Interaction of Benzoquinones with QA- and QB- in Oxygen-Evolving Photosystem II Particles from the Thermophilic Cyanobacterium Synechococcus elongatus

The interactions of benzoquinones with the reduced forms ofthe bound plastoquinone acceptors, Q_A and Q_B, were studied withoxygen-evolving photosystem II (PS II) particles from the thermophiliccyanobacterium Synechococcus elongatus, which largely lack poolplastoquinone molecules [Takahashi and Katoh (1986) Biochim.Biophys. Acta 845: 183]. Oxygen evolution in the presence ofvarious electron acceptors was determined and flash-inducedchanges in absorbance in the blue region were analyzed in termsof difference spectra, dependence on the concentration of benzoquinoneand on temperature, and sensitivity to 3-(3,4-dichlorophenyl)-1,1-dimethylurea(DCMU). The more hydrophobic the quinone molecule, the higherwas the rate of oxygen evolution, and the maximum rate of 3,000µmoles O₂.(mg chlorophyll)^–1.h^–1 was recordedin the presence of phenyl- and dichloro-p-benzoquinones. DCMUinhibited oxygen evolution by more than 95%. However, spectrophotometricstudies revealed that, even though electrons were transferredto benzoquinones predominantly via the direct oxidation of by added benzoquinones occurred in such a way as to indicate thatabout 40% of PS II reaction centers were not associated withfunctional Q_B sites. was very stable in the presence of ferricyanide. However, benzoquinonesinduced the slow oxidation of . The characteristics of the benzoquinone reductioin in thePS II preparation is discussed. ¹Present address: Department of Life Sciences, Faculty of Science,Himeji Institute of Technology, Shosha 2167, Himejishi, Hyogo-ken,671-22 Japan (Received May 8, 1990; Accepted August 14, 1990)     

The influence of temperature and light on the upper limit of Microcystis aeruginosa production in a hypertrophic reservoir

Primary production was measured for 7 years, using the in situ¹⁴C-method in hypertrophic Hartbeespoort Dam, South Africa,to examine the influence of light and water temperature on theupper limit of Microcystis aeruginosa production. Water temperaturesvaried from 11 to >25°C and chlorophyll concentrationsreached 6500 mg m^–3. The maximum volumetric rate of production(A_max) was 12->8800 mg C m^–3 h^–1 with areal productions(A) of 69->3300 mg C m^–2 h^–1 for euphotic zonedepths of <0.5–8.4 m. The intrinsic parameters of phytoplanktonproduction (, A_max/B, I_k) indicated that the phytoplankton populationwas adapted to high light levels. Both A_max/B and I_k were correlatedwith temperature. Under optimal conditions, , the theoreticalupper limit of A, was calculated to be 2.8 g Cm^–2 h^–1,while the measured rate was 2.5 g Cm^–2 h^–1. Measuredareal rates exceeding were overestimated due to methodologicalproblems when working with Microcystis scums. Light and watertemperature interacted to yield high production rates: watertemperature through its direct effect on photosynthetic ratesand indirectly in the formation of diurnal mixed layers; lightindirectly through water temperature and directly through itsattenuation and induction of light-adapted physiology in Microcystis.     

Simultaneous changes in cell quotas of microcystin, chlorophyll a, protein and carbohydrate during different growth phases of a batch culture experiment with Microcystis aeruginosa

The cell quotas of microcystin (Q_mcyst), protein (Q_prot), chlorophylla (Q_chloro) and carbohydrate (Q_carbo), as well as the net productionrates of these parameters, were determined during the exponentialand stationary phases in nine batch cultures of Microcystisaeruginosa (CYA 228) at light regimes from 33 to 53 µmolphotons m^–2 s^–1. The following results were obtained.(i) A parallel pattern was found in the changes of Q_mcyst, Q_prot,Q_chloro and Q_carbo during the entire growth cycle and significantcorrelations were recorded between Q_mcyst and Q_prot, Q_chloroand Q_carbo. (ii) The net microcystin production rate (µ_mcyst)was positively correlated with the specific cell division rate(µ_c), the chlorophyll production rate (µ_chloro)and the protein production rate. (iii) A significant inverselinear relationship was found between µ_c and Q_mcyst, i.e.cultures with a positive µ_c had a Q_mcyst between 110 and400 fg microcystin cell^–1, while declining cultures hadQ_mcyst values >400 fg microcystin cell^–1. Maximum variationin Q_mcyst within cultures was 3.5-fold. Collectively, the resultsshow that cells produced microcystin at rates approximatingthose needed to replace losses to daughter cells during divisionand that microcystin was produced in a similar way to proteinand chlorophyll, indicating a constitutive microcystin production.     

Reaction of Reconstituted Acceptor Quinone and Dynamic Equilibration of Electron Transfer in the Photosystem I Reaction Center

Electron transfer mechanism in the spinach photosystem I reactioncenter that contains artificial quinones in place of phylloquinone(2-methyl-3-phytyl-1,4-naphthoquinone, vitamin K₁) as the secondaryelectron acceptor, Q_ø (or A₁) was discussed. (1) Mostof the reconstituted quinones oxidized the primary acceptorchlorophyll a, A⁰, at a rate rapid enough to compete againstthe charge recombination between A₀^– and the oxidizeddonor chlorophyll P700⁺. (2) The pathway of electron transferfrom the semiquinone^– varied depending on the redox potentialvalue of each semiquinone^– /quinone couple. Low potentialquinones reduced the tertiary acceptor iron-sulfur center, F_x,while the high potential ones reduced P700⁺ directly with a200-µs halftime. (3) The E_m value of each semiquinone^–/quinone couple in situ in the reaction center was estimatedto be shifted by about 0.3 volt to the negative side from theirhalf wave redox potential values that were measured polarographicallyin dimethylformamide. The shift seems to represent the acceptorproperty of the protein environment at the Q_ø site. (4)The E_m of reconstituted phylloquinone was estimated to be 50–80mV more negative than that of F_x. (5) The mechanism of efficientelectron transfer in the reaction center was discussed basedon the dynamic equilibria between the electron transfer componentsand on the estimated E_m values. (Received April 9, 1994; Accepted July 7, 1994)     

Properties of Chlamydomonas Photosystem II Core Complex with a His-Tag at the C-Terminus of the D2 Protein

A His-tagged PSII core complex was purified from recombinantChlamydomonas reinhardtii D2-H thylakoids by single-step Ni²⁺-affinitycolumn chromatography and its properties were partially characterizedin terms of their PSII functions and chemical compositions.The PSII core complex that has a His-tag extension at the C-terminusof the D2 protein evolved oxygen at a high rate of 2,400 µmol(mg Chl)^–1h^–1 at the optimum pH of 6.5 with ferricyanideand 2,6-dichlorobenzoquinone as electron acceptors in the presenceof Ca²⁺ as an essential cofactor, and approximately 90% of theactivity was blocked by 10 µM DCMU. The core complex exhibitedthe thermoluminescence Q-band but not the B-band regardlessof the presence or absence of DCMU, although both bands wereobserved in the His-tagged thylakoids. The core complex wasfree from PSI and contained one Y_D, Tyr 160 of the D2 protein,four Mn atoms, two cytochrome b-559, about 46 Chl a molecules,and probably one Q_A, the primary acceptor quinone of PSII. Itwas inferred from these results that His-tagging at the C-terminusof the D2 protein does not affect the functional and structuralintegrity of the PSII core complex, and that the ‘His-tagstrategy’ is highly useful for biochemical, physicochemical,and structural studies of Chlamydomonas PSII. (Received October 22, 1998; Accepted December 25, 1998)     

Limitations to net photosynthesis as affected by nitrogen status in jack pine (Pinus banksiana Lamb.) seedlings

Relative limitations of nitrogen (N) status on the processescontributing to photosynthetic rate (A) were investigated. Jackpine {Pinus banksiana Lamb.) seedlings from seeds grown in sandculture were supplied with four different N treatments for 6weeks, which resulted in a needle N content ranging from 50–85mmol m^–2 (14–32 mg g^–1 dry weight). Leaf gasexchange at varying CO₂ levels was measured and limitationson A₃₅₀ (A at ambient CO₂ level) caused by finite, limitingcarboxylation efficiency (c.e.), maximum A (A_max)and stomatalconductance were estimated from an analysis of the responseof A to internal CO₂ concentration. Although c.e. and A_max decreasedlinearly with the decline in needle N, the magnitudes of theirchanges relative to A₃₅₀ differed. A_max varied with A₃₅₀ andalways exceeded A₃₅₀ by 37–38% c.e., however, declinedfaster than A₃₅₀, as needle N level decreased. Consequently,relative limitation on A₃₅₀ caused by inefficient A_max remainedconstant, but limitations caused by c.e. increased by 10–15%at low N levels. In contrast, the limitation by stomatal conductancedeclined initially, but remained stable when N content droppedbelow 75 mmol m^–2. The results suggest: (1) a decreasein biochemical capacity, but not stomatal conductance, contributedto the reduction of A₃₅₀ induced by N-deficiency in jack pineseedlings; and (2) the capacity of carboxylation appeared tobe impaired more than that of electron transport and/or photophosphorylationand its reduction may be the major reason for the reductionin A₃₅₀. Key words: A–C_i analysis, carboxylation efficiency, electron transport, nitrogen deficiency, stomatal conductance     

pH and Temperature Dependence of Tyrosine Z Decay Kinetics in Tris-Treated PSII Particles Studied by Time-Resolved EPR

The decay kinetics of tyrosine Z (Y_z) in Tris-treated PhotosystemII particles were measured by time-resolved EPR at differentpH values (pH 5.5 to 7.5) between 230 and 297 K. Y_z inducedby laser flashes decayed in a biphasic wav; t values were about100 ms in the fast phase and about 1 s in the slow one, respectively,at room temperature. The fast phase was attributed to a recombinationof charges on Y_z and Q^–_A. The activation energies forthe reaction of Y_z with Q^–_A between 245 and 297 K havebeen estimated to be more than 38 kJ mol^–1 at pH (>6.5)and less than 32 kJ mol^–1 at pH (<6.0), respectively.The activation energy gap between high pH (>6.5) and lowpH (<6.0) ranges was found to be E=4.4 kJ mol^–1. Judgingfrom the fact that pH 6.2 appears to be the border between thehigh and low pH regions, we suggest that the kinetics of Y_zis strongly influenced by the dissociation of the nearby histidineresidue. (Received February 21, 1997; Accepted June 24, 1997)     

Physiological evaluation of temperature effect on the growth processes of the mysid, Neomysis intermedia Czerniawsky

Ingestion, respiration, and molting loss rates were measuredover the 3 – 29°C range in Neomysis intermedia. Weightspecific rates of these physiological processes ranged from2 to 140% body C day^–1 for ingestion, from 2 to 15% bodyC day^–1 for respiration, and from 0.1 to 5% body C day^–1for molting loss. All weight-specific rates showed a logarithmicdecrease with a logarithmic increase in body weight, and a logarithmicincrease with a linear increase in temperature below 20 or 25°C.The effect of temperature, however, was different between thephysiological rates, with a large temperature dependency foringestion (Q₁₀ = 2.6 –3.9) and molting loss (Q¹⁰ = 2.9– 3.6) and a moderate temperature dependency for respiration(Q₁₀ = 1.9 – 2.1). Calculated assimilation efficiencychanged with body size, but was constant over the temperaturerange examined. Allocation of assimilated materials varied witha change in temperature, reflecting the different temperaturedependence between physiological processes. It was deduced thatthe strong temperature dependency of the growth rate in N. intermediaobserved in the previous studies resulted from the large temperatureeffect on ingestion and assimilation rates, superimposed bythe different allocation of assimilated materials. ¹Present address: Department of Botany, University of Tokyo,Hongo, Tokyo 113, Japan     

Planktonic primary production in the German Wadden Sea

By combining weekly data of irradiance, attenuation and chlorophylla concentrations with photosynthesis (P) versus light intensity(E) curve characteristics, the annual cycle of planktonic primaryproduction in the estuarine part of the Northfrisian WaddenSea was computed for a 2 year period. Daily water column particulategross production ranged from 5 to 2200 mg C m^–2 day^–1and showed a seasonal pattern similar to chlorophyll a. Budgetcalculation yielded annual gross particulate primary productionsof 124 and 176 g C m^–2 year^–1 in 1995 and 1996,respectively. Annual amounts of phytoplankton respiration, calculatedaccording to a two-compartment model of Langdon [in Li,W.K.W.and Maestrini,S.Y. (eds), Measurement of Primary Productionfrom the Molecular to the Global Scale. International Councilfor the Exploration of the Sea, Copenhagen, 1993, pp. 20–36],and dissolved production in 1996, were both in the range of24–39 g C m^–2 year^–1. Annual total net productionwas thus very similar to particulate gross production (127 and177 g C m^–2 year^–1 in 1995 and 1996, respectively).Phytoplankton growth was low or even negative in winter. Inspring and summer, production/biomass (Pr/B) ratios varied from0.2 up to 1.7. Phytoplankton growth during the growth seasonalways surpassed average flushing time in the area, thus underliningthe potential of local phytoplankton bloom development in thispart of the Wadden Sea. The chlorophyll-specific maximum photosyntheticrate (P^B_max) ranged from 0.8 to 9.9 mg C mg^–1 Chl h^–1and was strongly correlated with water temperature (r² = 0.67).By contrast, there was no clear seasonal cycle in ^B, which rangedfrom 0.007 to 0.039 mg C mg^–1 Chl h^–1 (µmolphotons m^–2 s^–1)^–1. Its variability was muchless than P^B_max and independent of temperature. The magnitudeand part of the variability of P^B_max and ^B are presumably causedby changes in species composition, as evidenced from the rangeof these parameters found among 10 predominant diatom speciesisolated from the Wadden Sea. The ratio of average light conditionsin the water column (E_av) to the light saturation parameterE_k indicates that primary production in the Wadden Sea regionunder study is predominantly controlled by light limitationand that nutrient limitation was likely to occur for a few hoursper day only during 5 (dissolved inorganic nitrogen) to 10 (PO₄,Si) weeks in the 2 year period investigated.     

Factors controlling primary production in a hypertrophic lake (Hartbeespoort Dam, South Africa)

The physical factors controlling algal primary production weredemonstrated from data collected for a hypertrophic lake. A_maxranged between 12.4 and 5916 mg C m^–3 h^–1. Arealrates (A) varied between 46.9 and 3381 mg C m^–2 h^–1.The factors permitting and controlling production were subjectivelyseparated into two categories. In category 1, nutrients (N +P), which were in overabundance, permitted large standing cropsof Microcystis aeruginosa to develop (>1000 µg chla 1^–1). Wind patterns determined the dramatic spatialand temporal changes in algal standing crop which could dropto 2.7 µg chl a 1^–1. In category 2 were the factorswhich affected the rate processes. The buoyancy mechanism ofMicrocystis usually kept the alga in the euphotic zone. A powerrelationship (r = 0.92, n = 54) between A and A_max/_min showedthat with increasing phytoplankton vertical stratification,A_max was increasingly important in the integral. The saturationparameter I_K and photosynthetic capacity were temperature dependent.Variations of A were significantly related to changes in watercolumn stability (g cm cm^–2) because both axes of thephotosynthesis depth-profile were affected by stability changes.     

Phytotoxicity of Phthalate Plasticisers: 2. SITE AND MODE OF ACTION

The effects of phthalate esters on chlorophyll a₂ fluorescencein radish plants (Raphanus sativus L. cv. Cherry Belle) wereexamined Fluorescence yield was increased in those plants exposedto an aerial concentration of 120 ng dm^–3 dibutyl phthaiatc(DBP) at a rate of 3.0 dm³ min^–1 for 13 d. Comparisonof fluorescence enhancement ratios and F_red/F_ox, suggests thatDBP inhibits photosynthesis in radish plants at a site afterQ_A. Both DBP and diisobutyl phthalate (DIBP) strongly inhibiteduncoupled (PS2+PS1) electron transport rates in thylakoids isolatedfrom spinach. At a chlorophyll concentration of 10 µgcm^–3 the concentrations of DBP and DIBP exhibiting 50%inhibition were 44 mmol m^–3 and 42 mmol m^–3 respectively.Basal electron transport rates were also inhibited, with 87mmol m^–3 of DBP or DIBP producing 50% inhibition. Measurementof photosystcm 1 activity suggested that the main site of actionof these phthalates was localized at a site near the reducingside of photosystem 2. Key words: Phthalate, plasticiser, chlorophyll, fluorescence, photosynthesis, inhibition     

Production of Oikopleura longicauda (Tunicata: Appendicularia) in Toyama Bay, southern Japan Sea

Oikopleura longicauda occurred throughout the year in ToyamaBay, southern Japan Sea, and analysis of its size compositionand maturity revealed that reproduction was continuous overtheyear. Somatic growth production (P_g) varied with season from0.03 to 103 mg carbon (C) m^–2day^–1 (annual P_g 4.5g C m^–2), and house production (P_e) from 0.11 to 266 mgC m^–2 day^–1 (annualP_e 11.3 g C m^–2). The annualP_g/B ratio was 176. Compared with production data of some predominantzooplankton species in Toyama Bay, it is suggested that despitetheir smaller biomass, appendicularians are an important secondaryproducer.     

Mechanisms of Electron Flow through the QB Site in Photosystem II. 3. Effects of the Presence of Membrane Structure on the Redox Reactions at the QB Site

Reduction kinetics of chloro- or methyl-substituted benzoquinones(BQs) in spinach PSII membrane fragments or n-heptyl-rß-D-thioglucoside-extractedPSII core complexes (HTG-PSII) was studied and compared to thatin cyanobacterial PSII core complexes [Satoh et al. (1995) PlantCell Physiol. 36: 597]. It was found that the BQs accept electronsat two sites (the Q_B and PQ sites) in the both spinach preparationsas in the cyanobacterial preparation. Maximum turnover rates(V_max) and binding affinities (K_m) of the two sites were estimated.Comparison of the values in PSII membrane fragments with thosein HTG-PSII showed that removal of the membrane structure orlight-harvesting chlorophyll a/b protein complexes from thePSII core complexes had little effect on the characteristicsof the Q_B site, indicating that the HTG-PSII have the intactQ_B site and are good materials to study the site. The K_m andV_max values were comparable to those in the cyanobacterial preparation. Low affinity (high K_m values) and high V_max values of methyl-substitutedBQs to the Q_B site and almost the same rate of intrinsic electronflow through the Q_B site in the both spinach preparations furthersupport the hypothesis that the plastoquinone (PQ) moleculeat the Q_B site is not replaced by another PQ molecule but, afterreduction, only its head group goes out of the Q_B pocket, andcomes back after transferring the electrons and protons to afree PQ molecule [Satoh et al. (1993) Z. Naturforsch. 48c: 174]. (Received May 2, 1996; Accepted July 26, 1996)     

The NADP$-specific isocitrate dehydrogenase of Rhodospirillum rubrum

The NADP^$-specific isocitrate dehydrogenase was partially purifiedfrom photosynthetically-grown Rhodospirillum rubrum. The pHoptimum is between 7.5 and 9.0 in phosphate buffer. The apparentKm is 3.1x10^–5 M for isocitrate, 5.1x10^–5 M forNADP^$, 1.7x10^–5 M for manganese, 1.5x10^–4 M formagnesium, and 3.5x10^–3 M for inorganic orthophosphate.Arsenate exerts a slight inhibition. The Q₁₀ between 17.5°Cand 40°C is 1.62, and the energy of activation at 25°Cis 9.74 Kcal/mole. Glyoxylate and oxalacetate cause concertedinhibition of the enzyme activity. Various nucleotides inhibitthe activity. The kinetics of inhibition by ATP was found tobe mixed type with respect to NADP^$ and isocitrate, the K_i valuesbeing 1.17x10^–3 M and 1.10x10^–3 M respectively.The inhibition between ATP and orthophosphate is competitivewith a K_i of 10^–4M. Thiol binding reagents are inhibitory;this inhibition is reversed by cysteine or reduced glutathione. (Received October 1, 1971; )     

Photosynthetic characteristics of Dinophysis norvegica Claparede & Lachmann, a red-tide dinoflagellate

The relationships between photosynthesis and photosyntheticphoton flux densities (PPFD, P-l) were studied during a red-tideof Dinophysis norvegica (July-August 1990) in Bedford Basin.Dinophysis norvegica, together with other dinoflagellates suchas Gonyaulax digitate, Ceratium tripos, contributed {small tilde}50%of the phytoplankton biomass that attained a maximum of 16.7µg Chla 1^– and 11.93 10⁶ total cells I^–1.The atomic ratios of carbon to nitrogen for D.norvegica rangedfrom 8.7 to 10.0. The photosynthetic characteristics of fractionatedphytoplankton (>30 µm) dominated by D.norvegica weresimilar to natural bloom assemblages: o (the initial slope ofthe P-l curves) ranged between 0.013 and 0.047 µg C [µgChla]^–1 h–1 [µmol m^– s^–1]^–1the maximum photosynthetic rate, p^B_m, between 0.66 and 1.85µg C [µghla]^–1 h^–1; l_k (the photoadaptationindex) from 14 to 69 µ,mol m^–2 s^–1. Carbonuptake rates of the isolated cells of D.norvegica (at 780 µmolm^–2 s^–1) ranged from 16 to 25 pg C cell^–1h^– and were lower than those for C.tripos, G.digitaleand some other dinoflagellates. The variation in carbon uptakerates of isolated cells of D.norvegica corresponded with P^B_mof the red-tide phytoplankton assemblages in the P-l experiments.Our study showed that D.norvegica, a toxigenic dinoflagellate,was the main contributor to the primary production in the bloom.     

Ecological studies on the phytoplankton of Boknafjorden, western Norway. II. Environmental control of photosynthesis

Both predicted (incubator) and measured (in situ) ¹⁴C-assimilationrates were studied from February to November 1981 at three stationsin Boknafjorden, a deep silled fjord of western Norway. Sampleswere taken from different light depths within the euphotic zone.A high degree of conformity was found between the two approaches.Daily values of carbon assimilation integrated over the euphoticzone varied between 0.05 and 1.4 g C m^–2. Yearly primaryproduction varied between stations from 82 to 112 g C m^–2(120–148 g C m^–2 when based on average light conditions).The light-saturation curve parameters ^B and P^B_max ranged from0.0056 to 0.0537 mg C mg Chla^–1 h^–1 µE^–1m² and from 0.7 to 8.5 mg C mg Chla^–1 h^–2 (in situassimilation numbers ranged from 0.9 to 9.3 mg C mg Chla^–1h^–1) respectively, which compare well with those publishedfrom the northwestern side of the Atlantic. The overall importanceof light in controlling photosynthesis throughout the year wasrevealed by the light utilization index , estimated to be 0.43mg C mg Chla^–1 E^–1 m². The maximum quantum yieldwas encountered on August 17, with 0.089 mol CE^–1. Chla/Cratios above and below 0.010 were found to be typical for shade-and light-adapted cells respectively. Assimilation numbers andgrowth rates were linearly related only when considering light-adaptedcells. Consistent with the findings of this study, the applicabilityof I_K, ^B and P^B_max as indicators of light-shade adaptation propertiesshould be questioned. Maximum growth rates were encounteredduring an autumn bloom of the dinoflagellate Gyrodinium aureolum(1.0 doublings day^–1), while 0.7–0.8 doublings day^–1were found for a winter bloom (water temperature of 2°C)of the diatom Skeletonema costatum. No unambiguous temperatureeffect on assimilation number and growth of phytoplankton couldbe recognized in Boknafjorden. A tendency towards increasedassimilation numbers coinciding with increased water columnstability was revealed. The highest P^B_max values were oftenencountered at almost undetectable nutrient concentrations.At least during summer this could be attributed to recyclingof nutrients by macro- and/or microzooplankton, responsiblefor a greater part of the primary production now being grazeddown. This study supports the convention that the depth of theeuphotic zone may extend considerably below the 1% light depth.     

Ion Fluxes in 'Isolated' Guard Cells of Commelina communis L.

Ion fluxes have been measured in ‘isolated’ guardcells of Commelina communis L. using ⁸⁶RbCl and K⁸²Br, in epidermalstrips in which all cells other than guard cells have been killedby treatment at low pH. To avoid problems of slow free spaceexchange most fluxes have been measured at pH 3.9, at whichstomata open well in K(Rb) Cl(Br) and are stable for many hours.At pH 3.9 the intracellular ⁸⁶Rb exchanged as a single compartmentwith a half-time of 2–3 h, independent of external concentration(C_o). The influx of ⁸⁶Rb rose with concentration, to a V_maxof about 23 pmol mm^–2 h^–1. The efflux curve of ⁸²Brcould be well fitted by two exponential terms, with half-timesof 38 min (independent of C_o), and 5–35 h (falling withincreasing C_o). Bromide contents of cytoplasm and vacuole (Q_cand Q_v), and fluxes at plasmalemma and tonoplast, were calculatedfrom the efflux kinetics. Over C_o 20–60 mM, as the apertureincreased from 7 µm to 17 µm, the tonoplast flux(0.5–11.5 pmol mm^–2h^–1) was always much lessthan the plasmalemma flux (7–77 pmol mm^–2 h^–1).Q_c and Q_v both increased with aperture. The increase in Q_c of10.3 pmol mm^–2 µm^–1 is adequate to accountfor the osmotic changes required to change the aperture, aspreviously estimated. However, the change in vacuolar contentof only 5.9 pmol mm^–2 µm^–1 is much too smallto account for the osmotic changes required, or to balance thecytoplasmic changes. It appears therefore that increasing KBroutside not only increases the cytoplasmic salt content, andthe Br flux at the tonoplast, but also stimulates the vacuolaraccumulation of some other solute.     

Photosynthetic parameters, pigment composition and respiration rates of the marine diatom Skeletonema costatum grown in continuous light and a 12:12 h light-dark cycle

Nutrient-sufficient cultures of a Trondheimsfjord (Norway) cloneof the marine centric diatom Skeleionema costatum (Grev.) Clevewere grown at 75 µmol m^–2 s^–1 and 15C at24 and 12 h daylength to study diurnal variations and the effectof daylength on pigment and chemical composition, photosyntheticparameters, dark respiration rates and scaled fluorescence excitationspectra (F), the latter used as estimates for the absorptionof energy available to Photosystem II. Specific growth rateswere 1.06 and 0.56 day^– in 24 and 12 h daylength, respectively,while dark respiration rates were generally 85% of the net growthrate. The Chla-normalized photosynthetic coefficients P^B_m anda^B were {small tilde}20–25% higher in continuous lightthan at 12 h daylength, while the Chla:C ratio was {small tilde}15%lower (0.051 versus 0.061 w:w). Thus, the carbon-normalizedcoefficients P^c_m and a^c were <11% lower at 24 h than at 12h daylength. The maximum quantum yield _max, the Chla:C ratioand F differed negligibly, as did the light saturation indexl_k, the N:C ratio and the ratios Chlc:Chla and Fucoxanthin:Chla. P^B_m and l_k did not exhibit diurnal variations at 24 hdaylength, and varied within 23% of the daily mean at 12 h daylength.Predictions of the daily gross photosynthetic rate based ondata for a given time of the day should thus not be >10%in error relative to an integrated value based on several datasets collected through 24 h. _max was 0.084–0.117 mol O₂(mol photons)^– for gross oxygen evolution. However, ifused in mathematical models for predicting the gross and netgrowth rates (i.e. the gross and net carbon turnover rates),‘practical’ values of 0.076 and 0.040 g-at C (molphotons)^–, respectively, should be employed. Correspondingly,values for a^B and P^B_m should be adjusted pro rata. ¹Present address: College of Marine Studies, Sjmannsveien 27,N-6008 lesund, Norway     

The Osmotic Pressure of Concentrated Solutions of Polyethylene Glycol 6000, and its Variation with Temperature

The vapour pressures of aqueous solutions of polyethylene glycol6000 have been measured (by equilibration with sucrose solutions)up to the saturation point at 25 °C (1.45 g g^–1 water).The reduced-osmotic-pressure (/c), when plotted versus concentration(c), rapidly and linearly increased up to a concentration ofabout 0.8 g g^–1 (crossing the similar plot for sucrose).Above this concentration, the reduced-osmotic-pressure rosemore slowly, but still more rapidly than sucrose. The maximumosmotic pressure achieved at saturation was nearly 18 MPa. Usingthe virial equation: /c= RT/M + RTA₂c, the calculated secondvirial coefficient (A₂) for the linear part is 4.5 x 10^–3mol g^–1, a value slightly greater than most literaturevalues at 25 °C. Data are cited showing that A₂ varies linearlyfrom 5–6 x 10^x3 at 0 °C, to zero at 80–90 °C     

Relation between Ultra-Violet Spectral Change and Nucleotide Binding of the Chloroplast Coupling Factor 1

Properties of the nucleotide binding sites on chloroplast couplingfactor 1 (CF₁) were studied by equilibrium dialysis and UV spectroscopy.From our direct binding studies, we identified at least fourkinds of ADP binding sites on CF₁; a barely dissociable ADPbinding site (site A), a slowly exchangeable high affinity sitewith dissociation constant (Kd) 0.021 µM (site B), anotherslowly exchangeable high affinity site with Kd 1.6 µM(site C) and several low affinity (Kd {small tilde}30 µM)sites. The Kd values for sites B and C of the other nucleotidestested were 0.5 µM and 16 µM (GDP), 8 µM and34 µM (CDP), 17 µM and 20 µM (UDP) and 1.4µM and 1.4 µM (PP₁). From a comparison of the observed UV spectral change and theamount of nucleotide bound to these sites, as calculated fromthe above Kd values, we concluded that the nucleotide bindingto site B or G induces UV spectral changes that are almost thesame in shape and magnitude. The estimated difference molarabsorption coefficient () was 3.4?10³M^–1_ADP cm^–1for ADP at 278 nm. Our conclusions were strengthened by thegood agreement between the observed spectra and the calculatedspectra (derived from the Kd and values of ADP and GDP) whenADP and GDP were added together to CF₁. The cause of the unusual behavior of GDP in the UV differencespectrum which was unexplained in our previous report was shownto be competition between the GDP added and previously boundADP at sites B and C; this distorted the real spectrum inducedby GDP. (Received October 3, 1983; Accepted February 13, 1984)