ did not affect noticeably the F685/F730 emission ratio of WT thylakoid membranes, but increased the ratio of F685/F720 in the two mutants due to a reduced emission at 685 nm as compared to that at 720 nm.  相似文献   

6.
Analyses of chlorophylls a and b and P700 in the wheat leaves grown for 8 days under illumination with white light at different intensities suggested selective formation of photosystem 1 of the photosynthesis at low light intensities. This was confirmed for the two types of chloroplasts isolated from leaves grown at light intensities of 1.1 and 240 μ W/cm2, respectively, by measuring their pigment compositions, activities of photosystems 1 and 2, and absorption and fluorescence spectra. The chloroplasts developed at the low intensity showed properties only of photosystem 1 while those developed at the high intensity showed properties of both photosystems 1 and 2. Only photosystem 1 particles were obtained by fractionation of low intensity chloroplasts by treatment with digitonin followed by centrifugation, while high intensity chloroplasts could be fractionated into photosystem-1 and photosystem-2 particles. When the leaves grown at low light intensity were illuminated with strong light, photosystem 2 was developed. The fluorescence emission spectrum of low intensity chloroplasts at 77°K showed two peaks at 685 and 734 nm, and the spectrum of high intensity chloroplasts showed three peaks at 685, 697 and 740 nm.  相似文献   

7.
Acclimation to periodic high‐light stress was studied in tree seedlings from a neotropical forest. Seedlings of several pioneer and late‐succession species were cultivated under simulated tree‐fall gap conditions; they were placed under frames covered with shade cloth with apertures of different widths that permitted defined periods of daily leaf exposure to direct sunlight. During direct sun exposure, all plants exhibited a marked reversible decline in potential photosystem II (PSII) efficiency, determined by means of the ratio of variable to maximum Chl a fluorescence (Fv/Fm). The decline in Fv/Fm under full sunlight was much stronger in late‐succession than in pioneer species. For each gap size, all species exhibited a similar degree of de‐epoxidation of violaxanthin in direct sunlight and similar pool sizes of xanthophyll cycle pigments. Pool sizes increased with increasing gap size. Pioneer plants possessed high levels of β‐carotene that also increased with gap size, whereas α‐carotene decreased. In contrast to late‐succession plants, pioneer plants were capable of adjusting their Chl a/b ratio to a high value in wide gaps. The content of extractable UV‐B‐absorbing compounds was highest in the plants acclimated to large gaps and did not depend on the successional status of the plants. The results demonstrate a better performance of pioneer species under high‐light conditions as compared with late‐succession plants, manifested by reduced photoinhibition of PSII in pioneer species. This was not related to increased pool size and turnover of xanthophyll cycle pigments, nor to higher contents of UV‐B‐absorbing substances. High β‐carotene levels and increased Chl a/b ratios, i.e. reduced size of the Chl a and b binding antennae, may contribute to photoprotection in pioneer species.  相似文献   

8.
The light dependent chloroplast development of dark grown seedlings of Pinus silvestris L. was followed by analyses of chlorophyll content, chlorophyll a/b ratios, chlorophyll/P700 ratios, chlorophyll-protein complexes and structural changes. Low-temperature fluorescence emission spectra of isolated chloroplasts and separation of sodium dodecyl sulphate solubilized chlorophyll-protein complexes by gel electrophoresis showed that the chlorophyll-protein complexes of photosystem 1 (P700-CPa), photosystem II (PS II-CPa) and the light-harvesting complex LH–CPa/b were present in dark grown seedlings. The low-temperature fuoorescence emission maxima of isolated P700–CPa and PS II–CPa shifted towards longer wavelengths during greening in light, indicating a light induced change of the chlorophyll organisation in the two photosystems. Illumination caused LH–CPa/b to increase relative to P700–CPa, whereas the ratio between LH–CPa/b and PS II–CPa remained essentially constant. Analyses of low-temperature fluorescence spectra with or without 0.01 M Mg2+ showed that the Mg2+ controlled distribution of excitation energy into PS I was activated upon illumination of the seedlings. The photosynthetic unit size, as defined by the chlorophyll/P700 ratio, did not change over a 96 h illumination period, although the chlorophyll content increased about 6–fold during that time. This result and the constant electron transport rate per unit chlorophyll and time during chlorophyll accumulation provided evidence for a sequential development of the photosynthetic units when illuminating dark grown pine cotyledons. Electron micrographs showed that exposure of dark grown seedlings to light for 2 h caused the prolamellar body to disappear and grana to form. These changes occurred prior to substantial accumulation of chlorophyll or change in the ratio between LH–CPa/b and P700–CPa. However, both the water-splitting system of photosystem II and the Mg2+ controlled redistribution of excitation energy was activated during this period.  相似文献   

9.
A rather simple method of isolation of photosystem 2 fragments, which are highly purified from Photosystem 1 admixture, has been developed on the basis of combined action of detergents and differential centrifugation. The isolated fragments are characterized by insignificant content of P700 (one molecule per 10500 molecules of chlorophyll) and by high ratio of band values at 685 and 735 nm in the low temperature emission spectrum of fluorescence (F685/F735=5.9). The data on photochemical activity and ability for photoinduced changes in fluorescence prove that the activity of Photosystem 2 is retained both at the level of reaction centre operation and at that of water photooxidation with oxygen evolution.  相似文献   

10.
10% of the chlorophyll associated with a ‘native’ Photosystem (PS) I complex (110 chlorophylls/P-700) is chlorophyll (Chl) b. The Chl b is associated with a specific PS I antenna complex which we designate as LHC-I (i.e., a light-harvesting complex serving PS I). When the native PS I complex is degraded to the core complex by LHC-I extraction, there is a parallel loss of Chl b, fluorescence at 735 nm, together with 647 and 686 nm circular dichroism spectral properties, as well as a group of polypeptides of 24-19 kDa. In this paper we present a method by which the LHC-I complex can be dissociated from the native PS I. The isolated LHC-I contains significant amounts of Chl b (Chl ab ? 3.7). The long-wavelength fluorescence at 730 nm and circular dichroism signal at 686 nm observed in native PS I are maintained in this isolated complex. This isolated fraction also contains the low molecular weight polypeptides lost in the preparation of PS I core complex. We conclude that we have isolated the PS I antenna in an intact state and discuss its in vivo function.  相似文献   

11.
When grown heterotrophically in the dark on enriched culture medium, the pigment-deficient strain of Scenedesmus obliquus, mutant C-6E, is uniquely characterized by a complete deficiency in carotenoids and chlorophyll b while retaining a low level of chlorophyll a which is exclusively utilized in photosystem I-type reactions. The strain lacks photosystem II activity but exhibits all PS-I reactions tested, including P700 redox reactions, photoreduction of CO2 with hydrogen as electron donor, and O2 uptake following methyl viologen reduction. The mutant contains 10 times more P700 per chlorophyll than the wild type and develops the pigment-protein complex of PS-I, CP-I. The action spectrum for methyl viologen reduction compares favorable to the low temperature absorption spectrum of whole cells. Both the chlorophyll fluorescence excitation and emission spectra of pigment-protein complexes derived from cells of C-6E show patterns typical of PS-I. The strain lacks the LHCs and CP-II as well as their respective apoproteins. The absence of carotenoids appears to prevent the development of the normal variety of pigment-protein complexes and the accumulation of Chl b. This inability is also expressed by the presence of only single stranded thylakoid membranes in the chloroplast of C-6E. When heterotrophically grown cells of this mutant are exposed to white light of 8 or 22 W m?2, 50% of its chlorophyll is lost by photooxidation within 4 or 1.5 hours, respectively.  相似文献   

12.
Thermal emission and photochemical energy storage were examined in photosystem I reaction center/core antenna complexes (about 40 Chl a/P700) using photoacoustic spectroscopy. Satisfactory signals could only be obtained from samples bound to hydroxyapatite and all samples had a low signal-to-noise ratio compared to either PS I or PS II in thylakoid membranes. The energy storage signal was saturated at low intensity (half saturation at 1.5 W m-2) and predicted a photochemical quantum yield of >90%. Exogenous donors and acceptors had no effect on the signal amplitudes indicating that energy storage is the result of charge separation between endogenous components. Fe(CN)6 -3 oxidation of P700 and dithionite-induced reduction of acceptors FA-FB inhibited energy storage. These data are compatible with the hypothesis that energy storage in PS I arises from charge separation between P700 and Fe-S centers FA-FB that is stable on the time scale of the photoacoustic modulation. High intensity background light (160 W m-2) caused an irreversible loss of energy storage and correlated with a decrease in oxidizable P700; both are probably the result of high light-induced photoinhibition. By analogy to the low fluorescence yield of PS I, the low signal-to-noise ratio in these preparations is attributed to the short lifetime of Chl singlet excited states in PS I-40 and its indirect effect on the yield of thermal emission.Abbreviations FFT fast Föurier transform - HA hydroxyapatite - I50 half saturation intensity for energy storage - PA photoacoustic - PS photosystem - PS I-40 photosystem I reaction center/core antenna complex containing about 40 Chl a/P700 - 201-1 photoacoustic energy storage signal - S/N signal-to-noise  相似文献   

13.
After saturating light illumination for 3 h the potential photochemical efficiency of photosystem Ⅱ (PSII) (FJF,, the ratio of variable to maximal fluorescence) decreased markedly and recovered basically to the level before saturating light illumination after dark recovery for 3 h in both soybean and wheat leaves, indicating that the decline in FJ/Fm is a reversible down-regulation. Also, the saturating light illumination led to significant decreases in the low temperature (77 K) chlorophyll fluorescence parameters F685 (chlorophyll a fluorescence peaked at 685 nm) and F685/F735 (F735, chlorophyll a fluorescence peaked at 735 nm) in soybean leaves but not in wheat leaves. Moreover, trypsin (a protease) treatment resulted in a remarkable decrease in the amounts of PsbS protein (a nuclear gene psbS-encoded 22 kDa protein) in the thylakoids from saturating light-illuminated (SI), but not in those from darkadapted (DT) and dark-recovered (DRT) soybean leaves. However, the treatment did not cause such a decrease in amounts of the PsbS protein in the thylakoids from saturating light-illuminated wheat leaves. These results support the conclusion that saturating light illumination induces a reversible dissociation of some light-harvesting complex Ⅱ (LHClI) from PSII reaction center complex in soybean leaf but not in wheat leaf.  相似文献   

14.
Net photosynthetic rate of yellow upper leaves (UL) of Ligustrum vicaryi was slightly, but not significantly higher than that of green lower leaves (LL). Diurnally, maximum photochemical efficiency of photosystem 2, PS2 (Fv/Fm) of LL did not significantly decline but the UL showed fairly great daily variations. Yield of PS2 of UL showed an enantiomorphous variation to the photosynthetically active radiation and was significantly lower than in the LL. Unlike Fv/Fm, the efficiency of energy conversion in PS2 and both non-photosynthetic and photosynthetic quenching did not differ in UL and LL. Significant differences between UL and LL were found in contents of chlorophyll (Chl) a, b, and carotenoids (Car) and ratios of Chl a/b, Chl b/Chl (a+b), and Car/Chl (a+b). Leaf colour dichotocarpism in L. vicaryi was mainly caused by different photon utilization; sunflecks affected the LL.  相似文献   

15.
Seeni S  Gnanam A 《Plant physiology》1982,70(3):815-822
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.

  相似文献   

16.
Long-term (30 d) effects of 100, 200, 300, and 400 mM NaCl on photosystem 2 (PS 2)-mediated electron transport activity and content of D1 protein in the thylakoid membranes of chrysanthemum (Dendranthema grandiflorum) cultured in vitro at low irradiance 20 μmol(photon) m−2 s−1 were investigated. 100 mM NaCl increased contents of chlorophylls (Chl) a and b, carotenoids (Car; xanthophylls + carotenes), and the ratio of Chl a/b, and Car/Chl a+b. However, further increase in NaCl concentration led to the significant reduction in the contents of Chl a, and Chl b, and increase in the ratio of Chl a/b and Car/Chl a+b. NaCl treatment decreased the PS 2-mediated electron transport activity and contents of various thylakoid membrane polypeptides including D1 protein.  相似文献   

17.
A comparative study of reciprocal conversions of chlorophylls a and b (Chl aand Chl b) in etiolated and post-etiolated rye seedlings (Secale cereale L.) was performed. The production of these pigments was initiated by infiltration of exogenous chlorophyllides a and b (Chlide a and b). It was shown that Chlide b, when infiltrated into etiolated rye seedlings, was esterified, producing Chl b. A major portion of Chl b (more than 80%) was transformed into Chl aduring long-term seedling dark exposure. The high rate of Chl b conversion into Chl a in the pool of pigments of exogenous origin was also observed during the lag-phase when there was no chlorophyll formation from endogenous precursors. The infiltration of Chlide a resulted in Chl a formation. The efficiency of its conversion into Chl b was low (about 1%) in the etiolated seedlings but increased during their greening. In the post-etiolated seedlings infiltrated with Chlide b, which were preliminary illuminated for 6–12 h, the Chl /Chl a ratio was almost similar in the pools of pigments synthesized from both exogenous and endogenous precursors. The rates of direct and reverse reactions responsible for the interconversion of Chl aand Chl b depended on the stage of the formation of the photosynthetic apparatus during greening of etiolated seedlings, when the particular structural components are formed in a definite sequence.  相似文献   

18.
Two fractions of the light fragments enriched in the photosystem I (PSI) complexes were obtained from pea (Pisum sativum L.) thylakoids by digitonin treatment and subsequent differential centrifugation. The ratio of chlorophyll a to chlorophyll b, chlorophyll/P700 spectra of low-temperature fluorescence, and excitation spectra of long-wave fluorescence were measured. These characteristics were shown to be different due to variation in the size and composition of the light-harvesting antenna of PSI complexes present in the particles obtained. The larger antenna size of one of the fractions was related to the incorporation of the pool of light-harvesting complex II (LHCII). A comparison with the data available allowed us to identify these particles as fragments of intergranal thylakoids and end membranes of granal thylakoids. The suggestion that an increase in the PSI light-harvesting antenna in intergranal thylakoids is related to the attachment of phosphorylated LHCII is discussed.  相似文献   

19.
In grapevine (Vitis vinifera L.) leaf chlorophyll (Chl) a and Chl b and carotenoid contents were higher in plants grown at low photon flux densities (PFD) than in those grown at medium and high PFD. The highest Chl a variable to maximum fluorescence ratio Fv/Fm was observed in plants grown at medium PFD while the minimum fluorescence F0 was highest in those at high PFD. In isolated thylakoids, both high and low PFD caused marked inhibition of whole chain and photosystem 2 (PS2) activities. The artificial exogenous electron donor diphenyl carbazide significantly restored the loss of PS2 activity in low PFD leaves.  相似文献   

20.
Białek-Bylka  G.E.  Sofrová  D.  Szurkowski  J.  Skwarek  R.  Sopko  B.  Manikowski  H. 《Photosynthetica》2000,38(1):143-148
Pigment-protein complexes enriched in photosystem 1 (PS1) and, for comparison, enriched in photosystem 2 (PS2) were isolated from the cyanobacterium Synechococcus elongatus Nag. f. thermalis Geitl. They were immobilized and oriented in the polyvinyl alcohol (PVA) films, and studied by linear dichroism (LD), fluorescence polarization (FP), photoacoustic spectroscopy (PAS), and polarized photoacoustic spectroscopy (PAS and PAS). The LD signal of -carotene in the region with maximum at 500 nm was positive in the PS1 complex. The maximum value of fluorescence polarization (FP) in the measured photosynthetic pigment region was 1.25 and was similar to higher plant values. Carotenoids exhibited different efficiencies of thermal deactivation (max. at 500 nm) in PS1 and PS2. The thermal deactivation efficiency of carotenoids in comparison with that of chlorophyll (Chl) a at its red absorbance maximum was much higher in PS1 than in PS2 complexes. Cyanobacterial complexes did not contain Chl b, interpretation of the LD, PAS, and FP results is thus easier and can be compared with PS1 and PS2 values of higher plants, especially with Chl b-less mutant values.  相似文献   

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