Contrasting modes of regulation of PS II light utilization with changing irradiance in normal and psbS mutant leaves of Arabidopsis thaliana

Complementary techniques of chlorophyll a fluorescence, steady state CO₂ exchange, and O₂ release during a multiple turnover flash were applied to compare responses to irradiance for leaves of wild type and psbS mutants. The latter included variants in which the psbS gene was deleted (npq4-1) or possessed a single point mutation (npq4-9). Nonphotochemical quenching (NPQ) was reduced by up to 80 and 50%, respectively, in these lines at high irradiance. Analysis of changes in steady-state fluorescence yields and quantum yield of linear electron transport in the context of the reversible radical pair model of Photosystem II (PS II) indicated that NPQ occurs by nonradiative deactivation of chlorophyll singlet states in normal leaves. In contrast, application of the same criteria together with the observed irreversibility of NPQ and decline in density of functional PS II reaction centers following excessive illumination indicated a change in reaction center properties for the psbS deletion phenotype (Npq4-1^–). Specifically, PS II reaction centers in Npq4-1^– convert to a photochemically inactive, yet strongly quenching, form in intense light. The possibility of formation of a carotenoid or chlorophyll cation quencher in the reaction center is discussed. Results for the point mutant phenotype (Npq4-9^–) were intermediate to those of wild-type and Npq4-1^–. Furthermore, wild-type leaves exhibited a significant reversible increase in the PS II in vivo rate constant for photochemistry (k_P0) in saturating compared to limiting light. Changes in k_P0 could not be accounted for in terms of a classic phosphorylation-dependent (state transition) mechanism. Changes in k_P0 may arise from alternate pigment—protein conformations that alter the way excitons equilibrate among PS II chromophores. The lack of similar irradiance-dependent changes in k_P0 for the psbS mutants suggests a role for the PS II-S protein in the regulation of exciton distribution.This revised version was published online in October 2005 with corrections to the Cover Date.     

Measurement of chlorophyll fluorescence within leaves using a modified PAM Fluorometer with a fiber-optic microprobe

By using a fiber-optic microprobe in combination with a modified PAM Fluorometer, chlorophyll fluorescence yield was measured within leaves with spatial resolution of approximately 20 m. The new system employs a miniature photomultiplier for detection of the pulse-modulated fluorescence signal received by the 20 m fiber tip. The obtained signal/noise ratio qualifies for recordings of fluorescence induction kinetics (Kautsky effect), fluorescence quenching by the saturation pulse method and determination of quantum yield of energy conversion at Photosystem II at different sites within a leaf. Examples of the system performance and of practical applications are given. It is demonstrated that the fluorescence rise kinetics are distinctly faster when chloroplasts within the spongy mesophyll are illuminated as compared to palisade chloroplasts. Photoinhibition is shown to affect primarily the quantum yield of the palisade chloroplasts when excessive illumination is applied from the adaxial leaf side. The new system is envisaged to be used in combination with light measurements within leaves for an assessment of the specific contributions of different leaf regions to overall photosynthetic activity and for an integrative modelling of leaf photosynthesis.This paper is dedicated to Ulrich Heber on the occasion of his 65th birthday, with great respect for his outstanding achievements in photosynthesis research.     

Photosynthesis and photoinhibition in leaves of chlorophyll b-less barley in relation to absorbed light

The response of photosynthesis to absorbed light by intact leaves of wild-type ( Hordeum vulgare L. cv. Gunilla) and chlorophyll b -less barley ( H. vulgare L. cv. Dornaria, chlorina-f2²⁸⁰⁰) was measured in a light integrating sphere. Up to the section where the light response curve bends most sharply the responses of the b -less and wild-type barley were similar but not identical. Average quantum yield and convexity for the mutant light response curves were 0.89 and 0.90, respectively, times those of the wild-type barley. The maximum quantum yield for PSII photochemistry was also 10% lower as indicated by fluorescence induction kinetics (F_v/F_m). Just above the region where the light curve bends most sharply, photosynthesis decreased with time in the mutant but not in the wild-type barley. This decrease was associated with a decrease in F_v/F_m indicating photoinhibition of PSII. This photoinhibition occurred in the same region of the light response curve where zeaxanthin formation occurs. Zeaxanthin formation occurred in both the chlorophyll b -less and wild-type leaves. However, the epoxidation state was lower in the mutant than in the wild-type barley. The results indicate that chlorophyll b -less mutants will have reduced photosynthetic production as a result of an increased sensitivity to photoinhibition and possibly a lowered quantum yield and convexity in the absence of photoinhibition.     

Tropical lotic primary producers: Who has the most efficient photosynthesis in low‐order stream ecosystems?

1. Photosynthetic performance was assessed for the main groups of primary producers of low‐order tropical streams, comprising macroalgae (11 taxa of Chlorophyta, Rhodophyta and Xanthophyta plus Cyanobacteria), bryophytes (2), angiosperms (3), and the periphytic community, by means of the Chl‐a fluorescence and oxygen evolution techniques.
2. Samples of primary producer were collected in nine stream reaches within the same drainage basin that were characterised as open or partially shaded.
3. Low values of compensation irradiance (I_c) were found for all species tested, suggesting that they are able to maintain an autotrophic metabolism even under very low irradiances.
4. Parameters derived from photosynthesis–irradiance curves revealed high values of the saturation parameter (I_k), associated with relative low values of photosynthetic efficiency (αETR) and photoinhibition (βETR) for species of Chlorophyta and the periphyton, suggesting adaptation to high irradiance conditions.
5. Species of Rhodophyta, Xanthophyta, Bryophyta and angiosperms showed, in general low values of I_k and moderate to high values of αETR and βETR, indicating typical photosynthetic responses of shade‐adapted plants.
6. Chlorophyta and the periphyton showed high photosynthetic efficiency in high irradiance, and rhodophytes, bryophytes and angiosperms seemed to be more efficient in shaded stream stretches.
7. Considering that Cyanobacteria showed high values of the photosynthesis/respiration (P/R) ratio, we suggest that this group is the most efficient primary producer in a wide range of low‐order tropical streams. This physiological trait could be a feature that favours the abundance of Cyanobacteria in the benthic primary producer community, as is often reported in the literature.

     

GROWTH AND PHOTOSYNTHESIS OF ULVA ROTUNDATA (CHLOROPHYTA) AS A FUNCTION OF TEMPERATURE AND SQUARE WAVE IRRADIANCE IN INDOOR CULTURE1

Temperature and photon flux density (PFD) vary independently in estuaries, e.g. high PFD may occur at any temperature, so it is necessary to consider synergistic effects of these factors on algal growth. Because natural PFD is highly variable and daylength changes confound seasonal temperature cycles, it is easier to interpret factorial experiments in controlled laboratory conditions. Clonal Ulva rotundata Blid. (Chlorophyta) has been studied extensively in outdoor culture. In this study it was maintained indoors under square wave photoperiods at five PFDs and three temperatures. Growth rate, photqsynthetic light response (P-I) curves, and photosystem II chlorophyll fluorescence properties were measured at the growth temperature following acclimation. Interactions between PFD and growth temperature were strongly indicated in all physiological parameters measured. Greatest PFD response occurred at the highest temperature, and the largest temperature response occurred at the highest PFD. Light-saturated photosynthesis (P_m) dark respiration (R_d), and light-limited quantum yield (Φ_m) were sufficient to describe acclimation status. The light-saturation parameter (I_k) was redundant and potentially misleading. Although U. rotundata exhibits a great amplitude of photoacclimation, it apparently has little capacity for temperature acclimation compared to the kelp, Laminaria saccharina, for which published data indicate similar photosynthetic rates over a broad range of growth temperatures. Diurnal variation of P_m and R_d at a growth PFD of ～ 1700 ± 200 μmol photons · m^?2· s^?1 was similar to the pattern observed previously in outdoor culture, suggesting endogenous control of these parameters. Quantum yield and the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m), which were depressed in midday sunlight exceeding ～ 1500 μmol photons · m^?2· s^?1, were relatively invariant through the day in indoor culture, indicating that these parameters are controlled primarily by instantaneous PFD. Growth and fluorescence data are also presented for some other macroalgae for comparative purposes.     

油菜叶片气体交换对O３浓度和熏蒸方式的响应

运用CIRAS-1型便携式光合作用测定系统,在田间原位比较研究了不同O3浓度(CF,50 nl.L-1和100 nl.L-1)和熏蒸方式(恒定和动态)油菜叶片的气体交换特征及其对光强、CO2浓度升高的响应。结果表明(1)恒定熏气下,O3浓度增加导致叶片的蒸腾速率降低,水分利用效率提高,但动态熏蒸则引起蒸腾速率增加,水分利用效率下降,而且明显导致光合速率和气孔导度的降低;(2)高浓度的O3(100 nl.L-1)引起叶片的表观量子产额、暗呼吸饱和光强和最大净光合速率显著降低,光呼吸和CO2补偿点显著升高;熏蒸方式对叶片的暗呼吸、光补偿点、饱和光强、最大光合速率、羧化效率的影响差异显著;(3)不论何种熏蒸方式,高浓度的O3都引起下叶位的Fv/Fo、Fv/Fm显著降低,对上叶位没有影响。相同剂量下,动态熏蒸对叶片气体交换的影响更大,不利于植物生长和干物质的积累。     

PHOTOSYNTHETIC ACTIVITY OF TWO DEVELOPMENTAL STAGES OF A NOSTOC STRAIN (CYANOBACTERIA) ISOLATED FROM GEOSIPHON PYRIFORME (MYCOTA)1

Sessile colonies and motile hormogonia, the two main developmental stages in the life cycle of a Nostoc strain isolated from the endocytobiosis with Geosiphon pyriforme (Kützing) F. v. Wettstein, were investigated for their photosynthetic competence. Large-scale fractionation of the two stages is presented. Photosynthetic parameters were assessed by measurement of chlorophyll fluorescence and oxygen evolution. Hormogonia were as photosynthetically competent as the colonial stage. In addition, hormogonia showed an enhanced capability for nonradiative dissipation of absorbed light energy, a feature that might be important for their function as propagula. Data for the quantum yield of photosystem II of the isolated Nostoc strain were compared to the values determined in situ in G. pyriforme and indicated the possibility of a higher photosynthetic capacity of the endosymbiotic as compared to the isolated cyanobacterium.     

The fluorescence lifetime and quantum yield of chlorophyll a in Triton X-100 solutions

The fluorescence of chlorophyll (Chl) a in 0.007–0.1% Triton X-100 was investigated by a phase-shift technique. The Chl a concentrations varied from 0.7 to 25 μM. Parallel measurements of fluorescence lifetime (τ) and quantum yield (ψ) were made. It was concluded that homogeneous energy transfer takes place at detergent concentrations above 0.025%: (i) the transfer between uniform molecules of the pigment solubilized in Triton X-100 micelles, when τ and ψ are constant; (ii) the transfer towards the quenching centers, resulting in a proportional decrease in τ and ψ. At a Triton X-100 concentration of about 0.025% the Chl a emission becomes heterogeneous. It is evident from the disproportional decrease in τ and ψ (greater in ψ than in τ) and also from the rise of the fluorescence at 730–750 nm. As the Triton X-100 concentration becomes lower than the critical one (0.021%), the number of micelles drops abruptly and Chl a forms colloid particles in the aqueous medium. This manifests itself as a decrease in τ and as a certain stabilization of ψ. Having analyzed the complex pattern of the τ/ψ ratio, we concluded that under these conditions more than 90% of Chl a is in a weakly fluorescent form (τ < 30 ps) and about 1% is in an aggregated state fluorescing at 732 nm with τ about 0.7 ns.     

Evaluation of instant light‐response curves of chlorophyll fluorescence parameters obtained with a portable chlorophyll fluorometer on site in the field

Miniaturized pulse‐amplitude modulated photosynthesis yield analysers are primarily designed for measuring effective quantum yield (ΔF/F_m′) of photosystem II under momentary ambient light conditions in the field. Although this provides important ecophysiological information, it is often necessary to learn more about the potential intrinsic capacities of leaves by measuring light‐response curves. Thus, instruments provide light‐curve programmes, where light intensities are increased in short intervals and instant light‐response curves are recorded within a few minutes. This method can be criticized because photosynthesis will most likely not be in steady state. This technical report shows that with the appropriate precautions instant light curves can nevertheless provide reliable information about cardinal points of photosynthesis. First, the geometry of the light source of the instrument in relation to the quantum sensor must be considered and quantum sensor readings must be corrected. Second, the measurements of the light‐response curves must be compared with readings of effective quantum yield of photosystem II under ambient light conditions where photosynthesis is in steady state. This may show that in the critical range of the light curves either both measurements perfectly coincide or are offset against each other by a constant value (examples are given here). In the first case results of light curves can be taken at face values, and in the second case a simple correction can be applied. With these precautions and careful interpretations instant light‐response curves can be an enormous advantage in ecophysiological field work.