Kevin Oxborough

Resolving chlorophyll a fluorescence images of photosynthetic efficiency into photochemical and non-photochemical components – calculation of qP and Fv-/Fm-; without measuring Fo-;

Imaging of chlorophyll a fluorescence from leaves has enabled the spatial resolution of the fluorescence parameter, ΔF/Fm-;. Although this parameter provides a reliable estimate of photosynthetic efficiency under most conditions, the extent to which this efficiency is defined by (i) competition with other energy-dissipating processes operating at photosystem II and (ii) by processes on the reducing side of photosystem II, such as carbon assimilation, requires the use of additional parameters. Of particular value are qP, which quantifies the photochemical capacity of photosystem II, and Fv-;/Fm-;, which quantifies the extent to which photochemistry at photosystem II is limited by competition with thermal decay processes. Imaging of both qP and Fv-;/Fm-; requires measurement of Fo-; (the minimum fluorescence yield in the light-adapted state), which cannot be imaged with existing systems. In this paper, a method is described which estimates Fo-; through a simple equation involving the minimum fluorescence yield in the dark-adapted state (Fo), the maximum fluorescence yield in the dark-adapted state (Fm), and the maximum fluorescence yield in the light-adapted state (Fm-;). This method is tested here, through comparison of measured and calculated values of Fo-;. An example of the application of this method to analysis of photosynthetic performance in leaves, from images of chlorophyll a fluorescence, is also presented.

Rapid, Noninvasive Screening for Perturbations of Metabolism and Plant Growth Using Chlorophyll Fluorescence Imaging

A rapid, noninvasive technique involving imaging of chlorophyll fluorescence parameters for detecting perturbations of leaf metabolism and growth in seedlings is described. Arabidopsis seedlings were grown in 96-well microtitre plates for 4 d and then treated with eight herbicides with differing modes of action to induce perturbations in a range of different metabolic processes. Imaging of chlorophyll fluorescence emissions from 96 seedlings growing on a microtitre plate enabled images of a number of fluorescence parameters to be rapidly and simultaneously produced for the plants in each well. Herbicideinduced perturbations in metabolism, even in metabolic reactions not directly associated with photosynthetic metabolism, were detected from the changes in the images of fluorescence parameters considerably before any visual effects on seedling growth were observed. Evaluations of seedling growth were made from measurements of the area of chlorophyll fluorescence emission in images of plants growing in the 96-well plates. Decreased seedling growth related directly to herbicideinduced changes in the imaged chlorophyll fluorescence parameters. The applicability of this rapid-screening technique for metabolic perturbations in monocotyledonous species was demonstrated by treating Agrostis tenuis seedlings with Imazapyr, an inhibitor of branched-chain amino acid synthesis.

Fast repetition rate and pulse amplitude modulation chlorophyll a fluorescence measurements for assessment of photosynthetic electron transport in marine phytoplankton

Pulse amplitude modulation (PAM) and fast repetition rate (FRR) fluorescence are currently used to estimate photosynthetic quantum yields and photosynthetic rates in aquatic systems. Here we compare simultaneous measurements of the photochemical efficiency of photosystem II obtained from the two techniques and independent estimates of the rate of light absorption by photosystem II. We measured the light-dependencies of the photochemical efficiency of photosystem II (Fq′/Fm′) in five phytoplankters using FRR and Xe-PAM approaches. The FRR and PAM estimates were related in a non-linear fashion. At low irradiances, Fq′/Fm′ measured using PAM fluorescence exceeded Fq′/Fm′ measured using FRR fluorescence by about 20%. At high irradiances, measurements of Fq′/Fm′ from the two approaches converged. The differences in Fq′/Fm′ reflect the distinct techniques by which FRR and PAM protocols excite PSII and are amplified when estimating electron transfer rates as a result of the irradiance term. We also found that measurements of the effective light absorption cross-section for photosystem II obtained by FRR fluorescence compared well with estimates obtained from measured light absorption and photosynthetic unit size. Finally, we compared the photon efficiency of gross oxygen evolution from measurements of gross oxygen evolution and light absorption (ΦPO2 18) with FRR measurements of Fq′/Fm′. We found that measurements of Fq′/Fm′ were highly linearly correlated, but were lower by a factor of ∼1.5, than ΦPO2 18.

Factors Associated with Depression of Photosynthetic Quantum Efficiency in Maize at Low Growth Temperature

The photosynthetic productivity of maize (Zea mays) in temperate regions is often limited by low temperatures. The factors responsible for the sensitivity of photosynthesis in maize to growth at suboptimal temperature were investigated by measuring (a) the quantum yields of CO2 fixation and photosystem II (PSII) photochemistry, (b) the pigments of the xanthophyll cycle, (c) the concentrations of active and inactive PSII reaction centers, and (d) the synthesis of core components of PSII reaction centers. Measurements were made on fully expanded leaves grown at 14[deg]C, both before and during the first 48 h after transfer of these plants to 25[deg]C. Our findings indicate that zeaxanthin-related quenching of absorbed excitation energy at PSII is, quantitatively, the most important factor determining the depressed photosynthetic efficiency in 14[deg]C-grown plants. Despite the photoprotection afforded by zeaxanthin-related quenching of absorbed excitation energy, a significant and more persistent depression of photosynthetic efficiency appears to result from low temperature-induced inhibition of the rate at which damaged PSII centers can be replaced.

The trade-off between the light-harvesting and photoprotective functions of fucoxanthin-chlorophyll proteins dominates light acclimation in Emiliania huxleyi (clone CCMP 1516).

Mechanistic understanding of the costs and benefits of photoacclimation requires knowledge of how photophysiology is affected by changes in the molecular structure of the chloroplast. We tested the hypothesis that changes in the light dependencies of photosynthesis, nonphotochemical quenching and PSII photoinactivation arises from changes in the abundances of chloroplast proteins in Emiliania huxleyi strain CCMP 1516 grown at 30 (Low Light; LL) and 1000 (High Light; HL) μmol photons m(-2) s(-1) photon flux densities. Carbon-specific light-saturated gross photosynthesis rates were not significantly different between cells acclimated to LL and HL. Acclimation to LL benefited cells by increasing biomass-specific light absorption and gross photosynthesis rates under low light, whereas acclimation to HL benefited cells by reducing the rate of photoinactivation of PSII under high light. Differences in the relative abundances of proteins assigned to light-harvesting (Lhcf), photoprotection (LI818-like), and the photosystem II (PSII) core complex accompanied differences in photophysiology: specifically, Lhcf:PSII was greater under LL, whereas LI818:PSII was greater in HL. Thus, photoacclimation in E. huxleyi involved a trade-off amongst the characteristics of light absorption and photoprotection, which could be attributed to changes in the abundance and composition of proteins in the light-harvesting antenna of PSII.

The role of herbicides in the erosion of salt marshes in eastern England

Laboratory studies and field trials were conducted to investigate the role of herbicides on saltmarsh vegetation, and their possible significance to saltmarsh erosion. Herbicide concentrations within the ranges present in the aquatic environment were found to reduce the photosynthetic efficiency and growth of both epipelic diatoms and higher saltmarsh plants in the laboratory and in situ. The addition of sublethal concentrations of herbicides resulted in decreased growth rates and photosynthetic efficiency of diatoms and photosynthetic efficiency of higher plants. Sediment stability also decreased due to a reduction in diatom EPS production. There was qualitative evidence that diatoms migrated deeper into the sediment when the surface was exposed to simazine, reducing surface sediment stability by the absence of a cohesive biofilm. Sediment loads on leaves severely reduced photosynthesis in Limonium vulgare. This, coupled with reduced carbon assimilation from the effects of herbicides, could have large negative consequences for plant productivity and over winter survival of saltmarsh plants. The data support the hypothesis that sublethal herbicide concentrations could be playing a role in the increased erosion of salt marshes that has occurred over the past 40 years.

An Integrated Response of Trichodesmium erythraeum IMS101 Growth and Photo-Physiology to Iron, CO2, and Light Intensity

We have assessed how varying CO2 (180, 380, and 720 μatm) and growth light intensity (40 and 400 μmol photons m−2 s−1) affected Trichodesmium erythraeum IMS101 growth and photophysiology over free iron (Fe′) concentrations between 20 and 9,600 pM. We found significant iron dependencies of growth rate and the initial slope and maximal relative PSII electron transport rates (rPm). Under iron-limiting concentrations, high-light increased growth rates and rPm; possibly indicating a lower allocation of resources to iron-containing photosynthetic proteins. Higher CO2 increased growth rates across all iron concentrations, enabled growth to occur at lower Fe′ concentrations, increased rPm and lowered the iron half saturation constants for growth (Km). We attribute these CO2 responses to the operation of the CCM and the ATP spent/saved for CO2 uptake and transport at low and high CO2, respectively. It seems reasonable to conclude that T. erythraeum IMS101 can exhibit a high degree of phenotypic plasticity in response to CO2, light intensity and iron-limitation. These results are important given predictions of increased dissolved CO2 and water column stratification (i.e., higher light exposures) over the coming decades.