Presence of the Photoactive Reaction-Center Chlorophyll of PSI (P700) in Dark-Grown Cells of a Chlorophyll-Deficient Mutant of Chlorella kessleri

Compositions of pigments and polypeptides of pale green membranesthat had been isolated from dark-grown cells of a chlorophyll-deficientmutant of Chlorella kessleri were investigated. They containedChl a in a level corresponding to about 1% of that present inthe thylakoid membranes isolated from autotrophically grownwild-type cells and a trace amount of chlorophyllide a, butneither Chl b nor carotenoids. The polypeptide profile of themutant membranes was similar to that of membranes isolated fromwild-type cells that were grown in the dark. Neither the chlorophyll-bindingsubunits of PSI nor the apoproteins of LHCP were detected bySDS-PAGE and immunoblot analysis. However, the light-minus-darkdifference spectrum of the mutant membranes revealed the presenceof the reaction-center chlorophyll of PSI (P700) at a molarratio of 190 chlorophyll (Chl a plus Chlide a) per P700. P700was more stable than Chl a and Chlide a in the light so thatprolonged illumination led to a decline in the Chl/P700 ratioto 24. The initial rate of P700 photooxidation in the mutantmembranes was comparable to that in CP1 isolated from the dark-grownwild-type cells. Under illumination with strong light, the initialrate was decreased in parallel to the decrease in Chl/P700 ratio.The results suggest that most of Chi present in the mutant membranescan transfer excitation energy to P700. (Received March 13, 1998; Accepted August 7, 1998)     

Antenna Sizes of Photosystem I and Photosystem II in Chlorophyll b-Deficient Mutants of Rice. Evidence for an Antenna Function of Photosystem II Centers That are Inactive in Electron Transport

Light-harvesting capacities of photosystem I (PSI) and photosystemII (PSII) in a wild-type and three chlorophyll b-deficient mutantstrains of rice were determined by measuring the initial slopeof light-response curve of PSI and PSII electron transport andkinetics of light-induced redox changes of P-700 and Q_A, respectively.The light-harvesting capacity of PSI determined by the two methodswas only moderately reduced by chlorophyll b-deficiency. Analysisof the fluorescence induction that monitors time course of Q_Aphotoreduction showed that both relative abundance and antennasize of PSII_a decrease with increasing deficiency of chlorophyllb and there is only PSII_rß in chlorina 2 which totallylacks chlorophyll b. The numbers of antenna chlorophyll moleculesassociated with the mutant PSII centers were, therefore, threeto five times smaller than that of PSII_a in the wild type rice.Rates of PSII electron transport determined on the basis ofPSII centers in the three mutants were 60–70% of thatin the normal plant at all photon flux densities examined, indicatingthat substantial portions of the mutant PSII centers are inactivein electron transport. The initial slopes of light-responsecurves of PSII electron transport revealed that the functionalantenna sizes of the active populations of PSII centers in themutants correspond to about half that of PSII in the wild typerice. Thus, the numbers of chlorophyll molecules that serveas antenna of the oxygen-evolving PSII centers in the mutantsare significantly larger than those that are actually associatedwith each PSII center. It is proposed that the inactive PSIIserves as an antenna of the active PSII in the three chlorophyllb-deficient mutants of rice. In spite of the reduced antennasize of PSII, therefore, the total light-harvesting capacityof PSII approximately matches that of PSI in the mutants. (Received July 29, 1994; Accepted February 7, 1996)     

Reconstitution of the Photosystem I Secondary Quinone Acceptor (A1) in the P700-Fx Core Isolated from Synechococcus PCC 6301

By treating a F_A/F_B-depleted P700-F_x core from SynechococcusPCC 6301 with diethylether, most of the phylloquinone was removedwithout loss of P700. The 1 ms decay of P700⁺ in the originalcore was replaced by the 25 ns decay, which was interpretedas the backreaction occurring in a P700⁺     

NAD(P)H Dehydrogenase-Dependent Cyclic Electron Flow around Photosystem I in the Cyanobacterium Synechocystis PCC 6803: a Study of Dark-Starved Cells and Spheroplasts

Electron donation to P700⁺ through plastoquinone in the intersystemchain from both respiratory substrates and the photoreductantsin PSI has been shown to be mediated by the NAD(P)H-dehydrogenasecomplex (NDH) in Synechocystis PCC 6803 cells [Mi et al. (1992)Plant Cell Physiol. 33: 1233]. To confirm the participationof NDH in the cyclic electron flow around PSI, the redox kineticsof P700 and Chi fluorescence were analyzed in cells rendereddeficient in respiratory substrates by dark starvation and inspheroplasts. Dark-starved cells showed a high steady-state level of P700⁺under far-red (FR) illumination and the plastoquinone pool wasin a highly oxidized state. An NDH-defective mutant consistentlyshowed a high level of P700 oxidation under FR before and afterthe dark starvation. Donation of electrons either from exogenousNADPH or from photoreduced NADPH⁺ to the intersystem chain viaplastoquinone was demonstrated using spheroplasts from wild-typecells, but not those from the NDH-defective mutant, as monitoredby following changes in the kinetics of Chi fluorescence andthe redox state of P700. The electron flow to PSI via plastoquinone,mediated by NADPH, was sensitive to rotenone, Hg²⁺ ions and2-thenoyltrifluoroacetone, inhibitors of mitochondrial NDH andsuccinate dehydrogenase, but not to antimycin A. The pool sizeof electrons that can be donated to P700⁺ from the cytosol throughthe intersystem chain increased with increasing duration ofillumination time by actinic light and was sensitive to rotenonein both wild-type cells and spheroplasts, but no such resultswere obtained in the NDH-defective mutant of Synechocystis 6803.The results support our previous conclusion that NDH is a mediatorof both respiratory electron flow and cyclic electron flow aroundPSI to the intersystem chain in the cyanobacterium Synechocystis. (Received August 20, 1993; Accepted November 22, 1993)     

Crystallization of Photosystem I Complexes from the Thermophilic Cyanobacterium, Synechococcus vulcanus

PSI complexes were isolated from the thermophilic cyanobacterium,Synechococcus vulcanus, by mild detergent treatment of the thylakoidmembranes, purified by sucrose-density gradient centrifugation,and then crystallized. High resolution SDS-PAGE revealed thepresence of the product of the psaI gene in S. vulcanus PSIcomplexes and crystals. Crystals of the PSI complexes retainedall of the components of electron carriers and subunit polypeptides(including PsaX) known in cyanobacteria. (Received July 22, 1998; Accepted October 19, 1998)     

A Mutant of Arabidopsis thaliana That Defines a New Locus for Glycine Decarboxylation

A novel photorespiratory mutant of Arabidopsis thaliana, designatedgld2, was isolated based on a growth requirement for abnormallyhigh levels of atmospheric CO₂. Photosynthetic CO₂ fixationwas inhibited in the mutant following illumination in air butnot in atmosphere containing 2% O₂. Photosynthetic assimilationof ¹⁴CO₂ in an atmosphere containing 50% O₂ resulted in accumulationof 48% of the soluble label in glycine in the mutant comparedto 9% in the wild type. The rate of glycine decarboxylationby isolated mitochondria from the mutant was reduced to 6% ofthe wild type rate. In genetic crosses, the mutant complementedtwo previously described photorespiratory mutants of A. thalianathat accumulate glycine during photosynthesis in air due todefects in glycine decarboxylase (glyD, now designated gld1)and serine transhydroxymethylase (stm). Because glycine decarboxylaseis a complex of four enzymes, these results are consistent witha mutation in a glycine decarboxylase subunit other than thataffected in the gld1 mutant. The two gld loci were mapped tochromosomes 2 and 5, respectively. ³Present address: Department of Crop and Soil Sciences, MichiganState University, East Lansing, MI 48824, U.S.A. ⁴Present address: Department of Applied Bioscience, Facultyof Agriculture, Hokkaido University, Kita-Ku, Sapporo, 060 Japan ⁵Present address: Department of Biology, Carnegie Institutionof Washington, 290 Panama Street, Standford, CA 94305, U.S.A.     

Inhibition of photosystem I and photosystem II in chloroplasts by UV radiation

The effects of UV radiation on the low temperature fluorescenceand primary photochemistry of PSII and PSI of spinach chloroplastswere studied. Fluorescence induction curves at –196°Cwere measured at 695 nm for PSII fluorescence and at 730 nmfor PSI fluorescence to determine both the initial F_o and finalF_M levels. The primary photochemistry of PSII was measured asthe rate of photoreduction of C-550 at – 196°C, thatof PSI as the rate of photooxidation of P₇₀₀ at –196°C.The results were analyzed in terms of a model for the photosyntheticapparatus which accounts for the yields of fluorescence andprimary photochemistry. According to this analysis UV radiationincreases nonradiative decay processes at the reaction centerchlorophyll of PSII. However, the effect of UV radiation isnot uniform throughout the sample during irradiation so thataccount must be taken of the fraction of PSII reaction centerswhich have been irradiated at any given time. UV radiation alsoinactivates P700 and causes a slight increase in nonradiativedecay in the antenna chlorophyll of PSI. All fluorescence ofvariable yield, F_V = F_M–F_o, at 730 nm is due to energytransfer from PSII to PSI so that the sensitivity of Fv to UVradiation is the same at 730 and 695 nm. ¹Present address: Department of Biology, Faculty of Science,Toho University, Narashino, Chiba 275, Japan. ²Present address: Central Research Laboratories, Fuji PhotoFilm Co., Ltd., 105 Mizonuma, Asaka-Shi, Saitama 351, Japan. (Received September 10, 1975; )     

Photoinhibition and light-induced cyclic electron transport in ndhB(-) and psaE(-) mutants of Synechocystis sp. PCC 6803

The ndhB^– and psaE^– mutants of the cyanobacteriumSynechocystis sp. PCC 6803 are partly deficient in PSI-drivencyclic electron transport. We compared photoinhibition in thesemutants to the wild type to test the hypothesis that PSI cyclicelectron transport protects against photoinhibition. Photoinhibitorytreatment greatly accelerated PSI cyclic electron transportin the wild type and also in both the mutants. The psaE^–mutant showed rates of PSI cyclic electron transport similarto the wild type under all conditions tested. The ndhB^–mutant showed much lower rates of PSI cyclic electron transportthan the wild type following brief dark adaptation but exceededwild type rates after exposure to photoinhibitory light. Thewild type and both mutants showed similar rates of photoinhibitiondamage and photoinhibition repair at PSII. Photoinhibition atPSI was much slower than at PSII and was also similar betweenthe wild type and both mutants, despite the known instabilityof PSI in the psaE^– mutant. We conclude that photoinhibitorylight induces sufficient PSI-driven cyclic electron transportin both the ndhB^– and psaE^– mutants to fulfill anyrole that cyclic electron transport plays in protection againstphotoinhibition. ⁴ Corresponding author: E-mail, sherbert@uwyo.edu; Fax, +1-307-766-2851;Phone, +1-307-766-4353.     

New fava bean guard cell signaling mutant impaired in ABA-induced stomatal closure

We isolated a mutant from Vicia faba L. cv. House Ryousai. Itwilts easily under strong light and high temperature conditions,suggesting that its stomatal movement may be disturbed. We determinedresponses of mutant guard cells to some environmental stimuli.Mutant guard cells demonstrated an impaired ability to respondto ABA in 0.1 mM CaCl₂ and stomata did not close in thepresence of up to 1 mM ABA, whereas wild-type stomata closedwhen exposed to 10 µM ABA. Elevating external Ca²⁺caused a similar degree of stomatal closure in the wild typeand the mutant. A high concentration of CO₂ (700 µlliter^–1) induced stomatal closure in the wild type, butnot in the mutant. On the basis of these results, we proposethe working hypothesis that the mutation occurs in the regiondownstream of CO₂ and ABA sensing and in the region upstreamof Ca²⁺ elevation. The mutant is named fia (fava bean impairedin ABA-induced stomatal closure). ³ Corresponding author: E-mail, smoiwai{at}agri.kagoshima-u.ac.jp;Fax, +81-99-285-8556.     

Effect of Supra-High Irradiation on the Photosynthetic System of the Cyanophyte Synechocystis PCC 6714

Stability of thylakoid components under supra-high irradiancewas studied with the cyanophyte Synechocystis PCC 6714. Theactivity of overall photosynthesis was quickly inactivated (T_1/2=20min) under supra-high irradiance (300 W m^–2, white light).In parallel with the inactivation of photosynthesis, QA in PSII was also inactivated. Both inactivations were acceleratedby chloramphenicol (CAP) addition. The reactivation of PS IIrequired weak irradiation and was suppressed by CAP. However,PS I measured as P700 was very stable. The level of PS I measuredas P700 was not significantly reduced by the irradiation for12 h even in the presence of CAP while the level of Cyt b₅₅₉,component of PS II, was decreased markedly. The function ofPS I before and after supra-high irradiation with CAP was examinedby comparing sizes of P700 oxidation induced by a short flash,by a continuous light, and by determination of O₂-and ferredoxin-reduction.No difference was observed in PS I actions before and afterthe irradiation treatment. These results indicate that the PSI complex is very tolerant of supra-high irradiation. However,the cells grown under supra-high irradiance contained much fewerPS I and PS II complexes than Cyt b₆–f complexes. Theformer levels were reduced to a half to one fourth of thosebefore growth while the level of Cyt b₆–f complex wasnot reduced so much. A possible mechanism for changes in thylakoidcomposition under supra-high irradiation was discussed. (Received February 16, 1991; Accepted June 12, 1991)     

Absence of PsaC subunit allows assembly of photosystem I core but prevents the binding of PsaD and PsaE in Synechocystis sp. PCC6803

In photosystem I (PSI) of oxygenic photosynthetic organisms the psaC polypeptide, encoded by the psaC gene, provides the ligands for two [4Fe-4S] clusters, F_A and F_B. Unlike other cyanobacteria, two different psaC genes have been reported in the cyanobacterium Synechocystis 6803, one (copy 1) with a deduced amino acid sequence identical to that of tobacco and another (copy 2) with a deduced amino acid sequence similar to those reported for other cyanobacteria. Insertion of a gene encoding kanamycin resistance into copy 2 resulted in a photosynthesis-deficient strain, CDK25, lacking the PsaC, PsaD and PsaE polypeptides in isolated thylakoid membranes, while the PsaA/PsaB and PsaF subunits were found. Growth of the mutant cells was indistinguishable from that of wild-type cells under light-activated heterotrophic growth (LAHG). A reversible P700⁺ signal was detected by EPR spectroscopy in the isolated thylakoids during illumination at low temperature. Under these conditions, the EPR signals attributed to F_A and F_B were absent in the mutant strain, but a reversible F_x signal was present with broad resonances at g=2.079, 1.903, and 1.784. Addition of PsaC and PsaD proteins to the thylakoids gave rise to resonances at g=2.046, 1.936, 1.922, and 1.880; these values are characteristic of an interaction-type spectrum of F_A ^- and F_B ^-. In room-temperature optical spectroscopic analysis, addition of PsaC and PsaD to the thylakoids also restored a 30 ms kinetic transient which is characteristic of the P700⁺ [F_A/F_B]^- backreaction. Expression of copy 1 was not detected in cells grown under LAHG and under mixotrophic conditions. These results demonstrate that copy 2 encodes the PsaC polypeptide in PSI in Synechocystis 6803, while copy 1 is not involved in PSI; that the PsaC polypeptide is necessary for stable assembly of PsaD and PsaE into PSI complex in vivo; and that PsaC, PsaD and PsaE are not needed for assembly of PsaA-PsaB dimer and electron transport from P700 to F_x.     

Chlorophyll-Protein Complexes in Salix sp. 'Aquatica Gigantea' under Strong and Weak Light I. Spectral Characterization of the Chlorophyll-Protein Complexes

The distribution of chlorophyll in the chlorophyll-protein complexeswas studied in Salix sp. ‘aquatica gigantea’ grownunder high and low irradiance. The chlorophyll- containing bandsthat could be separated by SDS-polyacrylamide gel electrophoresisin strong and weak light numbered 9 to 13 and 9 to 11, respectively.In strong light the following bands were separated, in the orderof the highest to the lowest molecular weight: one to two chlorophylla/b-protein complexes, three to four chlorophyll a-containingbands similar to the P700-chlorophyll a-protein complex (CPIand its oligomers), three oligomers of the light-harvestingchlorophyll a/b-protein complex (LHCP^***, LHCP^**, LHCP^*), twochlorophyll a-protein complexes (CPa₂ and CPa₁), the light-harvestingchlorophyll a/b-protein complex (LHCP) and the protein freepigment (FP). In weak light the same chlorophyll-containingbands were separated with the exception that no high molecularweight chlorophyll a/b-protein complexes could be observed.In strong light the CPI complexes were the largest structuralcomponent of the chloroplast lamellae. In weak light the LHCPcomplexes together contributed the major proportion of the totalchlorophyll. The increase in the chlorophyll associated withthe LHCP complex was possibly caused by reorganization of thelamellar structure or by increased synthesis of the LHCP^** complex,which appeared to be a labile complex in weak light. (Received February 1, 1982; Accepted May 10, 1982)     

不同浓度NaCl和Na2CO3处理对菊芋幼苗光合及叶绿素荧光的影响

 以砂培菊芋(Helianthus tuberosus)幼苗作为试验材料,分别进行不同浓度NaCl (50、 100、150、200、250 mmol&;#8226;L^－1)和Na₂CO₃ (25、50、 75、100、125 mmol&;#8226;L^－1)胁迫处理,以1/2全营养液作为对照,处理7 d后研究NaCl和Na₂CO₃胁迫处理对菊芋幼苗叶片光合作用及叶绿素动力学 参数的影响。结果表明：1)在NaCl处理下,当浓度小于150 mmol&;#8226;L^－1时,增加了菊芋的叶绿素含量、净光合速率(Net photosynthetic rate, P_n)和气孔导度(Stomatal conductivity, G_s),对荧光参数PSⅡ的电子传递情况( F_m/F_o)、PSⅡ原初光能转换效率(F_v/F_m)、PSⅡ量子效率 (Actual quantum yield of PSⅡ under actinic irradiation,φPSⅡ)和光化学猝灭系数(Photochemical quenching coefficient, qP)和非 光化学猝灭系 数(Non-photochemical quenching coefficient, NPQ)没有显著影响,随着浓度的增加,各项生理指标与对照相比除了NPQ显著 增加,其余均显著降低;2)在Na₂CO₃胁迫处理下,随着Na₂CO₃浓度的增加,与对照相比菊芋幼苗叶绿素含量、P_n、G_s以及叶绿素a荧光诱导动力 学参数F_m/F_o、F_v/F_m、φPSⅡ和qP均显著降低,NPQ显著增加;3)就NaCl和Na₂CO₃相比而言,在相同Na⁺浓度情况下,处于Na₂CO₃胁迫下的菊芋 幼苗的叶绿素含量、P_n、G_s以及叶绿素a荧光诱导动力学参数F_m/F_o、F_v/F_m、φPSⅡ和qP下降幅度和NPQ的增加幅度均显著大于NaCl,这说明 NaCl和Na₂CO₃胁迫均对菊芋幼苗造成不同程度的伤害,但在相同Na⁺浓度情况下,Na₂CO₃的伤害程度大于NaCl。由此说明菊芋对盐的忍耐程度高 于碱。     

Isolation of the Photoactive Reaction Center Complex that Contains Three Types of Fe-S Centers and a Cytochrome c Subunit from the Green Sulfur Bacterium Chlorobium limicola f. thiosulfatophilum, Strain Larsen

The photoactive reaction center (RC) complex from the greensulfur bacterium Chlorobium limicola f. thiosulfatophilum, strainLarsen, was isolated after solubilization and ammonium sulfatefractionation followed by ion-exchange chromatography. The spectrumof the complex was almost identical with that of the similarRC complex isolated by Feiler et al. [(1992) Biochemistry 31:2608–2614] except for the presence of cytochrome c₅₅₁instead of c₅₅₃ in the latter study. A molecular ratio of BChla to P840 of the isolated RC complex was assayed to be 25–35.SDSPAGE analysis revealed that the isolated complex containedthree major polypeptides with apparent molecular masses of 68,41 and 21 kDa, respectively. The 21-kDa polypeptide was identifiedto be a heme-binding protein by staining the gel for peroxidaseactivity. The cytochrome c₅₅₁ was oxidized by flash light ina biphasic manner with half times of 90 and 390 µs, respectively,that coincided with the reduction half times of P840⁺. Threedistinct iron-sulfur centers assigned to F_A, F_B and F_x, respectively,from their g-values were detected by EPR spectroscopy at cryogenictemperature. These results suggest that the present preparationcontains a minimal functional unit of the RC of this bacterium,and that this complex appears to lie on a evolutionary linebetween RC's of purple bacteria and photosystem I. (Received August 18, 1992; Accepted October 28, 1992)     

Simultaneous CO2- and 16O2/18O2-gas exchange and fluorescence measurements indicate differences in light energy dissipation between the wild type and the phytochrome-deficient aurea mutant of tomato during water stress

The CO₂-, H₂O- and ¹⁶O₂/¹⁸O₂ isotopic-gas exchange and the fluorescencequenching by attached leaves of the wild-type and of the phytochrome-deficienttomato aurea mutant was compared in relation to water stressand the photon fluence rate. The chlorophyll content of aurealeaves was reduced and the ultra-structure of the chloroplastswas altered. Nevertheless, the maximum rate of net CO₂ uptakein air by the yellow-green leaves of the aurea mutant was similarto that by the dark-green wild-type leaves. However, less O₂was produced by the leaves of the aurea mutant than by leavesof the wild-type. This result indicates a reduced rate of photosyntheticelectron flux in aurea mutant leaves. No difference in bothphotochemical and non-photochemical fluorescence quenching wasfound between wild-type and aurea mutant leaves. Water stresswas correlated with a reversible decrease in the rates of bothnet CO₂ uptake and transpiration by wild-type and aurea mutantleaves. The rate of gross ¹⁶O₂ evolution by both wild-type andaurea mutant leaves was fairly unaffected by water stress. Thisresult shows that in both wild-type and aurea leaves, the photochemicalprocesses are highly resistant to water stress. The rate ofgross ¹⁸O₂ uptake by wild-type leaves increased during waterstress when the photon fluence rate was high. Under the sameconditions, the rate of gross ¹⁸O₂ uptake by ^aurea mutant leavesremained unchanged. The physiological significane of this differencewith respect to the (presumed) importance of oxygen reductionin photoprotection is discussed. Key words: Water stress, gas exchange, fluorescence quenching, Lycopersicon esculentum, mutant (tomato, aurea), energy dissipation     

Stoichiometries of Photosystem I and Photosystem II in Rice Mutants Differently Deficient in Chlorophyll b

Stoichiometries of photosystem I (PSI) and photosystem II (PSII)reaction centers in a cultivar of rice, Norin No. 8, and threechlorophyll b-deficient mutants derived from the cultivar wereinvestigated. Quantitation of PSI by photooxidation of P-700and chromatographic assay of vitamin K₁ showed that, on thebasis of chlorophyll, the mutants have higher concentrationsof PSI than the wildtype rice. Greater increases were observedin the PSII contents measured by photoreduction of Q_A, bindingof a radioactive herbicide and atomic absorption spectroscopyof Mn. Consequently, the PSII to PSI ratio increased from 1.1–1.3in the wild-type rice to 1.8 in chlorina 2, which contains noChl b, and to 2.0–3.3 in chlorina 11 and chlorina 14,which have chlorophyll a/b ratios of 9 and 13, respectively.Measurement of oxygen evolution with saturating single-turnoverflashes revealed that, whereas at most 20% of PSII centers areinactive in oxygen evolution in the wildtype rice, the non-functionalPSII centers amount to about 50% in the three mutant strains.The fluorescence induction kinetics was also analyzed to estimateproportions of the inactive PSII in the mutants. The data obtainedsuggest that plants have an ability to adjust the stoichiometryof the two photosystems and the functional organization of PSIIin response to the genetically induced deficiency of chlorophyllb. (Received July 29, 1994; Accepted February 7, 1996)     

Estimating chlorophyll a abundance from the 'phytoplankton colour' recorded by the Continuous Plankton Recorder survey: validation with simultaneous fluorometry

The green colour (measured with reference to standard colourcharts) of sections of the Continuous Plankton Recorder (CPR)filtering silk was compared with estimates of chlorophyll aconcentration derived from a fluorometer mounted on the CPRduring seven tows in the North Sea between February and May1991. After the green colour was assessed, the abundance ofphytoplankton cells on the filtering silks was quantified bymicroscope analysis. Data were collected for 115 10-nautical-milesamples over a total of seven cruises. For these 115 samples,there was only a weak (F_1.113 = 3.8, P = 0.05, r² = 0.03) positiverelationship between the colour of the filtering silk and thechlorophyll a concentration. However, when this comparison wasrestricted to four tows (68 10-nautical-mile samples) wherethe recorded phytoplankton cell abundance on the silks was verylow, there was a highly significant (F_1.66 /,69.1, P < 0.001,r² = 0.51) positive relationship between the silk colour andthe chlorophyll a concentration. By measuring the relative colourintensity of CPR standard colour categories and quantifyingthe individual variation in the assessment of colour, a theoreticalmodel was developed which pedicted that if the silks were colouredin direct proportion to the chlorophyll a concentration in thewater, then the expert r² for the relationship between silkcolour and chlorophyll a concentration would be 0.62. The greencolour recorded by the CPR survey was therefore identified asa quantitative index of chlorophyll a concentration, but onlywhen numbers of phytoplankton cells on the CPR silks are nothigh.     

Flag Leaf Photosynthesis of Triticum aestivum and Related Diploid and Tetraploid Species

Rates of net photosynthesis of the flag leaves of 15 genotypesof wheat and related species were measured throughout theirlife, using intact leaves on plants grown in the field. At thestage when rates were maximal, they were in general highestfor the diploid species, intermediate for the tetraploidspeciesand lowest for Triticum aestivum (means of 38, 32 and 28 mgCO₂ dm^–2 h^–1 respectively). Rates were stronglynegatively correlated with leaf area, leaf width and the meanplan area per mesophyll cell and positvely correlated with stomatalfrequency and number of veins per mm of leaf width. The differencesamong species in these attributes were mainly related to ploidylevel. It was not possible to determine the relative importanceof each anatomical feature, though the changes in stomatal frequencyhad only slight effects on stomatal conductance and the observeddifferences in rates of photosynthesis were much greater thanwould be expected from those in stomatal conductance alone. There was genetic variation in rates of light dependent oxygenevolution of isolated protoplasts and intact chloroplasts butno difference attributable to ploidy. The mean rate, 91 µmolO₂ mg^–1 chlorophyll h^–1, equivalent to 3.9 mg CO₂mg^-1chlorophyll h^-1 was considerably less than the rate of photosynthesisin comparable intact leaves, which was 7.2 mg CO₂ mg^–1chlorophyll h^–1. The total above-ground dry matter yields were least for thewild diploids T. urartu and T. thauodar and the wild tetraploidT. dicoccoides, but the other wild diploids produced as muchdry matter as the hexaploids. The prospects of exploiting differences in photosynthetic ratein the breeding of higher yielding varieties are discussed. Triticum aestivum L., wheat, Aegilops spp, photosynthesis, stomatal conductance, stomatal frequency, polyploidy     

Thylakoid Membrane-Bound, NADPH-Specific Pyridine Nucleotide Dehydrogenase Complex Mediates Cyclic Electron Transport in the Cyanobacterium Synechocystis sp. PCC 6803

The donation of electrons from NADPH to the intersystem chain,as monitored by an increase in Chl fluorescence, occurred inthe isolated thylakoid membranes of Synechocystis PCC 6803.The stimulation by NADPH of the methyl viologen-dependent photoreductionof dioxygen and of the reduction of P700⁺ after photooxidationin the presence of DCMU also confirmed the donation of electronsfrom NADPH to the electron carriers in the intersystem. Thesereactions were sensitive to rotenone, capsaicin, l-(2-thenoyl)-3,3,3-trifluoroacetoneand HgCl₂ but not to antimycin A or flavone. In contrast tothe thylakoid membranes from the wild type, those from a mutant,designated M55, in which a gene of a subunit of the pyridinenucleotide dehydrogenase complex (NDH) had been inactivated,did not show evidence of such reactions. These results supportour previous hypothesis that the transport of electrons fromNADPH to the intersystem chain is mediated by NDH [Mi et al.(1994) Plant Cell Physiol. 35: 163] and indicate the bindingof an NADPH-specific NDH to the thylakoid membranes. The Chlfluorescence was quenched transiently by addition of ferredoxinand NADP₊ to the thylakoid membranes but showed a subsequentincrease. This result suggests the reduction of plastoquinoneby the photoreduced NADP₊ and initiation of the NADPH-mediatedcyclic flow of electrons around PSI. Furthermore, a similarresponse of Chl fluorescence was observed upon the additionof ferredoxin only, demonstrating the ferredoxin-dependent cyclicflow of electrons. Both pathways of cyclic electron transportwere inhibited by rotenone, and were not detected in the NDH-defectedthylakoid membranes from M55, indicating the participation ofthe NDH complex. These results confirm that, in Synechocystis,the thylakoid-bound NDH complex mediates the ferredoxin-dependentcyclic electron flow, as well as the NADPH-dependent cyclicelectron flow. (Received November 24, 1994; Accepted March 16, 1995)     

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)