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Image-based high-throughput phenotyping promises the rapid determination of functional traits in large plant populations. However, interpretation of some traits – such as those related to photosynthesis or transpiration rates – is only meaningful if the irradiance absorbed by the measured leaves is known, which can differ greatly between different parts of the same plant and within canopies. No feasible method currently exists to rapidly measure absorbed irradiance in three-dimensional plants and canopies. We developed a method and protocols to derive absorbed irradiance at any visible part of a canopy with a thermal camera, by fitting a leaf energy balance model to transient changes in leaf temperature. Leaves were exposed to short light pulses (30 s) that were not long enough to trigger stomatal opening but strong enough to induce transient changes in leaf temperature that was proportional to the absorbed irradiance. The method was successfully validated against point measurements of absorbed irradiance in plant species with relatively simple architecture (sweet pepper, cucumber, tomato, and lettuce). Once calibrated, the model was used to produce absorbed irradiance maps from thermograms. Our method opens new avenues for the interpretation of plant responses derived from imaging techniques and can be adapted to existing high-throughput phenotyping platforms.

To ensure a sustainable future and combat food scarcity, we must boost agricultural productivity, improve climate resilience and optimize resource usage. There is untapped potential for dense wireless sensor networks in agriculture that can increase yields and support resilient production when linked to smart decision and control systems., Green Open Access added to TU Delft Institutional Repository 'You share, we take care!' - Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public., Dynamics of Micro and Nano Systems, Water Resources

Several vital processes in crop canopies and their microclimate, such as canopy photosynthesis and transpiration, are affected by fluctuations in irradiance, here called lightflecks. These lightflecks occur both in the field and inside greenhouses, although their dynamics may be different, due to an absence of wind in the greenhouse. When modelling plant and climate processes, these dynamic conditions are often ignored. The current study reports (1) high frequency (i.e. 1s interval) observations of lightflecks inside a greenhouse during several months and (2) a quantification of the effect of these lightflecks on leaf-level photosynthesis, transpiration and energy balance in rose by comparing results from a dynamic and a steady-state biophysical model. We found that lightflecks in the greenhouse were short, typically <5s. Increasing complexity in irradiance fluctuation patterns (i.e. from very artificial to measured time series) reduced the difference between dynamic and steady-state simulations, e.g. for photosynthesis this difference decreased from 50% to 5%; for water-use efficiency the difference decreased from 70% to 5%. The relative contribution of the increase in light sum and fewer delayed responses by compensating drops in light level through additional light supply on plant processes was different. For example, photosynthesis was equally affected by both the increase in light sum and fewer delayed responses (i.e. 19% and 15% respectively), whereas water-use efficiency was mostly a result of less delayed plant responses (i.e. 9% compared to 1%). Our results demonstrate that lightflecks occur frequently in the greenhouse, and have strong effects on crop photosynthesis, water-use efficiency, and microclimate conditions.

The conversion of supplemental greenhouse light energy into biomass is not always optimal. Recent trends in global energy prices and discussions on climate change highlight the need to reduce our energy footprint associated with the use of supplemental light in greenhouse crop production. This can be achieved by implementing “smart” lighting regimens which in turn rely on a good understanding of how fluctuating light influences photosynthetic physiology. Here, a simple fit-for-purpose dynamic model is presented. It accurately predicts net leaf photosynthesis under natural fluctuating light. It comprises two ordinary differential equations predicting: 1) the total stomatal conductance to CO2 diffusion and 2) the CO2 concentration inside a leaf. It contains elements of the Farquhar-von Caemmerer-Berry model and the successful incorporation of this model suggests that for tomato (Solanum lycopersicum L.), it is sufficient to assume that Rubisco remains activated despite rapid fluctuations in irradiance. Furthermore, predictions of the net photosynthetic rate under both 400ppm and enriched 800ppm ambient CO2 concentrations indicate a strong correlation between the dynamic rate of photosynthesis and the rate of electron transport. Finally, we are able to indicate whether dynamic photosynthesis is Rubisco or electron transport rate limited.

Plant breeding for increased crop water use efficiency or drought stress resistance requires methods to quickly assess the transpiration rate (E) and stomatal conductance (gs) of a large number of individual plants. Several methods to measure E and gs exist, each of which has its own advantages and shortcomings. To add to this toolbox, we developed a method that uses whole-plant thermal imaging in a controlled environment, where aerial humidity is changed rapidly to induce changes in E that are reflected in changes in leaf temperature. This approach is based on a simplified energy balance equation, without the need for a reference material or complicated calculations. To test this concept, we built a double-sided, perforated, open-top plexiglass chamber that was supplied with air at a high flow rate (35 L min−1) and whose relative humidity (RH) could be switched rapidly. Measurements included air and leaf temperature as well as RH. Using several well-watered and drought stressed genotypes of Arabidopsis thaliana that were exposed to multiple cycles in RH (30–50 % and back), we showed that leaf temperature as measured in our system correlated well with E and gs measured in a commercial gas exchange system. Our results demonstrate that, at least within a given species, the differences in leaf temperature under several RH can be used as a proxy for E and gs. Given that this method is fairly quick, noninvasive and remote, we envision that it could be upscaled for work within rapid plant phenotyping systems.

Salinity is a current and growing problem, affecting crops worldwide by reducing yields and product quality. Plants have different mechanisms to adapt to salinity; some crops are highly studied, and their salinity tolerance mechanisms are widely known. However, there are other crops with commercial importance that still need characterization of their molecular mechanisms. Usually, transcription factors are in charge of the regulation of complex processes such as the response to salinity. MYB-TFs are a family of transcription factors that regulate various processes in plant development, and both central and specialized metabolism. MYB-TFs have been studied extensively as mediators of specialized metabolism, and some are master regulators. The influence of MYB-TFs on highly orchestrated mechanisms, such as salinity tolerance, is an attractive research target. The versatility of petunia as a model species has allowed for advances to be made in multiple fields: metabolomic pathways, quality traits, stress resistance, and signal transduction. It has the potential to be the link between horticultural crops and lab models, making it useful in translating discoveries related to the MYB-TF pathways into other crops. We present a phylogenetic tree made with Petunia axillaris and Petunia inflata R2R3-MYB subfamily sequences, which could be used to find functional conservation between different species. This work could set the foundations to improve salinity resistance in other commercial crops in later studies.

NaCl stress affects stomatal behavior and photosynthesis by a combination of osmotic and ionic components, but it is unknown how these components affect stomatal and photosynthetic dynamics. Tomato (Solanum lycopersicum) plants were grown in a reference nutrient solution [control; electrical conductivity (EC)=2.3 dS m–1], a solution containing additional macronutrients (osmotic effect; EC=12.6 dS m–1), or a solution with additional 100 mM NaCl (osmotic and ionic effects; EC=12.8 dS m–1). Steady-state and dynamic photosynthesis, and leaf biochemistry, were characterized throughout leaf development. The osmotic effect decreased steady-state stomatal conductance while speeding up stomatal responses to light intensity shifts. After 19 d of treatment, photosynthetic induction was reduced by the osmotic effect, which was attributable to lower initial stomatal conductance due to faster stomatal closing under low light. Ionic effects of NaCl were barely observed in dynamic stomatal and photosynthetic behavior, but led to a reduction in leaf photosynthetic capacity, CO2 carboxylation rate, and stomatal conductance in old leaves after 26 d of treatment. With increasing leaf age, rates of light-triggered stomatal movement and photosynthetic induction decreased across treatments. We conclude that NaCl impacts dynamic stomatal and photosynthetic kinetics by osmotic effects and reduces photosynthetic capacity by ionic effects.

Plant transpiration accounts for about half of all terrestrial evaporation. Plants need water for many vital functions including nutrient uptake, growth and leaf cooling. The regulation of plant water transport by stomata in the leaves leads to the loss of 97% of the water that is taken up via their roots, to the atmosphere. Measuring plant-water dynamics is essential to gain better insight into its roles in the terrestrial water cycle and plant productivity. It can be measured at different levels of integration, from the single cell micro-scale to the ecosystem macro-scale, on time scales from minutes to months. In this contribution, we give an overview of state-of-the-art techniques for plant-water dynamics measurement and highlight several promising innovations for future monitoring. Some of the techniques we will cover include: gas exchange for stomatal conductance and transpiration monitoring, lysimetry, thermometry, heat-based sap flow monitoring, reflectance monitoring including satellite remote sensing, ultrasound spectroscopy, dendrometry, accelometry, scintillometry, stable water isotope analysis and eddy covariance. To fully assess water transport within the soil-plant-atmosphere continuum, a variety of techniques are required to monitor environmental variables in combination with biological responses at different scales. Yet this is not sufficient: to truly account for spatial heterogeneity, a dense network sampling is needed., Green Open Access added to TU Delft Institutional Repository ‘You share, we take care!’ – Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public., Dynamics of Micro and Nano Systems, Water Resources

Although it is well known that plants emit acoustic pulses under drought stress, the exact origin of the waveform of these ultrasound pulses has remained elusive. Here, we present evidence for a correlation between the characteristics of the waveform of these pulses and the dimensions of xylem conduits in plants. Using a model that relates the resonant vibrations of a vessel to its dimension and viscoelasticity, we extract the xylem radii from the waveforms of ultrasound pulses and show that these are correlated and in good agreement with optical microscopy. We demonstrate the versatility of the method by applying it to shoots of ten different vascular plant species. In particular, for Hydrangea quercifolia, we further extract vessel element lengths with our model and compare them with scanning electron cryomicroscopy. The ultrasonic, noninvasive characterization of internal conduit dimensions enables a breakthrough in speed and accuracy in plant phenotyping and stress detection., Dynamics of Micro and Nano Systems, QN/Steeneken Lab

Although it is well known that plants emit acoustic pulses under drought stress, the exact origin of the waveform of these ultrasound pulses has remained elusive. Here, we present evidence for a correlation between the characteristics of the waveform of these pulses and the dimensions of xylem conduits in plants. Using a model that relates the resonant vibrations of a vessel to its dimension and viscoelasticity, we extract the xylem radii from the waveforms of ultrasound pulses and show that these are correlated and in good agreement with optical microscopy. We demonstrate the versatility of the method by applying it to shoots of ten different vascular plant species. In particular, for Hydrangea quercifolia, we further extract vessel element lengths with our model and compare them with scanning electron cryomicroscopy. The ultrasonic, noninvasive characterization of internal conduit dimensions enables a breakthrough in speed and accuracy in plant phenotyping and stress detection.

Raising young plants is important for modern greenhouse production. Upon transfer from the raising to the production environment, young plants should maximize light use efficiency while minimizing deleterious effects associated with exposure to high light (HL) intensity. The light spectrum may be used to establish desired traits, but how plants acclimated to a given spectrum respond to HL intensity exposure is less well explored. Cucumber (Cucumis sativus) seedlings were grown in a greenhouse in low-intensity sunlight (control; ∼2.7 mol photons m–2 day–1) and were treated with white, red, blue, or green supplemental light (4.3 mol photons m–2 day–1) for 10 days. Photosynthetic capacity was highest in leaves treated with blue light, followed by white, red, and green, and was positively correlated with leaf thickness, nitrogen, and chlorophyll concentration. Acclimation to different spectra did not affect the rate of photosynthetic induction, but leaves grown under blue light showed faster induction and relaxation of non-photochemical quenching (NPQ) under alternating HL and LL intensity. Blue-light-acclimated leaves showed reduced photoinhibition after HL intensity exposure, as indicated by a high maximum quantum yield of photosystem II photochemistry (Fv/Fm). Although plants grown under different supplemental light spectra for 10 days had similar shoot biomass, blue-light-grown plants (B-grown plants) showed a more compact morphology with smaller leaf areas and shorter stems. However, after subsequent, week-long exposure to full sunlight (10.7 mol photons m–2 day–1), B-grown plants showed similar leaf area and 15% higher shoot biomass, compared to plants that had been acclimated to other spectra. The faster growth rate in blue-light-acclimated plants compared to other plants was mainly due to a higher photosynthetic capacity and highly regulated NPQ performance under intermittent high solar light. Acclimation to blue supplemental light can improve light use efficien

Using light emitting diodes (LED) instead of conventionally used high pressure sodium (HPS) lamps as a supplemental light source in greenhouses results in a higher efficacy (µmol light per J electricity) and makes it possible to customize the light spectrum. To explore the effects of LED and HPS on gas exchange, thermal relations, photosynthesis, and water status of young tomato plants, seven genotypes were grown in a greenhouse under LED (95% red, 5% blue) or HPS lamps in four experiments differing in the fraction of lamp light over natural light. HPS lights emit a broader spectrum of red (40%), green–yellow (50%), blue (5%), and far-red (5%) and a substantial amount of infrared radiation (heat). Young tomato plants grown under LED showed lower leaf temperature and higher stomatal density, stomatal conductance (gs ) and transpiration rate (E) than plants grown under HPS; this may be due to the different supplemental light spectrum. The young plants grown under LED tended to have increased photosynthetic capacity. Furthermore, the water stress indices CWSI and IG, which were obtained using thermal imaging, were positively correlated with gas exchange-derived gs and E, putting forward the use of thermal imaging for the phenotyping of transpiration. Under LED light, photosynthetic gas exchange was generally increased, which agreed with the water stress indices. The extent of this increase was genotype-dependent. All differences between LED and HPS were smaller in the experiments where the fraction of lamp light over natural light was smaller.

The capacity of photoautotrophs to fix carbon depends on the efficiency of the conversion of light energy into chemical potential by photosynthesis. In nature, light input into photosynthesis can change very rapidly and dramatically. To analyze how genetic variation in Arabidopsis thaliana affects photosynthesis and growth under dynamic light conditions, 36 randomly chosen natural accessions were grown under uniform and fluctuating light intensities. After 14 days of growth under uniform or fluctuating light regimes, maximum photosystem II quantum efficiency (Fv/Fm) was determined, photosystem II operating efficiency (ΦPSII) and non‐photochemical quenching (NPQ) were measured in low light, and projected leaf area (PLA) as well as the number of visible leaves were estimated. Our data show that ΦPSII and PLA were decreased and NPQ was increased, while Fv/Fm and number of visible leaves were unaffected, in most accessions grown under fluctuating compared to uniform light. There were large changes between accessions for most of these parameters, which, however, were not correlated with genomic variation. Fast growing accessions under uniform light showed the largest growth reductions under fluctuating light, which correlated strongly with a reduction in ΦPSII, suggesting that, under fluctuating light, photosynthesis controls growth and not vice versa.

Due to their slow movement and closure upon shade, partially closed stomata can be a substantial limitation to photosynthesis in variable light intensities. The abscisic acid deficient flacca mutant in tomato (Solanum lycopersicum) displays very high stomatal conductance (gs). We aimed to determine to what extent this substantially increased gs affects the rate of photosynthetic induction. Steady-state and dynamic photosynthesis characteristics were measured in flacca and wildtype leaves, by the use of simultaneous gas exchange and chlorophyll fluorometry. The steady-state response of photosynthesis to CO2, maximum quantum efficiency of photosystem II photochemistry (Fv/Fm), as well as mesophyll conductance to CO2 diffusion were not significantly different between genotypes, suggesting similar photosynthetic biochemistry, photoprotective capacity, and internal CO2 permeability. When leaves adapted to shade (50 µmol m−2 s−1) at 400 µbar CO2 partial pressure and high humidity (7 mbar leaf-to-air vapour pressure deficit, VPD) were exposed to high irradiance (1500 µmol m−2 s−1), photosynthetic induction was faster in flacca compared to wildtype leaves, and this was attributable to high initial gs in flacca (~0.6 mol m−2 s−1): in flacca, the times to reach 50 (t50) and 90% (t90) of full photosynthetic induction were 91 and 46% of wildtype values, respectively. Low humidity (15 mbar VPD) reduced gs and slowed down photosynthetic induction in the wildtype, while no change was observed in flacca; under low humidity, t50 was 63% and t90 was 36% of wildtype levels in flacca. Photosynthetic induction in low CO2 partial pressure (200 µbar) increased gs in the wildtype (but not in flacca), and revealed no differences in the rate of photosynthetic induction between genotypes. Effects of higher gs in flacca were also visible in transients of photosystem II operating efficiency and non-photochemical quenching. Our results show that at ambient CO2 partial pressure, wildtype gs is a

Wageningen University & Research, Business Unit Glastuinbouw has investigated two methods of monitoring crop photosynthesis that might be used in greenhouse horticulture: the crop photosynthesis monitor and the CropObserver. This project was funded by ‘Kas als Energiebron’ and executed together with PhenoVation and PlantDynamics. The data of the crop photosynthesis monitor were compared to those of the photosynthesis module of the crop growth model INTKAM. Furthermore, the pattern of stomatal opening during the day was calculated with the Stomata sensor. Measurements showed that the crop photosynthesis monitor was too sensitive to changes in window opening and in CO2 supply to determine the crop photosynthesis accurately. The CropObserver was useful in determining the relative light use efficiency of the crop. The best methods to accurately and reliably determine crop photosynthesis might be the following: 1. By a photosynthesis model (as in INTKAM) to calculate the crop photosynthesis, coupled to a sensor that monitors the crop status to register when the performance of the crop deviates from what might be expected, or 2. Via the CropObserver to which a module is added that determines the stomatal conductance and a measurement of the CO2 concentration which is necessary to translate the signal of the CropObserver to CO2 uptake by the crop, Wageningen University & Research, Business Unit Glastuinbouw heeft samen met PhenoVation en PlantDynamics in opdracht van ‘Kas als Energiebron’ twee methodes om de gewasfotosynthese te monitoren onderzocht: de gewasfotosynthesemonitor en de CropObserver. De data van de gewasfotosynthesemonitor werden vergeleken met de fotosynthesemodule van het gewasgroeimodel INTKAM. Verder werd het dagverloop van de huidmondjesopening bepaald met de Stomatasensor. Uit de metingen bleek dat de gewasfotosynthesemonitor te gevoelig is voor veranderingen in raamstanden en wisselingen in CO2 dosering om de gewasfotosynthese nauwkeurig te kunnen bepalen. De CropObserver bleek een goed beeld te geven van de relatieve lichtbenutting door het gewas. De beste manieren om de gewasfotosynthese nauwkeurig en betrouwbaar te bepalen, zouden de volgende kunnen zijn: 1. Via een fotosynthesemodelmodel (zoals INTKAM) waarmee de gewasfotosynthese berekend kan worden, gekoppeld aan een sensor die de status van het gewas monitort zodat geregistreerd wordt wanneer de reactie van het gewas afwijkt van wat te verwachten zou zijn, of 2. Via de CropObserver, aangevuld met een sensor voor de huidmondjesopening en een meting van tenminste de CO2 concentratie, waarmee het signaal van de CropObserver vertaald kan worden naar CO2 opname door het gewas

A better understanding of the metabolic and diffusional limitations of photosynthesis in fluctuating irradiance can help identify targets for improving crop yields. We used different genotypes of Arabidopsis thaliana to characterise the importance of Rubisco activase (Rca), stomatal conductance (g s), non-photochemical quenching of chlorophyll fluorescence (NPQ) and sucrose phosphate synthase (SPS) on photosynthesis in fluctuating irradiance. Leaf gas exchange and chlorophyll fluorescence were measured in leaves exposed to stepwise increases and decreases in irradiance. rwt43, which has a constitutively active Rubisco enzyme in different irradiance intensities (except in darkness), showed faster increases than the wildtype, Colombia-0, in photosynthesis rates after step increases in irradiance. rca-2, having decreased Rca concentration, showed lower rates of increase. In aba2-1, high g s increased the rate of change after stepwise irradiance increases, while in C24, low gs tended to decrease it. Differences in rates of change between Colombia-0 and plants with low levels of NPQ (npq1-2, npq4-1) or SPS (spsa1) were negligible. In Colombia-0, the regulation of Rubisco activation and of gs were therefore limiting for photosynthesis in fluctuating irradiance, while levels of NPQ or SPS were not. This suggests Rca and g s as targets for improvement of photosynthesis of plants in fluctuating irradiance.