Chlorina F2 Research Articles

Stomata control is changed in a chlorophyll b-free barley mutant.

The barley (Hordeum vulgare L.) chlorina f2 3613 mutant exhibits low photosynthesis and slow growth. This results from downregulation of the levels of photosynthetic antenna proteins caused by the absence of chl b, the major regulator of photosynthetic antennae in land plants. Here, we demonstrate that, when grown in the field in full sunlight, this mutant displays a changed pattern of stomatal responses compared with the parental wild-type cultivar Donaria. However, stomatal regulation of chlorina f2 3613 plants was restored when plants were placed under a shade cover for several days. The shade cover reduced incident PAR from 2000-2200μmolm-2s-1 to 800-880μmolm-2s-1 as measured at noon. Contents of ABA, the xanthophyll precursors of ABA biosynthesis and minor antenna proteins, as well as reactive oxygen species levels in stomata and the sensitivity of stomata to exogenously supplied ABA, were determined in leaves of wild-type Donaria and chlorina f2 3613 before and after shading. The results support the view that the restoration of stomatal control in barley chlorina f2 3613 is correlated with an increase in the levels of the minor antenna protein Lhcb6, which has recently been implicated in the enhancement of stomatal sensitivity to ABA in Arabidopsis thaliana (L.) Heynh.

The absence of chlorophyll b affects lateral mobility of photosynthetic complexes and lipids in grana membranes of Arabidopsis and barley chlorina mutants.

The lateral mobility of integral components of thylakoid membranes, such as plastoquinone, xanthophylls, and pigment-protein complexes, is critical for the maintenance of efficient light harvesting, high rates of linear electron transport, and successful repair of damaged photosystem II (PSII). The packaging of the photosynthetic pigment-protein complexes in the membrane depends on their size and stereometric parameters which in turn depend on the composition of the complexes. Chlorophyll b (Chlb) is an important regulator of antenna size and composition. In this study, the lateral mobility (the mobile fraction size) of pigment-protein complexes and lipids in grana membranes was analyzed in chlorina mutants of Arabidopsis and barley lacking Chlb. In the Arabidopsis ch1-3 mutant, diffusion of membrane lipids decreased as compared to wild-type plants, but the diffusion of photosynthetic complexes was not affected. In the barley chlorina f2 3613 mutant, the diffusion of pigment-protein complexes significantly decreased, while the diffusion of lipids increased, as compared to wild-type plants. We propose that the size of the mobile fractions of pigment-protein complexes in grana membranes in vivo is higher than reported previously. The data are discussed in the context of the protein composition of antennae, characteristics of the plastoquinone pool, and production of reactive oxygen species in leaves of chlorina mutants.

Phenolic compounds and carotenoids during acclimation of spring barley and its mutant Chlorina f2 from high to low irradiance

We examined the dynamics of phenolic compounds (PheCs) and carotenoids (Cars) in the leaves of wild type (WT) spring barley (Hordeum vulgare L.) and its mutant lacking chlorophyll b Chlorina f2 (Clo f2) grown from seeds at high irradiance (8 d at 1 000 μmol m-2 s-1; HI) during 9 d of acclimation to low irradiance (50 μmol m-2 s-1; LI). Our results show that a leaf epidermal flavonoid UV-shielding index remained rather constant after transfer of plants from HIto LIconditions and that it was significantly lower in Clo f2 compared to WTplants. This suggests that HIpretreated plants can be well protected against excessive UVfor at least 9 d, as supported also by the constant absorbance of leaf PheCs extracts in the UV-A region (at 335 nm). In contrast, absorbance in the UV-B region (at 270 nm) was reduced, particularly during the initial days of LItreatment, indicating specific changes in PheC profile. High-performance liquid chromatography of soluble PheCs revealed stable content of the major PheC saponarin during LIacclimation, whereas luteolin and feruloylquinic acid content decreased, particularly in WTplants. We also observed a pronounced decrease in Car relative content, particularly a reduction in the xanthophyll cycle pigments (violaxanthin, antheraxanthin, and zeaxanthin, VAZ) pool and diminution of their de-epoxidation state (DEPS) in dark-adapted leaves. As both VAZ and DEPS were higher in HI-acclimated Clo f2 plants than they were in WTplants, the presence of a significant VAZ pool within the lipid phase of thylakoid membrane is indicated. That can contribute to antioxidant capacity particularly in Clo f2 plants. We can therefore conclude that there is a tendency to retain the PheCs responsible for UVshielding during LIacclimation. Meanwhile, the accumulation of both PheCs and zeaxanthin serving as effective antioxidants is considerably downregulated within 9 d.

The chl a fluorescence intensity is remarkably insensitive to changes in the chlorophyll content of the leaf as long as the chl a/b ratio remains unaffected

The effects of changes in the chlorophyll (chl) content on the kinetics of the OJIP fluorescence transient were studied using two different approaches. An extensive chl loss (up to 5-fold decrease) occurs in leaves suffering from either an Mg2+ or SO42− deficiency. The effects of these treatments on the chl a/b ratio, which is related to antenna size, were very limited. This observation was confirmed by the identical light intensity dependencies of the K, J and I-steps of the fluorescence rise for three of the four treatments and by the absence of changes in the F685 nm/F695 nm-ratio of fluorescence emission spectra measured at 77K. Under these conditions, the F0 and FM-values were essentially insensitive to the chl content. A second experimental approach consisted of the treatment of wheat leaves with specifically designed antisense oligodeoxynucleotides that interfered with the translation of mRNA of the genes coding for chl a/b binding proteins. This way, leaves with a wide range of chl a/b ratios were created. Under these conditions, an inverse proportional relationship between the FM values and the chl a/b ratio was observed. A strong effect of the chl a/b ratio on the fluorescence intensity was also observed for barley Chlorina f2 plants that lack chl b. The data suggest that the chl a/b ratio (antenna size) is a more important determinant of the maximum fluorescence intensity than the chl content of the leaf.

Absence of the major light-harvesting antenna proteins alters the redox properties of photosystem II reaction centres in thechlorina F2mutant of barley

Although the chlorina F2 mutant of barley specifically exhibits reduced levels of the major light-harvesting polypeptides associated with photosystem II (PSII), thermoluminescence measurements of photosystem reaction centre photochemistry revealed that S2/S3QB- charge recombinations were shifted to lower temperatures, while the characteristic peak of S2QA- charge recombinations was shifted to higher temperatures compared with wild-type (WT) barley. Thus, we show that the absence of the major light-harvesting polypeptides affects the redox properties of PSII reaction centres. Radiolabeling studies in vivo and in vitro with [32P]orthophosphate or [gamma-32P]ATP, respectively, demonstrated that the D1 PSII reaction centre polypeptide is phosphorylated in both the WT and the F2 mutant. In contrast with the radiolabeling results, phosphorylation of D1 and other PSII proteins, although detected in WT barley, was ambiguous in the F2 mutant when the phosphothreonine antibody method of detection was used. Thus, caution must be exercised in the use of commercially available phosphothreonine antibodies to estimate thylakoid polypeptide phosphorylation. Furthermore, in membrano, the D1 polypeptide of the F2 mutant was less susceptible to trypsin treatment than that of WT barley. The role of the light-harvesting complex in modulating the structure and function of the D1 polypeptide of PSII reaction centers is discussed.

Kinetic Model of Electron-Transport Reactions in Thylakoid Membranes Determining Chlorophyll Fluorescence Transients

ABSTRACTThe analysis of chlorophyll fluorescence transients is very informative approach for evaluation of the physiological state of the photosynthetic machinery of higher plants. Tsimilli-Mishael and Strasser (2008) proposed a system (JIP-test) using the fluorescence transients for calculation of values of energy flows and yields at different stages of the photosynthetic process. In our paper a new model approach is developed for determination of electron transfer rate constants for different sites of the electron transfer chain using the same chlorophyll fluorescence transients. Two kinetic models of the photosynthetic process are constructed. The first one simulates the JIP-test and describes processes determining the three phases of the fluorescence transients during the first second of induction: primary photochemical reaction, electron transfer in the acceptor sides of Photosystem II (PS II) and of PS I. The second model includes most of the components of the electron transfer chain from PS II donor side to PS I. The fitting of the model to real transients allows the determination of rate constant values. Special software Modelyzer for numerical analysis of models is developed. The models are used for analysis of leaves from control plants and others with photosynthetic activity modified as a result of inhibition of electron transport by the herbicide 3-(3,4- dichlorophenil)-1,1-dimethylurea in barley plants—wild type and chlorophyll b-less Chlorina f2 mutant. A good correlation between model values of rate constants with JIP-test results is observed.

The lack of LHCII proteins modulates excitation energy partitioning and PSII charge recombination in Chlorina F2 mutant of barley

Analysis of the partitioning of absorbed light energy within PSII into fractions utilized by PSII photochemistry (ØPSII), thermally dissipated via ΔpH-and zeaxanthin-dependent energy quenching (ØNPQ) and constitutive non-photochemical energy losses (ØNO) was performed in wild type and F2 mutant of barley. The estimated energy partitioning of absorbed light to various pathways indicated that the fraction of ØPSII was slightly higher, while the proportion of thermally dissipated energy through ØNPQ was 38% lower in F2 mutant than in WT. In contrast, ØNO, i.e. the fraction of absorbed light energy dissipated by additional quenching mechanism(s) was 34% higher in F2 mutant. The increased proportion of ØNO correlated with narrowing the temperature gap (ΔT M) between S2/3QB- and S2QA- charge recombinations in F2 mutant as revealed by thermoluminescence measurements. We suggest that this would result in increased probability for an alternative non-radiative P680+QA- radical pair recombination pathway for energy dissipation within the reaction centre of PSII (reaction center quenching) and that this additional quenching mechanism might play an important role in photoprotection when the capacity for the primary, zeaxanthin-dependent non-photochemical quenching (ØNPQ) and state transitions pathways are restricted in the absence of LHCII polypeptides in F2 mutant.

Epidermal UV-shielding and photosystem II adjustment in wild type and chlorina f2 mutant of barley during exposure to increased PAR and UV radiation

The wild-type barley (WT; Hordeum vulgare L.) and its chlorophyll (Chl) b-less mutant chlorina f2 ( clo f2) grown under shaded conditions in a greenhouse were transferred to outdoor conditions in early June with predominantly bright sunny days. During 6 days following transfer of plants we monitored the content of photosynthetic pigments, functional state of photosystem II (PSII) by means of Chl fluorescence induction kinetics and epidermal UV-shielding efficiency using Chl fluorescence imaging technique. Clo f2 mutant was more sensitive to exposure to an enhanced natural solar irradiance than WT barley. Nevertheless, clo f2 as well as WT were able to cope with stressful outdoor conditions, as was documented by the recovery of Chl a content and the maximal photochemical efficiency of PSII ( F V/ F M) after an initial decline. This was due to the immediate carotenoid-mediated photoprotection, reflected by strongly increased total carotenoids content and thermal energy dissipation localized within light-harvesting complexes of PSII (assessed by non-photochemical quenching of minimal fluorescence level). The positive acclimation response was further documented by an enhanced light-saturated electron transport rate through PSII (ETR). Based on the ratios of blue- to UV-excited Chl fluorescence we found that for both WT and clo f2 epidermal UV-shielding increased clearly after transfer to outdoor conditions and reached a saturation level after 3 days. In comparison with WT, clo f2 exhibited lower ability to induce UV-shielding. The kinetics of UV-shielding development during the outdoor treatment was different for the particular leaf regions. We suggest that this is related to the different age and developmental stage of the tissue along the leaf blade. The complementarity of carotenoid-mediated photoprotection and UV-shielding in acclimation of the assimilatory apparatus to increased visible and UV radiation is discussed.

Acclimation to temperature and irradiance modulates PSII charge recombination

Acclimation of wild type and the chlorina F2 mutant of barley to either high light or low temperature results in a 2- to 3-fold increase in non-photochemical quenching which occurred independently of either energy-dependent quenching (qE), xanthophyll cycle-mediated antenna quenching or state transitions. Results of in vivo thermoluminescence measurements used to address this conundrum indicated that excitation pressure regulates the temperature gap for S 2 Q B - and S 2 Q A - charge recombinations within photosystem II reaction centers. This is discussed in terms of photoprotection through non-radiative charge recombination.

Relationship between the organization of the PSII supercomplex and the functions of the photosynthetic apparatus

The chlorophyll fluorescence and the photosynthetic oxygen evolution (flash-induced oxygen yield patterns and oxygen bursts under continuous irradiation) were investigated in the thylakoid membranes with different stoichiometry and organization of the chlorophyll-protein complexes. Data show that the alteration in the organization of the photosystem II (PSII) supercomplex, i.e. the amount and the organization of the light-harvesting chlorophyll a/ b protein complex (LHCII), which strongly modifies the electric properties of the membranes, influences both the energy redistribution between the two photosystems and the oxygen production reaction. The decrease of surface electric parameters (charge density and dipole moments), associated with increased degree of LHCII oligomerization, correlates with the strong reduction of the energy transfer from PSII to PSI. In the studied pea thylakoid membranes (wild types Borec, Auralia and their mutants Coeruleovireus 2/ 16, Costata2/ 133, Chlorotica XV/1422) with enhanced degree of oligomerization of LHCII was observed: (i) an increase of the S 0 populations of PSII in darkness; (ii) an increase of the misses; (iii) an alteration of the decay kinetics of the oxygen bursts under continuous irradiation. There is a strict correlation between the degree of LHCII oligomerization in the investigated pea mutants and the ratio of functionally active PSIIα to PSIIβ centers, while in thylakoid membranes without oligomeric structure of LHCII ( Chlorina f2 barley mutant) the PSIIα centers are not registered.

Effects of different excitation and detection spectral regions on room temperature chlorophyll a fluorescence parameters

The effects of different spectral region of excitation and detection of chlorophyll (Chl) a fluorescence at room temperature on the estimation of excitation energy utilization within photosystem (PS) 2 were studied in wild-type barley (Hordeum vulgare L. cv. Bonus) and its Chl b-less mutant chlorina f2 grown under low and high irradiances [100 and 1 000 µmol(photon) m−2 s−1]. Three measuring spectral regimes were applied using a PAM 101 fluorometer: (1) excitation in the red region (maximum at the wavelength of 649 nm) and detection in the far-red region beyond 710 nm, (2) excitation in the blue region (maximum at the wavelength of 461 nm) and detection beyond 710 nm, and (3) excitation in the blue region and detection in the red region (660– 710 nm). Non-photochemical quenching of maximal (NPQ) and minimal fluorescence (SV0), determined by detecting Chl a fluorescence beyond 710 nm, were significantly higher for blue excitation as compared to red excitation. We suggest that this results from higher non-radiative dissipation of absorbed excitation energy within light-harvesting complexes of PS2 (LHC2) due to preferential excitation of LHC2 by blue radiation and from the lower contribution of PS1 emission to the detected fluorescence in the case of blue excitation. Detection of Chl a fluorescence originating preferentially from PS2 (i.e. in the range of 660–710 nm) led to pronounced increase of NPQ, SV0, and the PS2 photochemical efficiencies (FV/FM and FV′/FM′), indicating considerable underestimation of these parameters using the standard set-up of PAM 101. Hence PS1 contribution to the minimal fluorescence level in the irradiance-adapted state may reach up to about 80 %.

The effect of outer antenna complexes on the photochemical trapping rate in barley thylakoid Photosystem II

We have investigated the previous suggestions in the literature that the outer antenna of Photosystem II of barley does not influence the effective photosystem primary photochemical trapping rate. It is shown by steady state fluorescence measurements at the F 0 fluorescence level of wild type and the chlorina f2 mutant, using the chlorophyll b fluorescence as a marker, that the outer antenna is thermally equilibrated with the core pigments, at room temperature, under conditions of photochemical trapping. This is in contrast with the conclusions of the earlier studies in which it was suggested that energy was transferred rapidly and irreversibly from the outer antenna to the Photosystem II core. Furthermore, the effective trapping time, determined by single photon counting, time-resolved measurements, was shown to increase from 0.17±0.017 ns in the chlorina Photosystem II core to a value within the range 0.42±0.036–0.47±0.044 ns for the wild-type Photosystem II with the outer antenna system. This 2.5–2.8-fold increase in the effective trapping time is, however, significantly less than that expected for a thermalised system. The data can be explained in terms of the outer antenna increasing the primary charge separation rate by about 50%.

Regulation of the excitation energy utilization in the photosynthetic apparatus of chlorina f2 barley mutant grown under different irradiances

Acclimation of the photosynthetic apparatus of chlorophyll b-less barley mutant chlorina f2 to low light (100 μmol m −2 s −1; LL) and extremely high light level (1000 μmol m −2 s −1; HL) was examined using techniques of pigment analysis and chlorophyll a fluorescence measurements at room temperature and at 77 K. The absence of chlorophyll b in LL-grown chlorina f2 resulted in the reduction of functional antenna size of both photosystem II (by 67%) and photosystem I (by 21%). Chlorophyll fluorescence characteristics of the LL-grown mutant indicated no impairment of the utilization of absorbed light energy in photosystem II photochemistry. Thermal dissipation of excitation energy estimated as non-photochemical quenching of minimal fluorescence (SV 0) was significantly higher as compared to the wild-type barley grown under LL. Despite impaired assembly of pigment–protein complexes, chlorina f2 was able to efficiently acclimate to HL. In comparison with chlorina f2 grown under LL, HL-grown chlorina f2 was characterized by unaffected maximal photochemical efficiency of photosystem II ( F V/ F M), doubled content of both β-carotene and the xanthophyll cycle pigments and considerably reduced efficiency of excitation energy transfer from carotenoids to chlorophyll a. The enormous xanthophyll cycle pool size was however associated with reduced SV 0 capacity. We suggest that the substantial part of the xanthophyll cycle pigments is not bound to the remaining pigment–protein complexes and acts as filter for excitation energy, thereby contributing to the efficient photoprotection of chlorina f2 grown under HL.

Simultaneous analysis of prompt and delayed chlorophyll a fluorescence in leaves during the induction period of dark to light adaptation

An attempt is made to reveal the relation between the induction curves of delayed fluorescence (DF) registered at 0.35–5.5 ms and the prompt chlorophyll fluorescence (PF). A simple formulation was proposed to link the ratio of the transient values of delayed and variable fluorescence with the redox state of the primary electron acceptor of Photosystem II—Q A, and the thylakoid membrane energization. The term luminescence potential ( U L ) was introduced, defined as the sum of the redox potential of Q A and the transmembrane proton gradient. It was shown that U L is proportional to the ratio of DF to the variable part of PF. The theoretical model was verified and demonstrated by analysing induction courses of PF and millisecond DF, simultaneously registered from leaves of barley—wild-type and the chlorophyll b-less mutant chlorina f2. A definitive correlation between PF and DF was established. If the luminescence changes are strictly due to U L , the courses of DF and PF are reciprocal and the millisecond DF curve resembles the first derivative of the PF( t) function.

Response of chlorina barley mutants to heat stress under low and high light.

Barley plants (Hordeum vulgare L.) of wild type and two chlorina mutants, chlorina 126 and chlorina f2, were subjected to 42°C for 5 h at light intensities of 100 and 1000 μmol photons m-2 s-1. The exposure of plants to heat stress at a light intensity of 100 μmol m-2 s-1 induced enormous proline accumulation, indicating that the effect of heat stress was stronger when it was combined with low light intensity. The functional activity of PSII, O2evolution and flash-induced thermoluminescence B-band amplitude were strongly reduced when plants were exposed to heat at low light intensity. The results clearly showed that high light intensity had a protective effect on photosynthetic activity when barley plants were treated with high temperature. Comparison of the thermosensitivity of wild type plants and chlorina mutants revealed that O2 evolution in chlorina 126 and, especially, in chlorina f2 was more sensitive to heat than in wild type.

Spectral characterization of chlorophyll fluorescence in barley leaves during linear heating: Analysis of high-temperature fluorescence rise around 60°C

The spectral characteristics of chlorophyll fluorescence and absorption during linear heating of barley leaves within the range 25–75°C (fluorescence temperature curve, FTC) were studied. Leaves with various content of light harvesting complexes (green, Chl b-less chlorina f2 and intermittent light grown) revealing different types of FTC were used. Differential absorption, emission and excitation spectra documented four characteristic phases of the FTC. The initial two FTC phases (a rise in the 46–49°C region and a subsequent decrease to about 55°C) mostly reflected changes in the fluorescence quantum yield peaking at about 685 nm. A steep second fluorescence rise at 55–61°C was found to originate from a short-wavelength Chl a spectral form (emission maximum at 675 nm) causing a gradual blue shift of the emission spectra. In this temperature range, a clear correspondence of the blue shift in the emission and absorption spectra was found. We suggest that the second fluorescence rise in FTC reflects a weakening of the Chl a–protein interaction in the thylakoid membrane.

Global spectral-kinetic analysis of room temperature chlorophyll a fluorescence from light-harvesting antenna mutants of barley.

This study presents a novel measurement, and simulation, of the time-resolved room temperature chlorophyll a fluorescence emission spectra from leaves of the barley wild-type and chlorophyll-b-deficient chlorina (clo) f2 and f104 mutants. The primary data were collected with a streak-camera-based picosecond-pulsed fluorometer that simultaneously records the spectral distribution and time dependence of the fluorescence decay. A new global spectral-kinetic analysis programme method, termed the double convolution integral (DCI) method, was developed to convolve the exciting laser pulse shape with a multimodal-distributed decay profile function that is again convolved with the spectral emission band amplitude functions. We report several key results obtained by the simultaneous spectral-kinetic acquisition and DCI methods. First, under conditions of dark-level fluorescence, when photosystem II (PS II) photochemistry is at a maximum at room temperature, both the clo f2 and clo f104 mutants exhibit very similar PS II spectral-decay contours as the wild-type (wt), with the main band centred around 685 nm. Second, dark-level fluorescence is strongly influenced beyond 700 nm by broad emission bands from PS I, and its associated antennae proteins, which exhibit much more rapid decay kinetics and strong integrated amplitudes. In particular a 705-720 nm band is present in all three samples, with a 710 nm band predominating in the clo f2 leaves. When the PS II photochemistry becomes inhibited, maximizing the fluorescence yield, both the clo f104 mutant and the wt exhibit lifetime increases for their major distribution modes from the minimal 205-500 ps range to the maximal 1500-2500 ps range for both the 685 nm and 740 nm bands. The clo f2 mutant, however, exhibits several unique spectral-kinetic properties, attributed to its unique PS I antennae and thylakoid structure, indicating changes in both PS II fluorescence reabsorption and PS II to PS I energy transfer pathways compared to the wt and clo f104. Photoprotective energy dissipation mediated by the xanthophyll cycle pigments and the PsbS protein was uninhibited in the clo f104 mutant but, as commonly reported in the literature, significantly inhibited in the clo f2; the inhibited energy dissipation is partly attributed to its thylakoid structure and PS II to PS I energy transfer properties. It is concluded that it is imperative with steady-state fluorometers, especially for in vivo studies of PS II efficiency or photoprotective energy dissipation, to quantify the influence of the PS I spectral emission.

Contribution of LHC II complex to the electric properties of thylakoid membranes: an electric light scattering study of Chl b-less barley mutant

Electric light scattering measurements demonstrate a strong decline in the permanent electric dipole moment and electric polarizability of both thylakoid membranes and photosystem II-enriched particles of the Chlorina f2 mutant which has severely reduced levels of light-harvesting chlorophyll a/ b-binding proteins compared to the wild type barley chloroplasts. The shift in the electric polarizability relaxation to higher frequencies in thylakoids and photosystem II particles from Chlorina f2 reflects higher mobility of the interfacial charges of the mutant than that of the wild type membranes. The experimental data strongly suggest that the major light-harvesting complex of photosystem II directly contribute to the electric properties of thylakoid membranes.

Greening under high light or cold temperature affects the level of xanthophyll-cycle pigments, early light-inducible proteins, and light-harvesting polypeptides in wild-type barley and the chlorina f2 mutant

Etiolated seedlings of wild type and the chlorina f2 mutant of barley (Hordeum vulgare) were exposed to greening at either 5 degrees C or 20 degrees C and continuous illumination varying from 50 to 800 &mgr;mol m-2 s-1. Exposure to either moderate temperature and high light or low temperature and moderate light inhibited chlorophyll a and b accumulation in the wild type and in the f2 mutant. Continuous illumination under these greening conditions resulted in transient accumulations of zeaxanthin, concomitant transient decreases in violaxanthin, and fluctuations in the epoxidation state of the xanthophyll pool. Photoinhibition-induced xanthophyll-cycle activity was detectable after only 3 h of greening at 20 degrees C and 250 &mgr;mol m-2 s-1. Immunoblot analyses of the accumulation of the 14-kD early light-inducible protein but not the major (Lhcb2) or minor (Lhcb5) light-harvesting polypeptides demonstrated transient kinetics similar to those observed for zeaxanthin accumulation during greening at either 5 degrees C or 20 degrees C for both the wild type and the f2 mutant. Furthermore, greening of the f2 mutant at either 5 degrees C or 20 degrees C indicated that Lhcb2 is not essential for the regulation of the xanthophyll cycle in barley. These results are consistent with the thesis that early light-inducible proteins may bind zeaxanthin as well as other xanthophylls and dissipate excess light energy to protect the developing photosynthetic apparatus from excess excitation. We discuss the role of energy balance and photosystem II excitation pressure in the regulation of the xanthophyll cycle during chloroplast biogenesis in wild-type barley and the f2 mutant.

Effects of ultraviolet-B light on photosystem II phosphoproteins in barley wild type and its chlorophyll b-less mutant chlorina f2

The effects of ultraviolet-B light on the level and steady-state phosphorylation of photosystem II proteins have been studied in barley wild type and its chlorophyll b-less mutant chlorina f2. In the wild type, ultraviolet-B radiation is found to promote dephosphorylation of all thylakoid phosphoproteins. In addition, for reaction-centre proteins D1 and D2, dephosphorylation is paralleled by degradation. Photosystem II core proteins in the mutant are not found to be significantly phosphorylated in any experimental conditions, and loss of D1 and D2 reaction-centre proteins is slightly faster than in the wild type. These results are consistent with the possibility that phosphorylation of reaction-centre proteins affects their stability, possibly by slowing down the rate of degradation, as in the case of visible light.