The chlorophyll fluorescence (F) temperature curves in a linear time-temperature heating/cooling regime were used to study heat-induced irreversible F changes in primary green leaves of spring barley (Hordeum vulgare L. cv. Akcent). The leaf segments were heated in a stirred water bath at heating rates of 0.0083, 0.0166, 0.0333, and 0.0500 °C s −1 from room temperature up to maximal temperature T m and then linearly cooled to 35 °C at the same rate. The F intensity was measured by a pulse-modulated technique. The results support the existence of the two critical temperatures of irreversible F changes postulated earlier, at 45−48 and 53−55 °C. The critical temperatures are slightly dependent on the heating rate. Two types of parameters were used to characterize the irreversibility of the F changes: the coefficient of irreversibility μ defined as the ratio of F intensity at 35 °C at the starting/ending parts of the cycle and the slopes of tangents of linear parts of the F temperature curve. The dependence of μ on T m revealed a maximum, which moved from 54 to 61 °C with the increasing heating/cooling rate ν from 0.0083 to 0.0500 ºC s −1 , showing two basic phases of the irreversible changes. The Arrhenius and Eyring approaches were applied to calculate the activation energies of the initial increase in μ. The values varied between 30 and 50 kJ mol −1 and decreased slightly with the increasing heating rate.
Measurement of the chlorophyll (Chl) a fluorescence rise (FR) under higher exciting irradiance (EI), the O-J-I-P transient, or under lower irradiance, the O-I-P transient, is a routinely used method to access photosystem 2 function in thylakoid membranes of chloroplasts. Our measurements with a suspension of pea thylakoid membranes showed that the relative heights of the J and I steps in the FR depended not only on EI but also on the concentration and thickness of the sample. We explain this effect as a consequence of the gradient of EI within the sample. We tested this suggestion by theoretical simulations of the FR based on the model that was previously used for simulation of the FR considering in addition the gradient of EI within the sample. Our theoretical results correspond well with the experiments. The irradiance gradient effect may influence measured FR significantly and this fact should be taken into consideration in the interpretation of measured FRs.
In this work, we used barley leaves suffering from a stress, for measurements of chlorophyll a fluorescence with an imaging fluorometer. We compared selected fluorescence parameters (FP) determined from the measurements of control (no stress) and afterwards stressed sample by classical statistical comparison (Mann-Whitney test) and by statistical comparison of shapes of distributions of the FPs (two-sample Smirnov test). We have found that there exist examples where statistically significant difference is not revealed using the classical statistical comparison (for given critical level), but statistically significant difference is revealed using comparisons of distributions (for the same critical level). It implies that the shape of statistical distribution of a FP is more sensitive to a stress of a sample than median of the FP. Further, the comparison of changes in shapes of statistical distributions of FPs is therefore more suitable for early detection of plant stress than a classical statistical comparison. The observed changes in the distributions of FPs are discussed.
Spectral hemispherical reflectance R(lambda) and transmittance T(lambda) are affected by chlorophyll (Chl) fluorescence which may complicate the evaluation of optical parameters of leaves. Measured Chl a fluorescence spectral emission F(lambda) is itself affected by several distortion effects on the leaf level (fluorescence reabsorption, secondary fluorescence, inner filter, surface and subsurface reflections etc.). In this work we propose a Monte Carlo photon transport (MCPT) model capable for treating a variety of optical distortion effects on the leaf level. In the forward mode the model decouples R(lambda), T(lambda) and their fluorescence contributions FR(T)(lambda). To obtain the absorption and scattering spectra of the leaf, utilized in the forward modeling, we have suggested an inversion procedure employing the experimental R(lambda), T(lambda). The attention was paid on the correction of the leaf absorption and scattering spectra caused by the optical effects on the sample level including Chl fluorescence contribution to measured R(lambda), T(lambda).
Commented SBML codes (.XML files) of the monomeric and dimeric PSII models will be available (at the time of publication) in the BioModels database (www.ebi.ac.uk/biomodels).
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