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1.
Photosynthetically competent chloroplasts were isolated fromcells of Euglena gracilis Z grown photoautotrophically in 1.5%CO2. The isolated chloroplasts were intact and substantiallyfree from cytosolic, mitochondrial and microbody materials.The effects of some compounds on the activity of photosynthetic14CO2 fixation were examined. The optimal pH and sorbitol concentrationwere 8.0 and 0.33 M, respectively. The chloroplasts requireda high level of P, (5 to 20 mM) for the maximal rate of photosynthesis.They were insusceptible to 10 mM of free Mg2+. ATP, ADP andAMP at 1 to 5 mM notably stimulated photosynthesis, althoughhigh concentrations of AMP were unfavorable. In the assay mediumdeveloped for this study, the chloroplasts exhibited photosyntheticactivity of 120µmoles-mg1 Chl-h1 at 30?C. Chloroplasts could also be isolated from cells grown under ordinaryair. The rate of photosynthetic 14CO2 fixation at 1 mM NaHl4CO3was higher in these chloroplasts than in those isolated fromcells grown in 1.5% CO2, whereas at 10 mM NaHl4CO3, the ratesof the two types of chloroplasts were nearly the same. Theseresults suggest that the CO2 concentration given during growthof the algal cells affects the affinity for dissolved inorganiccarbon at the chloroplast level. (Received March 30, 1987; Accepted August 17, 1987) 相似文献
2.
In cells of cyanobacterium Anabaena variabilis grown under ordinaryair (low-CO2 cells), the transport of both CO2 and HCO3was significantly enhanced by Na+. This effect was pronouncedas the external pH increased. When low-CO2 cells were treatedwith an inhibitor of carbonic anhydrase (CA), only CO2 transportbut not HCO3 transport, was inhibited. The initial rateof photosynthetic carbon fixation as a function of the concentrationof internal inorganic carbon (IC) was practically the same irrespectiveof whether CO2 or HCO3 was externally supplied. Theseresults suggest that IC is actively transported through theplasma membrane in a form of HCO3 probably by some transporterand that the transmembrane Na+ gradient is involved in thisIC transport system. Free CO2 may be hydrated by CA to HCO3and then transported to the cells by this transporter. On the other hand, CO2 is actively taken up by cells grown withair containing 5% CO2 (high-CO2 cells) though the enhancingeffect of Na+ was much smaller in high- CO2 cells than in low-CO2cells. The initial rate of fixation as a function of internal IC concentrationindicated that the rate of the carboxylation reaction of accumulatedIC is higher in I0W-CO2 cells than in high-CO2 cells. The studieswith ethoxyzolamide indicated that even in low-CO2 cells, CAdoes not function inside Anabaena cells. These results suggestthat inside the low-CO2 cells of Anabaena, some mediator(s)facilitates the transport of IC to RuBPCase. (Received January 23, 1987; Accepted April 24, 1987) 相似文献
3.
Acidic inorganic phosphate (Pi) pool (pH around 6) was detected besides the cytoplasmic pool in intact cells of Chlorella vulgaris 11h by 31P-in vivo nuclear magnetic resonance (NMR) spectroscopy. It was characterized as acidic compartments (vacuoles) in combination with the cytochemical technique; staining the cells with neutral red and chloroquine which are known as basic reagents specifically accumulated in acidic compartments. Under various conditions, the results obtained with the cytochemical methods were well correlated with those obtained from in vivo NMR spectra; the vacuoles were well developed in the cells at the stationary growth phase where the acidic Pi signal was detected. In contrast, cells at the logarithmic phase in which no acidic Pi signal was detected contained only smaller vesicles that accumulated these basic reagents. No acidic compartment was detected by both cytochemical technique and 31P-NMR spectroscopy when the cells were treated with NH4OH. The vacuolar pH was lowered by the anaerobic treatment of the cells in the presence of glucose, while it was not affected by the external pH during the preincubation ranging from 3 to 10. Possible vacuolar functions in unicellular algae especially with respect to intracellular pH regulation are discussed.Non-standard abbreviations EDTA
ethylenediaminetetraacetic acid
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- MDP
methylene diphosphonic acid
- NMR
nuelear magnetic resonance
- PCA
perchloric acid
- PCV
packed cell volume
- Pi
inorganic phosphate
- Pic
sytoplasmic inorganic phosphate
- Piv
vacuolar inorganic phosphate
- ppm
parts per million
- SP
sugar phosphates
- TCA
trichloroacetic acid 相似文献
4.
Summary The pyrenoid is a protein complex in the chloroplast stroma of eukaryotic algae. After the treatment with mercury chloride, pyrenoids were isolated by sucrose density gradient centrifugation from cell-wall less mutant cells, CW-15, as well as wild type cells, C-9, of unicellular green algaChlamydomonas reinhardtii. Pyrenoids were characterized as a fraction whose protein/chlorophyll ratio was very high, and also examined by Nomarski differential interference microscopy. Most of the components consisted of 55 kDa and 16 kDa polypeptides (11) which were immunologically identified as the large and small subunit of RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase) protein, respectively. Some minor polypeptides were also detected. Substantial amount of RuBisCO protein is present as a particulate form in the pyrenoid in addition to the soluble form in algal chloroplast stroma.Abbreviations BPB
bromophenol blue
- DAB
3,3-diaminobenzidine
- DTT
dithiothreitol
- ELISA
enzyme-linked immunosorbent assay
- High-CO2 cells
cells grown under air enriched with 4% CO2
- Low-CO2 cells
cells grown under ordinary air (containing 0.04% CO2)
- NP-40
nonionic detergent (Nonidet) P-40
- PAGE
polyacrylamide gel electrophoresis
- PAP
peroxidase-antiperoxidase conjugate
- RuBisCO
ribulose-1,5-bisphosphate carboxylase/oxygenase
- RuBP
ribulose-1,5-bisphosphate
- SDS
sodium dodecylsulfate 相似文献
5.
Tominaga Yoshito; Kuchitsu Kazuyuki; Katsuhara Maki; Tazawa Masashi; Miyachi Shigetoh 《Plant & cell physiology》1991,32(2):261-268
Rotational streaming of the cytoplasm including chloroplastswas induced by L-histidine, as well as by light, on the anticlinalface of leaf cells of Egeria densa. In the case of treatmentwith L-histidine some of the chloroplasts remained stationaryon the periclinal face of cells after rotational cytoplasmicstreaming was initiated. However, these chloroplasts were easilydislodged and translocated to the centrifugal end of the histidine-treatedcells by application of a centrifugal force that barely affectedthe location of chloroplasts in cells incubated in the darkwithout L-histidine. This result indicates that the anchoringof chloroplasts was weakened by L-histidine. Thus only the releaseof chloroplasts from anchoring was not enough for initiationof their streaming. The cytoplasmic pH (pHc) and vacuolar pH(pHv) were noninvasively monitored by in vivo 31P-nuclear magneticresonance (NMR) spectroscopy. Compared with the dark controlvalue, both illumination and treatment with L-histidine increasedthe pHc by 0.3 units. In contrast, pHv changed only a littlewith both illumination and treatment with L-histidine. Releaseof chloroplasts from anchoring and initiation of cytoplasmicstreaming are discussed in relation to the increase in pHc inducedby both light and L-histidine.
4 Present address: Department of Cell Biology, National Instituteof Agrobiological Resources, Kannondai, Tsukuba, Ibaraki, 305Japan
5 Present address: Marine Biotechnology Institute Co., Ltd.,Head Office, 2-35-10 Hongo, Bunkyo-ku, Tokyo, 113 Japan (Received July 16, 1990; Accepted December 20, 1990) 相似文献
6.
7.
Carbonic anhydrase (CA) activity was detected in homogenatesfrom Anabaena variabilis ATCC 29413, M-2 and M-3, but not inthe suspension of the intact cells. Activity was higher in cellsgrown in ordinary air (low-CO2 cells) than in those grown inair enriched with 24% CO2 (high-CO2 cells). Fractionationby centrifugation indicated that the CA from A. variabilis ATCC29413 is soluble, whereas both soluble and insoluble forms existin A. variabilis M-2 and M-3. The addition of dithiothreitoland Mg2 $ greatly decreased the CA activity of A. variabilisATCC 29413. The specific activity of the CA from A. variabilis ATCC 29413was increased ca. 200 times by purification with ammonium sulfate,DEAE-Sephadex A-50 and Sephadex G-100. Major and minor CA peaksin Sephadex G-100 chromatography showed respective molecularweights of 48,000 and 25,000. The molecular weight of the CAdetermined by polyacrylamide disc gel electrophoresis was 42,000?5,000.The activity of CA was inhibited by ethoxyzolamide (I50=2.8?10-9M), acetazolamide (I50=2.5?10-7 M) and sulfanilamide (I50=2.9?10-6M). (Received January 5, 1984; Accepted April 26, 1984) 相似文献
8.
Analysis of products formed in Chlorella vulgaris 11 h cellsduring photosynthesis in air containing 3,000 ppm 14CO2 at varioustemperatures revealed that the level of 14C-starch was maximumaround 2024?C and decreased with further rise in temperatureuntil 40?C, while 14C-sucrose greatly increased at temperaturesabove about 28?C. Elevating the temperature from 20 to 38?Cduring photosynthetic 14CO2 fixation resulted in a remarkabledecrease in 14C in starch and a concomitant increase in 14Cin sucrose. This conversion of starch to sucrose when shiftingthe temperature from 20 to 38?C proceeded even in the dark.Hydrolysis of sucrose by rß-fructosidase showed that,irrespective of the experimental conditions, the radioactivitiesin sucrose were equally distributed between glucose and fructose.The enhancement of starch degradation with temperature risewas more remarkable than that of the activity of ribulose bisphosphatecarboxylase from the same cells. When Chlorella cells whichhad been preloaded with 14C-starch after photosynthesis for30 min at 20?C were incubated in the dark for an additional30 min at 20?C, 14C-starch was degraded by only about 4%. However,the values after 30-min dark incubation at 28, 32, 36 and 40?Cwere increased by about 10, 19, 36 and 50%, respectively. Duringthe temperature-dependent conversion of starch to sucrose, nosignificant amount of radioactivity accumulated in free glucoseand maltose. (Received October 27, 1981; Accepted January 9, 1982) 相似文献
9.
Chlorella vulgaris 11h cells grown in air enriched with 4% CO2(high-CO2 cells) had carbonic anhydrase (CA) activity whichwas 20 to 90 times lower than that of algal cells grown in ordinaryair (containing 0.04% CO2, low-CO2 cells). The CO2 concentrationduring growth did not affect either ribulose 1,5-bisphosphate(RuBP) carboxylase activity or its Km for CO2. When high-CO2 cells were transferred to low CO2 conditions,CA activity increased without a lag period, and this increasewas accompanied by an increase in the rate of photosynthetic14CO2 fixation under 14CO2-limiting conditions. On the otherhand, CA activity as well as the rate of photosynthetic 14CO2fixation at low 14CO2 concentrations decreased when low-CO2cells were transferred to high CO2 conditions. Diamox, an inhibitor of CA, at 0.1 mM did not affect photosynthesisof low-CO2 cells at high CO2 concentration (0.5%). Diamox inhibitedphotosynthesis only under low CO2 concentrations, and the lowerthe CO2 concentration, the greater was the inhibition. Consequently,the CO2 concentration at which the rate of photosynthesis attainedone-half its maximum rate (Km) greatly increased in the presenceof this inhibitor. When CO2 concentration was higher than 1%, the photosyntheticrate in low-CO2 cells decreased, while that in high-CO2 cellsincreased. Fractionation of the low-CO2 cells in non-aqueous medium bydensity showed that CA was fractionated in a manner similarto the distribution of chlorophyll and RuBP carboxylase. These observations indicate that CA enhances photosynthesisunder CO2-limiting conditions, but inhibits it at CO2 concentrationshigher than a certain level. The mechanism underlying the aboveregulatory functions of CA is discussed.
1This work was reported at the International Symposium on PhotosyntheticCO2-Assimilation and Photorespiration, Sofia, August, 1977 (18).Requests for reprints should be addressed to S. Miyachi, RadioisotopeCentre, University of Tokyo, Bunkyo-ku, Tokyo 113, Japan. (Received December 11, 1978; ) 相似文献
10.