7 research outputs found

    Passive direct measurement of sun-induced chlorophyll fluorescence spectrum from in vivo leaves

    Get PDF
    The fluorescence of chlorophyll in vegetation is a weak signal emitted between 650 and 850 nm that is mixed with the much more intense light reflected by the leaf, which is why active methods are commonly used (through the additional contribution of controlled artificial light) or using indirect measurements instead. So, the measurement is provided just in relative units in the first case, or the accuracy of the estimate in the second case is uncertain without proper direct validation. The Thesis presents a new device, called FluoWat, for passive measurement that allows direct measurement of the fluorescence emission of leaves in vivo under natural conditions in the field with sunlight. And it is part of the activities supporting the preparation of ESA’s FLEX mission for the global monitoring of vegetation fluorescence. The device consists of a small dark chamber implemented as a clip, so that the leaf can be housed inside without damaging it, with an opening to illuminate the sample by pointing at the sun, and a sliding filter holder with a low-pass filter that blocks sunlight in the same spectral range as fluorescence is emitted while allowing the excitation light to pass through, then a spectroradiometer connected to the clip measures the fluorescence spectrum without interference from sunlight. In addition, it is possible to measure the reflectance and transmittance factors of the leaf, which allows determining the absorptance, necessary to determine the photosynthetically active radiation (PAR) that has been absorbed (APAR). An essential parameter to properly interpret the fluorescence signal in relation to photosynthesis. Similarly, the reflectance and transmittance spectra in the visible range make it possible to determine the degree of photoprotection of the leaf and/or its chlorophyll content. A sensitivity analysis of different factors likely to affect the measurement has been carried out, such as the residual light that passes through the filter, or the effect of transients on fluorescence emission, among others. Processing methods have been developed to mitigate their effects on the fluorescence measurement, increasing the accuracy of the results. Finally, a series of experiments are presented in which the system is put to the test and that illustrate how, with the measurements provided by this new device, a better understanding of the dynamics of fluorescence emission while the vegetation adapts to different illumination changes, levels of stress and changing environmental conditions

    In vivo photoprotection mechanisms observed from leaf spectral absorbance changes showing VIS-NIR slow-induced conformational pigment bed changes

    Get PDF
    Regulated heat dissipation under excessive light comprises a complexity of mechanisms, whereby the supramolecular light-harvesting pigment-protein complex (LHC) shifts state from light harvesting towards heat dissipation, quenching the excess of photo-induced excitation energy in a non-photochemical way. Based on whole-leaf spectroscopy measuring upward and downward spectral radiance fluxes, we studied spectrally contiguous (hyperspectral) transient time series of absorbance A(λ,t) and passively induced chlorophyll fluorescence F(λ,t) dynamics of intact leaves in the visible and near-infrared wavelengths (VIS-NIR, 400-800 nm) after sudden strong natural-like illumination exposure. Besides light avoidance mechanism, we observed on absorbance signatures, calculated from simultaneous reflectance R(λ,t) and transmittance T(λ,t) measurements as A(λ,t) = 1 − R(λ,t) − T(λ,t), major dynamic events with specific onsets and kinetical behaviour. A consistent well-known fast carotenoid absorbance feature (500-570 nm) appears within the first seconds to minutes, seen from both the reflected (backscattered) and transmitted (forward scattered) radiance differences. Simultaneous fast Chl features are observed, either as an increased or decreased scattering behaviour during quick light adjustment consistent with re-organizations of the membrane. The carotenoid absorbance feature shows up simultaneously with a major F decrease and corresponds to the xanthophyll conversion, as quick response to the proton gradient build-up. After xanthophyll conversion (t = 3 min), a kinetically slower but major and smooth absorbance increase was occasionally observed from the transmitted radiance measurements as wide peaks in the green (~ 550 nm) and the near-infrared (~ 750 nm) wavelengths, involving no further F quenching. Surprisingly, in relation to the response to high light, this broad and consistent VIS-NIR feature indicates a slowly induced absorbance increase with a sigmoid kinetical behaviour. In analogy to sub-leaf-level observations, we suggest that this mechanism can be explained by a structure-induced low-energy-shifted energy redistribution involving both Car and Chl. These findings might pave the way towards a further non-invasive spectral investigation of antenna conformations and their relations with energy quenching at the intact leaf level, which is, in combination with F measurements, of a high importance for assessing plant photosynthesis in vivo and in addition from remote observations

    Photoprotection dynamics observed at leaf level from fast temporal reflectance changes

    Get PDF
    Vegetation dynamically reacts to the available photosynthetically active radiation (PAR) by adjusting the photosynthetic apparatus to either a light harvesting or a photoprotective modus. When activating the photoprotection mechanism, either minor or major pigment-protein interactions may occur at the leaf level, resulting in different light absorption and consequently reflectance intensities. The reflectance changes were measured during sudden illumination transients designed to provoke fast adaptation to high irradiance. Different spectral reflectance change features were observed during different stages of photoprotection activation, extending over part of the visible spectral range (i.e. 490-650 nm). Due to this multiple wavelength reflectance modification, which affects also the reference band at 570 nm, the commonly used Photochemical Reflectance Index (PRI) is unable to trace and quantify such strong photoprotection mechanism. To quantify the entire photoprotection with a required accuracy, the spectral changes in the full visible range must be characterized

    Impact of Structural, Photochemical and Instrumental Effects on Leaf and Canopy Reflectance Variability in the 500-600 nm Range

    Get PDF
    Current rapid technological improvement in optical radiometric instrumentation provides an opportunity to develop innovative measurements protocols where the remote quantification of the plant physiological status can be determined with higher accuracy. In this study, the leaf and canopy reflectance variability in the PRI spectral region (i.e., 500-600 nm) is quantified using different laboratory protocols that consider both instrumental and experimental set-up aspects, as well as canopy structural effects and vegetation photoprotection dynamics. First, we studied how an incorrect characterization of the at-target incoming radiance translated into an erroneous vegetation reflectance spectrum and consequently in an incorrect quantification of reflectance indices such as PRI. The erroneous characterization of the at-target incoming radiance translated into a 2% overestimation and a 31% underestimation of estimated chlorophyll content and PRI-related vegetation indexes, respectively. Second, we investigated the dynamic xanthophyll pool and intrinsic Chl vs. Car long-term pool changes affecting the entire 500-600 nm spectral region. Consistent spectral behaviors were observed for leaf and canopy experiments. Sun-adapted plants showed a larger optical change in the PRI range and a higher capacity for photoprotection during the light transient time when compared to shade-adapted plants. Outcomes of this work highlight the importance of well-established spectroscopy sampling protocols to detect the subtle photochemical features which need to be disentangled from the structural and biological effects

    Towards the quantitative and physically-based interpretation of solar-induced vegetation fluorescence retrieved from global imaging

    Get PDF
    Due to emerging high spectral resolution, remote sensing techniques and ongoing developments to retrieve the spectrally resolved vegetation fluorescence spectrum from several scales, the light reactions of photosynthesis are receiving a boost of attention for the monitoring of the Earth's carbon balance. Sensor-retrieved vegetation fluorescence (from leaf, tower, airborne or satellite scale) originating from the excited antenna chlorophyll a molecule has become a new quantitative biophysical vegetation parameter retrievable from space using global imaging techniques. However, to retrieve the actual quantum efficiencies, and hence a true photosynthetic status of the observed vegetation, all signal distortions must be accounted for, and a high-precision true vegetation reflectance must be resolved. ESA's upcoming Fluorescence Explorer aims to deliver such novel products thanks to technological and instrumental advances, and by sophisticated approaches that will enable a deeper understanding of the mechanics of energy transfer underlying the photosynthetic process in plant canopies and ecosystems
    corecore