Active and passive chlorophyll fluorescence measurements at canopy level on potato crops. Evidence of similitude of diurnal cycles of apparent fluorescence yields.

We performed active and passive measurements of diurnal cycles of chlorophyll fluorescence on potato crops at canopy level in outdoors conditions for 26 days. Active measurements of the stationary fluorescence yield (Fs) were performed using Ledflex, a fluorescence micro-LIDAR described in Moya et al. (Photosynth Res 142:1-15, 2019), capable of remote measurements of chlorophyll fluorescence under full sun-light in the wavelength range from 650 to 800 nm. Passive measurements of solar-induced fluorescence (SIF) fluxes were performed with Spectroflex, an instrument based on the method of filling-in in the O2A and O2B absorption bands at 760 nm (F760) and 687 nm (F687), respectively.Diurnal cycles of Fs showed significant variations throughout the day, directly attributed to changes in photosystem II yield. Contrasting patterns were observed according to illumination conditions. Under cloudy sky, Fs varied in parallel with photosynthetically active radiation (PAR). By contrast, during clear sky days, the diurnal cycle of Fs showed a "M" shape pattern with a minimum around noon.F687 and F760 showed different patterns, according to illumination conditions. Under low irradiance associated with cloudy conditions, F687 and F760 followed similar diurnal patterns, in parallel with PAR. Under high irradiance associated with clear sky we observed an increase of the F760/F687 ratio, which we attributed to the contributions in the 760 nm emission of photosystem I fluorescence from deeper layers of the leaves, on one end, and by the decrease of 687 nm emission as a result of red fluorescence re-absorption, on the other end.We defined an approach to derive a proxy of fluorescence yield (FYSIF) from SIF measurements as a linear combination of F687 and F760 normalized by vegetation radiance, where the coefficients of the linear combination were derived from the spectral transmittance of Ledflex. We demonstrated a close relationship between diurnal cycles of FYSIF and Fs, which outperformed other approaches based on normalization by incident light.

Active in situ and passive airborne fluorescence measurements for water stress detection on a fescue field.

Ledflex is a fluorometer adapted to measure chlorophyll fluorescence at the canopy level. It has been described in detail by Moya et al. (2019), Photosynthesis Research. https://doi.org/10.1007/s11120-019-00642-9 . We used this instrument to determine the effect of water stress on the fluorescence of a fescue field under extreme temperature and light conditions through a 12 days campaign during summer in a Mediterranean area. The fescue field formed part of a lysimeter station in "las Tiesas," near Albacete-Spain. In addition to the fluorescence data, the surface temperature was measured using infrared radiometers. Furthermore, "Airflex," a passive fluorometer measuring the filling-in of the atmospheric oxygen absorption band at 760 nm, was installed in an ultralight plane and flown during the most critical days of the campaign. We observed with the Ledflex fluorometer a considerable decrease of about 53% of the stationary chlorophyll fluorescence level at noon under water stress, which was well correlated with the surface temperature difference between the stressed and control plots. Airflex data also showed a decrease in far-red solar-induced fluorescence upon water stress in agreement with surface temperature data and active fluorescence measurements after correction for PS I contribution. Notwithstanding, the results from airborne remote sensing are not as precise as in situ active data.

Canopy chlorophyll fluorescence applied to stress detection using an easy-to-build micro-lidar.

LEDFLEX is a micro-lidar dedicated to the measurement of vegetation fluorescence. The light source consists of 4 blue Light-Emitting Diodes (LED) to illuminate part of the canopy in order to average the spatial variability of small crops. The fluorescence emitted in response to a 5-µs width pulse is separated from the ambient light through a synchronized detection. Both the reflectance and the fluorescence of the target are acquired simultaneously in exactly the same field of view, as well as the photosynthetic active radiation and air temperature. The footprint is about 1 m2 at a distance of 8 m. By increasing the number of LEDs longer ranges can be reached. The micro-lidar has been successfully applied under full sunlight conditions to establish the signature of water stress on pea (Pisum Sativum) canopy. Under well-watered conditions the diurnal cycle presents an M shape with a minimum (Fmin) at noon which is Fmin > Fo. After several days withholding watering, Fs decreases and Fmin < Fo. The same patterns were observed on mint (Menta Spicata) and sweet potatoes (Ipomoea batatas) canopies. Active fluorescence measurements with LEDFLEX produced robust fluorescence yield data as a result of the constancy of the excitation intensity and its geometry fixity. Passive methods based on Sun-Induced chlorophyll Fluorescence (SIF) that uses high-resolution spectrometers generate only flux data and are dependent on both the 3D structure of vegetation and variable irradiance conditions along the day. Parallel measurements with LEDFLEX should greatly improve the interpretation of SIF changes.

Remote sensing of solar-induced chlorophyll fluorescence (SIF) in vegetation: 50 years of progress.

Remote sensing of solar-induced chlorophyll fluorescence (SIF) is a rapidly advancing front in terrestrial vegetation science, with emerging capability in space-based methodologies and diverse application prospects. Although remote sensing of SIF - especially from space - is seen as a contemporary new specialty for terrestrial plants, it is founded upon a multi-decadal history of research, applications, and sensor developments in active and passive sensing of chlorophyll fluorescence. Current technical capabilities allow SIF to be measured across a range of biological, spatial, and temporal scales. As an optical signal, SIF may be assessed remotely using highly-resolved spectral sensors and state-of-the-art algorithms to distinguish the emission from reflected and/or scattered ambient light. Because the red to far-red SIF emission is detectable non-invasively, it may be sampled repeatedly to acquire spatio-temporally explicit information about photosynthetic light responses and steady-state behaviour in vegetation. Progress in this field is accelerating with innovative sensor developments, retrieval methods, and modelling advances. This review distills the historical and current developments spanning the last several decades. It highlights SIF heritage and complementarity within the broader field of fluorescence science, the maturation of physiological and radiative transfer modelling, SIF signal retrieval strategies, techniques for field and airborne sensing, advances in satellite-based systems, and applications of these capabilities in evaluation of photosynthesis and stress effects. Progress, challenges, and future directions are considered for this unique avenue of remote sensing.

Chlorophyll fluorescence emission spectrum inside a leaf.

Chlorophyll a fluorescence can be used as an early stress indicator. Fluorescence is also connected to photosynthesis so it can be proposed for global monitoring of vegetation status from a satellite platform. Nevertheless, the correct interpretation of fluorescence requires accurate physical models. The spectral shape of the leaf fluorescence free of any re-absorption effect plays a key role in the models and is difficult to measure. We present a vegetation fluorescence emission spectrum free of re-absorption based on a combination of measurements and modelling. The suggested spectrum takes into account the photosystem I and II spectra and their relative contribution to fluorescence. This emission spectrum is applicable to describe vegetation fluorescence in biospectroscopy and remote sensing.

Dualex: a new instrument for field measurements of epidermal ultraviolet absorbance by chlorophyll fluorescence.

Dualex (dual excitation) is a field-portable instrument, hereby described, for the assessment of polyphenolic compounds in leaves from the measurement of UV absorbance of the leaf epidermis by double excitation of chlorophyll fluorescence. The instrument takes advantage of a feedback loop that equalizes the fluorescence level induced by a reference red light to the UV-light-induced fluorescence level. This allows quick measurement from attached leaves even under field conditions. The use of light-emitting diodes and of a leaf-clip configuration makes Dualex a user-friendly instrument with potential applications in ecophysiological research, light climate analysis, agriculture, forestry, horticulture, pest management, selection of medicinal plants, and wherever accumulation of leaf polyphenolics is involved in plant responses to the environment.