Chl Fluorescence Yield Research Articles

Salt Stress Induces Paramylon Accumulation and Fine-Tuning of the Macro-Organization of Thylakoid Membranes in Euglena gracilis Cells.

The effects of salt stress condition on the growth, morphology, photosynthetic performance, and paramylon content were examined in the mixotrophic, unicellular, flagellate Euglena gracilis. We found that salt stress negatively influenced cell growth, accompanied by a decrease in chlorophyll (Chl) content. Circular dichroism (CD) spectroscopy revealed the changes in the macro-organization of pigment-protein complexes due to salt treatment, while the small-angle neutron scattering (SANS) investigations suggested a reduction in the thylakoid stacking, an effect confirmed by the transmission electron microscopy (TEM). At the same time, the analysis of the thylakoid membrane complexes using native-polyacrylamide gel electrophoresis (PAGE) revealed no significant change in the composition of supercomplexes of the photosynthetic apparatus. Salt stress did not substantially affect the photosynthetic activity, as reflected by the fact that Chl fluorescence yield, electron transport rate (ETR), and energy transfer between the photosystems did not change considerably in the salt-grown cells. We have observed notable increases in the carotenoid-to-Chl ratio and the accumulation of paramylon in the salt-treated cells. We propose that the accumulation of storage polysaccharides and changes in the pigment composition and thylakoid membrane organization help the adaptation of E. gracilis cells to salt stress and contribute to the maintenance of cellular processes under stress conditions.

Modeling leaf CO2 assimilation and Photosystem II photochemistry from chlorophyll fluorescence and the photochemical reflectance index.

We present a simple model to assess the quantum yield of photochemistry (ΦP ) and CO2 assimilation rate from two parameters that are detectable by remote sensing: chlorophyll (chl) fluorescence and the photochemical reflectance index (PRI). ΦP is expressed as a simple function of the chl fluorescence yield (ΦF ) and nonphotochemical quenching (NPQ): ΦP =1-bΦF (1+NPQ). Because NPQ is known to be related with PRI, ΦP can be remotely assessed from solar-induced fluorescence and the PRI. The CO2 assimilation rate can be assessed from the estimated ΦP value with either the maximum carboxylation rate (Vcmax ), the intercellular CO2 concentration (Ci ), or parameters of the stomatal conductance model. The model was applied to experimental data obtained for Chenopodium album leaves under various environmental conditions and was able to successfully predict ΦF values and the CO2 assimilation rate. The present model will improve the accuracy of assessments of gas exchange rates and primary productivity by remote sensing.

Chlorophyll fluorescence, yield and yield components of bread wheat affected by phosphate bio-fertilizer, zinc and boron under late-season heat stress

We examined effects of late-season heat stress (L-SHS) on chlorophyll (Chl) fluorescence parameters and yield of bread wheat as well as roles of phosphate bio-fertilizer (PB-F) and Zn and B to compensate for the likely effects of heat stress. Factors were planting date (21 November and 5 January to coincide with grain filling to L-SHS) as the main factor, no inoculation (control) and inoculation of the seeds with PB-F as the sub-factor, and foliar application of water (control), Zn, B, and Zn + B as 3 L ha-1 as sub-sub factor. Results revealed that L-SHS reduced maximal quantum yield of PSII photochemistry, effective quantum yield of PSII photochemistry, efficiency of PSII in the light-adapted state, and the grain yield. Moreover, L-SHS increased the nonphotochemical quenching. The PB-F mitigated the effects of L-SHS on Chl fluorescence, yield, and yield components. Among nutrients, the combined Zn + B was more effective in reducing the effects of L-SHS than that of Zn and B alone. Nevertheless, there was an interaction between foliar nutrients application and PB-F, suggesting that Zn application alone had a profound influence on improving Chl fluorescence parameters and increased yield in combination with PB-F.

Substituting Fe for two of the four Mn ions in photosystem II-effects on water-oxidation.

We have investigated the interaction of Fe(II) cations with Ca-depleted PSII membranes (PSII[-Ca,4Mn]) in the dark and found that Fe(II) incubation removes 2 of 4Mn ions from the tetranuclear Mn cluster of the photosynthetic O2-evolving complex (OEC). The reduction of Mn ions in PSII(-Ca,4Mn) by Fe(II) and the concomitant release of two Mn(II) cations is accompanied by the binding of newly generated Fe(III) in at least one vacated Mn site. Flash-induced chlorophyll (Chl) fluorescence yield measurements of this new 2Mn/nFe cluster (PSII[-Ca,2Mn,nFe]) show that charge recombination in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) occurs between Qa (-) and the remaining Mn/Fe cluster (but not YZ (●)) in the OEC, and extraction of 2Mn occurs uniformly in all PSII complexes. No O2 evolution is observed, but the heteronuclear metal cluster in PSII(-Ca,2Mn,nFe) samples is still able to supply electrons for reduction of the exogenous electron acceptor, 2,6-dichlorophrenolindophenol, by photooxidizing water and producing H2O2 in the absence of an exogenous donor as seen previously with PSII(-Ca,4Mn). Selective extraction of Mn or Fe cations from the 2Mn/nFe heteronuclear cluster demonstrates that the high-affinity Mn-binding site is occupied by one of the iron cations. It is notable that partial water-oxidation function still occurs when only two Mn cations are present in the PSII OEC.

Oxidation of plastohydroquinone by photosystem II and by dioxygen in leaves

In sunflower leaves linear electron flow LEF=4O2 evolution rate was measured at 20ppm O2 in N2. PSII charge separation rate CSRII=aII∙PAD∙(Fm−F)/Fm, where aII is excitation partitioning to PSII, PAD is photon absorption density, Fm and F are maximum and actual fluorescence yields. Under 630nm LED+720nm far-red light (FRL), LEF was equal to CSRII with aII=0.51 to 0.58. After FRL was turned off, plastoquinol (PQH2) accumulated, but LEF decreased more than accountable by F increase, indicating PQH2-oxidizing cyclic electron flow in PSII (CEFII). CEFII was faster under conditions requiring more ATP, consistent with CEFII being coupled with proton translocation. We propose that PQH2 bound to the QC site is oxidized, one e− moving to P680+, the other e− to Cyt b559. From Cyt b559 the e− reduces QB− at the QB site, forming PQH2. About 10–15% electrons may cycle, causing misses in the period-4 flash O2 evolution and lower quantum yield of photosynthesis under stress. We also measured concentration dependence of PQH2 oxidation by dioxygen, as indicated by post-illumination decrease of Chl fluorescence yield. After light was turned off, F rapidly decreased from Fm to 0.2 Fv, but further decrease to F0 was slow and O2 concentration dependent. The rate constant of PQH2 oxidation, determined from this slow phase, was 0.054s−1 at 270μM (21%) O2, decreasing with Km(O2) of 60μM (4.6%) O2. This eliminates the interference of O2 in the measurements of CEFII.

Structural and Functional Analysis of the Antiparallel Strands in the Lumenal Loop of the Major Light-harvesting Chlorophyll a/b Complex of Photosystem II (LHCIIb) by Site-directed Mutagenesis

The light-harvesting chlorophyll a/b-binding protein of photosystem II (LHCIIb) fulfills multiple functions, such as light harvesting and energy dissipation under different illuminations. The crystal structure of LHCIIb at the near atomic resolution reveals an antiparallel strands structure in the lumenal loop between the transmembrane helices B/C. To study the structural and functional significances of this structure, three amino acids (Val-119, His-120, and Ser-123) in this region have been exchanged to Phe, Leu, and Gly, respectively, and the influence of the mutagenesis on the structure and function of LHCIIb has been investigated. The results are as follows. 1) Circular dichroism spectra of the mutations reveals that the antiparallel strands in the lumenal region are very important for adjusting pigment conformation in the neoxanthin domain of LHCIIb. Although the mutagenesis causes only a slight loss of the Neo binding in the complexes (V119F, 0.09; S123G, 0.19; and H120L, 0.27), it imparts remarkable changes to the pigment conformation. 2) Substituting Ser-123 with Gly results in a higher susceptibility to photodamage, an increased tendency to aggregate, and enhanced fluorescence quenching induced by the medium acidification. These results demonstrate that this antiparallel strands domain plays an important role in regulating the pigment conformation and in adjusting the aggregation and the fluorescence yield of LHCIIb.

A rapid, whole‐tissue determination of the functional fraction of PSII after photoinhibition of leaves based on flash‐induced P700 redox kinetics

Assaying the number of functional PSII complexes by the oxygen yield from leaf tissue per saturating, single-turnover flash, assuming that each functional PSII evolves one oxygen molecule after four flashes, is one of the most direct methods but time-consuming. The ratio of variable to maximum Chl fluorescence yield (F(v)/F(m)) in leaves can be correlated with the oxygen yield per flash during a progressive loss of PSII activity associated with high-light stress and is rapid and non-intrusive, but suffers from being representative of chloroplasts near the measured leaf surface; consequently, the exact correlation depends on the internal leaf structure and on which leaf surface is being measured. Our results show that the average F(v)/F(m) of the adaxial and abaxial surfaces has a reasonable linear correlation with the oxygen yield per flash after varied extents of photoinactivation of PSII. However, we obtained an even better linear correlation between (1) the integrated, transient electron flow (Sigma) to P700+, the dimeric Chl cation in PSI, after superimposing a single-turnover flash on steady background far-red light and (2) the relative oxygen yield per flash. Leaves of C3 and C4 plants, woody and herbaceous species, wild-type and a Chl-b-less mutant, and monocot and dicot plants gave a single straight line, which seems to be a universal relation for predicting the relative oxygen yield per flash from Sigma. Measurement of Sigma is non-intrusive, representative of the whole leaf tissue, rapid and applicable to attached leaves; it may even be applicable in the field.

Photosynthesis and protective mechanisms during ageing in transgenic tobacco leaves with over-expressed cytokinin oxidase/dehydrogenase and thus lowered cytokinin content

The content of cytokinins (CKs), the plant inhibitors of the final phase of plant development, senescence, is effectively controlled by irreversible degradation catalysed by cytokinin oxidase/dehydrogenase (CKX). In transgenic tobacco, denoted as AtCKX, with over-expressed CKX causing lowered CK content, we investigated changes in the time courses of chlorophyll (Chl) and xanthophyll (violaxanthin, antheraxanthin, zeaxanthin, neoxanthin, and lutein) contents. We also determined parameters of slow Chl fluorescence kinetics such as minimum Chl fluorescence yield in the darkadapted state F0, maximum quantum yield of PS2 photochemistry (Fv/Fm), maximum ratio of quantum yields of photochemical and concurrent non-photochemical processes in photosystem 2 (PS2), Fv/F0, non-photochemical quenching (NPQ), and effective quantum yield of photochemical energy conversion in PS2 (Φ2). We used three different developmental leaf stages, old, mature, and young, and compared this with time courses of these characteristics in leaves with natural CK levels. The parameters Fv/Fm, Fv/F0, and Φ2 were unchanged during ageing in AtCKX plants in contrast to control ones where a significant decrease in old leaves was found. In control plants F0 increased during ageing, but in the oldest leaf a considerable decrease was observed. This could indicate progressive damage to PS2 reaction centres and then detachment and rapid degradation of Chl. This is in agreement with time course of Chl content. NPQ decreased with age and was similar in both plant types. We observed a decline of xanthophyll contents in the oldest leaves in both plant types, but the contents were enhanced in AtCKX compared to control plants, especially of neoxanthin. The higher xanthophyll contents in the transgenic plants contribute to a better photoprotection and the fluorescence parameters indicated that photosynthetic apparatus was in better condition compared to control and it consequently postponed the onset of leaf senescence.

Chlorophyll fluorescence imaging of photosynthetic activity with the flash-lamp fluorescence imaging system

A flash-lamp chlorophyll (Chl) fluorescence imaging system (FL-FIS) is described that allows to screen and image the photosynthetic activity of several thousand leaf points (pixels) of intact leaves in a non-destructive way within a few seconds. This includes also the registration of several thousand leaf point images of the four natural fluorescence bands of plants in the blue (440 nm) and green (520 nm) regions as well as the red (near 690 nm) and far-red (near 740 nm) Chl fluorescence. The latest components of this Karlsruhe FL-FIS are presented as well as its advantage as compared to the classical single leaf point measurements where only the fluorescence information of one leaf point is sensed per each measurement. Moreover, using the conventional He-Ne-laser induced two-wavelengths Chl fluorometer LITWaF, we demonstrated that the photosynthetic activity of leaves can be determined measuring the Chl fluorescence decrease ratio, RFd (defined as Chl fluorescence decrease Fd from maximum to steady state fluorescence Fs:Fd/Fs), that is determined by the Chl fluorescence induction kinetics (Kautsky effect). The height of the values of the Chl fluorescence decrease ratio RFd is linearly correlated to the net photosynthetic CO2 fixation rate PN as is indicated here for sun and shade leaves of various trees that considerably differ in their PN. Imaging the RFd-ratio of intact leaves permitted the detection of considerable gradients in photosynthetic capacity across the leaf area as well as the spatial heterogeneity and patchiness of photosynthetic quantum conversion within the control leaf and the stressed plants. The higher photosynthetic capacity of sun versus shade leaves was screened by Chl fluorescence imaging. Profile analysis of fluoresence signals (along a line across the leaf area) and histograms (the signal frequency distribution of the fluorescence information of all measured leaf pixels) of Chl fluorescence yield and Chl fluorescence ratios allow, with a high statistical significance, the quantification of the differences in photosynthetic activity between various areas of the leaf as well as between control leaves and water stressed leaves. The progressive uptake and transfer of the herbicide diuron via the petiole into the leaf of an intact plant and the concomitant loss of photosynthetic quantum conversion was followed with high precision by imaging the increase of the red Chl fluorescence F690. Differences in the availability and absorption of soil nitrogen of crop plants can be documented via this flash-lamp fluorescence imaging technique by imaging the blue/red ratio image F440/F690, whereas differences in Chl content are detected by collecting images of the fluorescence ratio red/far-red, F690/F740.

Heat stress induces an inhibition of excitation energy transfer from phycobilisomes to photosystem II but not to photosystem I in a cyanobacterium Spirulina platensis

The effects of high temperature (30–52.5 °C) on excitation energy transfer from phycobilisomes (PBS) to photosystem I (PSI) and photosystem II (PSII) in a cyanobacterium Spirulina platensis grown at 30 °C were studied by measuring 77 K chlorophyll (Chl) fluorescence emission spectra. Heat stress had a significant effect on 77 K Chl fluorescence emission spectra excited either at 436 or 580 nm. In order to reveal what parts of the photosynthetic apparatus were responsible for the changes in the related Chl fluorescence emission peaks, we fitted the emission spectra by Gaussian components according to the assignments of emission bands to different components of the photosynthetic apparatus. The 643 and 664 nm emissions originate from C-phycocyanin (CPC) and allophycocyanin (APC), respectively. The 685 and 695 nm emissions originate mainly from the core antenna complexes of PSII, CP43 and CP47, respectively. The 725 and 751 nm band is most effectively produced by PSI. There was no significant change in F725 and F751 during heat stress, suggesting that heat stress had no effects on excitation energy transfer from PBS to PSI. On the other hand, heat stress induced an increase in the ratio of Chl fluorescence yield of PBS to PSII, indicating that heat stress inhibits excitation energy transfer from PBS to PSII. However, this inhibition was not associated with an inhibition of excitation energy transfer from CPC to APC since no significant changes in F643 occurred at high temperatures. A dramatic enhancement of F664 occurring at 52.5 °C indicates that excitation energy transfer from APC to the PSII core complexes is suppressed at this temperature, possibly due to the structural changes within the PBS core but not to a detachment of PBS from PSII, resulting in an inhibition of excitation energy transfer from APC to PSII core complexes (CP47 + CP43). A decrease in F685 and F695 in heat-stressed cells with excitation at 436 nm seems to suggest that heat stress did not inhibit excitation energy transfer from the Chl a binding proteins CP47 and CP43 to the PSII reaction center and the decreased Chl fluorescence yields from CP43 and CP47 could be explained by the inhibition of the energy transfer from APC to PSII core complexes (CP47 + CP43).

Differential changes in degradation of chlorophyll–protein complexes of photosystem I and photosystem II during flag leaf senescence of rice

The stability of chlorophyll–protein complexes of photosystem I (PSI) and photosystem II (PSII) was investigated by chlorophyll (Chl) fluorescence spectroscopy, absorption spectra and native green gel separation system during flag leaf senescence of two rice varieties (IIyou 129 and Shanyou 63) grown under outdoor conditions. During leaf senescence, photosynthetic CO 2 assimilation rate, carboxylase activity of Rubisco, chlorophyll and carotenoids contents, and the chlorophyll a/ b ratio decreased significantly. The 77 K Chl fluorescence emission spectra of thylakoid membranes from mature leaves had two peaks at around 685 and 735 nm emitting mainly from PSII and PSI, respectively. The total Chl fluorescence yields of PSI and PSII decreased significantly with senescence progressing. However, the decrease in the Chl fluorescence yield of PSI was greater than in the yield of PSII, suggesting that the rate of degradation in chlorophyll–protein complexes of PSI was greater than in chlorophyll–protein complexes of PSII. The fluorescence yields for all chlorophyll–protein complexes decreased significantly with leaf senescence in two rice varieties but the extents of their decrease were significantly different. The greatest decrease in the Chl fluorescence yield was in PSI core, followed by LHCI, CP47, CP43, and LHCII. These results indicate that the rate of degradation for each chlorophyll–protein complex was different and the order for the stability of chlorophyll–protein complexes during leaf senescence was: LHCII > CP43 > CP47 > LHCI > PSI core, which was partly supported by the green gel electrophoresis of the chlorophyll–protein complexes.

Functional asymmetry of photosystem II D1 and D2 peripheral chlorophyll mutants of Chlamydomonas reinhardtii.

The peripheral accessory chlorophylls (Chls) of the photosystem II (PSII) reaction center (RC) are coordinated by a pair of symmetry-related histidine residues (D1-H118 and D2-H117). These Chls participate in energy transfer from the proximal antennae complexes (CP43 and CP47) to the RC core chromophores. In addition, one or both of the peripheral Chls are redox-active and participate in a low-quantum-yield electron transfer cycle around PSII. We demonstrate that conservative mutations of the D2-H117 residue result in decreased Chl fluorescence quenching efficiency attributed to reduced accumulation of the peripheral accessory Chl cation, Chl(Z)(+). In contrast, identical symmetry-related mutations at residue D1-H118 had no effect on Chl fluorescence yield or quenching kinetics. Mutagenesis of the D2-H117 residue also altered the line width of the Chl(Z)(+) EPR signal, but the line shape of the D1-H118Q mutant remained unchanged. The D1-H118 and D2-H117 mutations also altered energy transfer properties in PSII RCs. Unlike wild type or the D1-H118Q mutant, D2-H117N RCs exhibited a reduced CD doublet in the red region of Chl absorbance band, indicative of reduced energetic coupling between P680 and the peripheral accessory Chl. In addition, transient absorption measurements of D2-H117N RCs, excited on the blue side of the Chl absorbance band, exhibited a ( approximately 400 fs) pheophytin Q(X) band bleach lifetime component not seen in wild-type or D1-H118Q RCs. The origin of this component may be related to delayed fast-energy equilibration of the excited state between the core pigments of this mutant.

パルス振幅変調（ＰＡＭ）クロロフィル蛍光測定法による野菜の高温ストレス耐性の検定

Pulse amplitude modulation (PAM) chlorophyll (Chl) fluorescence yield measurement was evaluated for estimating the high-temperature stress tolerance of vegetable crops. For this purpose, twenty two kinds of vegetable crops comprising of 17 taxonomic species were grown in controlled environments at the following three temperature regimes : (1) the air temperature during the 14-hr photo period was maintained at 20°C ; (2) it was raised stepwise at 5°C hr-1 from 20°C to 35°C, and then immediately decreased stepwise to 20°C, and (3) it was changed similarly to (2) except that the duration at 35°C was prolonged to 3 hr. At night the ambient air was cooled to 10°C in all three pretreatments. Following the pretreatments for 48 days, photosystem (PS) II Chl. fluorescence yields of leaf sections were determined by PAM before and during exposure of leaf sections to 45°C for up to 5 hr. Based on the heat sensitivity of PS II fluorescence yields, the vegetable crops tested were grouped into the following three classes : (A) crops with heat-tolerant PS II, (B) those with PS II sensitive to heat but capable of high-temperature acclimation, and (C) those with PS II sensitive to heat and incapable of high-temperature acclimation. These groupings coincide well with those based on the field performance of the crops at high temperatures. It is thus concluded that the Chl fluorescence yield measurement is a simple and reliable method for estimating the high-temperature stress tolerance of vegetable crops.

Changes in Chlorophyll a Fluorescence, Lipid Peroxidation, and Detoxificant System in Potato Plants Grown under Filtered and Non-Filtered Air in Open-Top Chambers

Its high oxidant capacity and ability to generate reactive oxygen species cause ozone toxicity. We studied the effect of ambient ozone on chlorophyll (Chl) a fluorescence, antioxidant enzymes, ascorbate contents, and lipid peroxidation in potatoes grown in open-top chambers in the field. In plants grown in non-filtered air (NFA), the development of non-photochemical quenching brought about a decrease in photosystem 2 (PS2) photochemical efficiency. Also the ability of PS2 to reduce the primary acceptor QA was lower than in charcoal-filtered, ozone-free air (CFA). Changes in Chl fluorescence yield were associated with changes in the thylakoid membrane. Ozone altered chloroplast membrane properties, as indicated by an increase in membrane lipid peroxidation in FNA-leaves compared to CFA plants. The ascorbate pool and activities of antioxidant enzymes were used for an indication of the detoxification system state in NFA and CFA leaves, whereby ozone affects the ascorbate concentration and decreases the antioxidant enzymes activities. The capacity of both detoxifying systems together was not high enough to protect potato plants against ambient ozone concentrations which reduced the photosynthetic yield in this potato cultivar.

Measurement of Differences in Red Chlorophyll Fluorescence and Photosynthetic Activity between Sun and Shade Leaves by Fluorescence Imaging

With a flash-lamp chlorophyll (Chl) fluorescence imaging system (FL-FIS) the photosynthetic activity of several thousand image points of intact shade and sun leaves of beech were screened in a non-destructive way within a few seconds. The photosynthetic activity was determined via imaging the Chl fluorescence at maximum Fp and steady state fluorescence Fs of the induction kinetics (Kautsky effect) and by a subsequent determination of the images of the fluorescence decrease ratio RFd and the ratio Fp/Fs. Both fluorescence ratios are linearly correlated to the photosynthetic CO2 fixation rates. This imaging method permitted to detect the gradients in photosynthetic capacity and the patchiness of photosynthetic quantum conversion across the leaf. Sun leaves of beech showed a higher photosynthetic capacity and differential pigment ratios (Chl a/b and Chls/carotenoids) than shade leaves. Profile analysis and histogram of the Chl fluorescence yield and the Chl fluorescence ratios allow to quantify the differences in photosynthetic activity between different leaf parts and between sun and shade leaves with a high statistical significance.

Photosynthetic characteristics of marine phytoplankton from pump-during-probe fluorometry of individual cells at sea.

Active fluorescence techniques are becoming commonly used to monitor the state of the photosynthetic apparatus in natural populations of phytoplankton, but at present these are bulk water measurements that average all the fluorescent material in each sample. Here we describe two instruments that combine individual-cell "pump-during-probe" (PDP) measurements of chlorophyll (Chl) fluorescence induction, on the time scale of 30 to 100 micros, with flow cytometric or visual characterization of each cell. In the PDP flow cytometer, we measure Chl fluorescence yield as a function of time during a 150 micros excitation flash provided by an argon ion laser; each particle is subsequently classified as in a conventional flow cytometer. In the PDP microfluorometer, individual cells in a sample chamber are visually identified, and fluorescence excitation is provided by a blue light-emitting diode that can be configured to provide a saturating flash and also a subsequent series of short flashlets. This sequence allows both saturation and relaxation kinetics to be monitored. Phytoplankton from natural samples and on-deck iron-enrichment incubation experiments in the Southern Ocean were examined with each PDP instrument, providing estimates of the potential quantum yield of photochemistry and the functional absorption cross section for photosystem 2, for either individuals (for cells larger than a few micrometers) or populations (for smaller cells). Results from initial field applications indicate that single-cell PDP measurements can be a powerful tool for investigating the nutritional state of phytoplankton cells and the regulation of phytoplankton growth in the sea.

Changes of Chlorophyll(ide) Fluorescence Yield Induced by a Short Light Pulse as a Probe to Monitor the Early Steps of Etioplast Phototransformation in Dark‐Grown Leaves

ABSTRACTThe fluorescence yield of chlorophyll(ide) (Chl[ide]) excited by weak modulated light was recorded at room temperature during a 2 h period after a short actinic light pulse that transformed all photoactive protochlorophyllide in dark‐grown barley leaves. A typical pattern of fluorescence yield variations was found whatever the age of the leaf but with age‐dependent changes in rates. Its successive phases were related to the Chl(ide) spectral shifts observed in low‐temperature emission spectra. The fluorescence yield started at a high level and strongly declined during the formation of Chlide695 from Chlide668 within a few seconds. It increased to a transient maximum during the Shibata shift (15–25 min) that resulted in Chl(ide)682. A final, slow decrease to a steady state occurred during the final red shift to Chl685. Pretreatments with δ‐aminolevulinic acid, chloramphenicol or 1, 10‐phenanthroline resulted in correlated modifications of Chl(ide) fluorescence yield transients and shifts of the low‐temperature Chl(ide) emission band. The complex response of the final decrease phase of the fluorescence yield to these compounds suggests that it results both from the assembly of photosynthetic Chl proteins and from the reorganization of the etioplast membrane system. From these results it is concluded that continuous recordings of Chl(ide) fluorescence yield after a short light pulse represent a useful tool to monitor the kinetics of pigment–protein organization and primary thylakoid assembly triggered by Pchlide photoreduction.

Changes of Chlorophyll(ide) Fluorescence Yield Induced by a Short Light Pulse as a Probe to Monitor the Early Steps of Etioplast Phototransformation in Dark-Grown Leaves

The fluorescence yield of chlorophyll(ide) (Chl[ide]) excited by weak modulated light was recorded at room temperature during a 2 h period after a short actinic light pulse that transformed all photoactive protochlorophyllide in dark-grown barley leaves. A typical pattern of fluorescence yield variations was found whatever the age of the leaf but with age-dependent changes in rates. Its successive phases were related to the Chl(ide) spectral shifts observed in low-temperature emission spectra. The fluorescence yield started at a high level and strongly declined during the formation of Chlide695 from Chlide668 within a few seconds. It increased to a transient maximum during the Shibata shift (15–25 min) that resulted in Chl(ide)682. A final, slow decrease to a steady state occurred during the final red shift to Chl685. Pretreatments with δ-aminolevulinic acid, chloramphenicol or 1, 10-phenanthroline resulted in correlated modifications of Chl(ide) fluorescence yield transients and shifts of the low-temperature Chl(ide) emission band. The complex response of the final decrease phase of the fluorescence yield to these compounds suggests that it results both from the assembly of photosynthetic Chl proteins and from the reorganization of the etioplast membrane system. From these results it is concluded that continuous recordings of Chl(ide) fluorescence yield after a short light pulse represent a useful tool to monitor the kinetics of pigment–protein organization and primary thylakoid assembly triggered by Pchlide photoreduction.

Nuclear, Virescent Mutants of Zea mays L. with High Levels of Chlorophyll (a/b) Light-Harvesting Complex during Thylakoid Assembly

We have found nuclear, recessive mutants in Zea mays L. where assembly of the major chlorophyll (a/b) light-harvesting complex (LHC) was not delayed relative to most other thylakoid protein complexes during thylakoid biogenesis. This contrasts with the normal development of maize chloroplasts (NR Baker, R Leech 1977 Plant Physiol 60: 640-644). All four mutants examined were allelic and virescent, and displayed visibly higher yields of leaf Chl fluorescence during greening. Fully greened mutants had normal leaf Chl fluorescence yield and normal levels of LHC, and grew to maturity under field conditions. Therefore, delayed LHC assembly is not an obligate feature of thylakoid differentiation.Assigning the molecular basis for the mutation should provide information concerning reguation of LHC assembly. Several possibilities are discussed. The pleiotropic mutant phenotype is not attributable to defects in thylakoid glycerolipid synthesis. Thylakoids isolated from greening mutant leaf sections had elevated glycerolipid/Chl ratios. In addition, both the molar distribution and acyl composition of four major glycerolipids were normal for developing mutant thylakoids.

Thermoluminescence and temperature effects on delayed light emission (corrected for changes in quantum yield of fluorescence) in DCMU-treated algae.

Abstract— The simultaneous measurements of delayed light emission (DLE) and chlorophyll (Chl) fluorescence yield in DCMU§ treated Chlorella were made in the time range of 1 to 10 sec at various temperatures from 0 to 50°C. Similar measurements were made for DCMU treated thermophilic strain of Synechococcus in the temperature range of 0 to 75°C. Using the basic assumption that DLE is produced by the back reaction of primary photoproducts of system II, and that two such reactions are required for it, a linear relationship between J‐1/2 (where J is energy per unit time available for DLE) and time after illumination was derived. This second‐order relationship was confirmed experimentally at several temperatures (2°, 5°, 10° and 15°C). From these analyses, reaction rate decay constants, at specific temperatures, were calculated. An Arrhenius plot was made for these calculated rate constants. Its slope (8–10 kcal/mole) agreed well with previous reports; however, it had a region of zero slope which occurred at the physiological temperature of the organisms used. Thermoluminescence or temperature jump delayed light emission (TDLE) was measured using various temperature conditions and it was found that not only the magnitude of the temperature jump (ΔT), but the initial and final temperatures of the sample were important. For example, a temperature jump of 8°C from 2 to 10°C gave much higher TDLE than from 12 to 20°C. Many properties e.g., magnitude, temperature dependence and time independence of TDLE could be explained by the DLE decay data (corrected for changes in fluorescence yield) and the kinetic analysis. It is suggested that, in addition to the back reaction of Z+ (the primary oxidized photoproduct of system II) with Q‐ (the primary reduced photoproduct of system II), a reducing entity, beyond the sites of DCMU and antimycin a action, is somehow involved in the production of slow DLE.