Katrine Heinsvig Kjaer

Smaller stomata require less severe leaf drying to close: A case study in Rosa hydrida

Stomata formed at high relative air humidity (RH) close less as leaf dries; an effect that varies depending on the genotype. We here quantified the contribution of each stomatal response characteristic to the higher water loss of high RH-grown plants, and assessed the relationship between response characteristics and intraspecific variation in stomatal size. Stomatal size (length multiplied by width), density and responsiveness to desiccation, as well as pore dimensions were analyzed in ten rose cultivars grown at moderate (60%) or high (85%) RH. Leaf morphological components and transpiration at growth conditions were also assessed. High growth RH resulted in thinner (11%) leaves with larger area. A strong positive genetic correlation of daytime and nighttime transpiration at either RH was observed. Stomatal size determined pore area (r=0.7) and varied by a factor of two, as a result of proportional changes in length and width. Size and density of stomata were not related. Following desiccation, high RH resulted in a significantly lower (6-19%) decline of transpiration in three cultivars, whereas the relative water content (RWC) of high RH-expanded leaflets was lower (29-297%) in seven cultivars. The lower RWC of these leaflets was caused by (a) higher (33-72%) stable transpiration and/or (b) lower (12-143%) RWC at which this stable transpiration occurred, depending on the cultivar. Stomatal size was significantly correlated with both characteristics (r=0.5 and -0.7, respectively). These results indicate that stomatal size explains much of the intraspecific variation in the regulation of transpiration upon water deprivation on rose.

Threshold response of stomatal closing ability to leaf abscisic acid concentration during growth

Summary Leaf abscisic acid concentration mediates the growth environment-induced effects on both the control of water loss during desiccation and the restoration of water uptake upon re-watering.

Foliar abscisic acid content underlies genotypic variation in stomatal responsiveness after growth at high relative air humidity

BACKGROUND AND AIMS Stomata formed at high relative air humidity (RH) respond less to abscisic acid (ABA), an effect that varies widely between cultivars. This study tested the hypotheses that this genotypic variation in stomatal responsiveness originates from differential impairment in intermediates of the ABA signalling pathway during closure and differences in leaf ABA concentration during growth. METHODS Stomatal anatomical features and stomatal responsiveness to desiccation, feeding with ABA, three transduction elements of its signalling pathway (H2O2, NO, Ca(2+)) and elicitors of these elements were determined in four rose cultivars grown at moderate (60 %) and high (90 %) RH. Leaf ABA concentration was assessed throughout the photoperiod and following mild desiccation (10 % leaf weight loss). KEY RESULTS Stomatal responsiveness to desiccation and ABA feeding was little affected by high RH in two cultivars, whereas it was considerably attenuated in two other cultivars (thus termed sensitive). Leaf ABA concentration was lower in plants grown at high RH, an effect that was more pronounced in the sensitive cultivars. Mild desiccation triggered an increase in leaf ABA concentration and equalized differences between leaves grown at moderate and high RH. High RH impaired stomatal responses to all transduction elements, but cultivar differences were not observed. CONCLUSIONS High RH resulted in decreased leaf ABA concentration during growth as a result of lack of water deficit, since desiccation induced ABA accumulation. Sensitive cultivars underwent a larger decrease in leaf ABA concentration rather than having a higher ABA concentration threshold for inducing stomatal functioning. However, cultivar differences in stomatal closure following ABA feeding were not apparent in response to H2O2 and downstream elements, indicating that signalling events prior to H2O2 generation are involved in the observed genotypic variation.

3D Laser Triangulation for Plant Phenotyping in Challenging Environments.

To increase the understanding of how the plant phenotype is formed by genotype and environmental interactions, simple and robust high-throughput plant phenotyping methods should be developed and considered. This would not only broaden the application range of phenotyping in the plant research community, but also increase the ability for researchers to study plants in their natural environments. By studying plants in their natural environment in high temporal resolution, more knowledge on how multiple stresses interact in defining the plant phenotype could lead to a better understanding of the interaction between plant responses and epigenetic regulation. In the present paper, we evaluate a commercial 3D NIR-laser scanner (PlantEye, Phenospex B.V., Herleen, The Netherlands) to track daily changes in plant growth with high precision in challenging environments. Firstly, we demonstrate that the NIR laser beam of the scanner does not affect plant photosynthetic performance. Secondly, we demonstrate that it is possible to estimate phenotypic variation amongst the growth pattern of ten genotypes of Brassica napus L. (rapeseed), using a simple linear correlation between scanned parameters and destructive growth measurements. Our results demonstrate the high potential of 3D laser triangulation for simple measurements of phenotypic variation in challenging environments and in a high temporal resolution.

Continuous light increases growth, daily carbon gain, antioxidants, and alters carbohydrate metabolism in a cultivated and a wild tomato species

Cultivated tomato species develop leaf injury while grown in continuous light (CL). Growth, photosynthesis, carbohydrate metabolism and antioxidative enzyme activities of a cultivated (Solanum lycopersicum L. ‘Aromata’) and a wild tomato species (Solanum pimpinellifolium L.) were compared in this study aiming to analyze the species-specific differences and thermoperiod effects in responses to CL. The species were subjected to three photoperiodic treatments for 12 days in climate chambers: 16-h photoperiod with a light/dark temperature of 26/16°C (P16D10 or control); CL with a constant temperature of 23°C (P24D0); CL with a variable temperature of 26/16°C (P24D10). The results showed that both species grown in CL had higher dry matter production due to the continuous photosynthesis and a subsequent increase in carbon gain. In S. lycopersicum, the rate of photosynthesis and the maximum photochemical efficiency of photosystem II declined in CL with the development of leaf chlorosis, reduction in the leaf chlorophyll content and a higher activity of antioxidative enzymes. The normal diurnal patterns of starch and sugar were only present under control conditions. The results demonstrated that CL conditions mainly affected the photosynthetic apparatus of a cultivated species (S. lycopersicum), and to a less degree to the wild species (S. pimpinellifolium). The negative effects of the CL could be alleviated by diurnal temperature variations, but the physiological mechanisms behind these are less clear. The results also show that the genetic potential for reducing the negative effects of CL does exist in the tomato germplasm.

Photoperiodic variations induce shifts in the leaf metabolic profile of Chrysanthemum morifolium

Plants have a high ability to adjust their metabolism, growth and development to changes in the light environment and to photoperiodic variation, but the current knowledge on how changes in metabolite contents are associated with growth and development is limited. We investigated the effect of three different photoperiodic treatments with similar daily light integral (DLI) on the growth responses and diurnal patterns in detected leaf metabolites in the short day plant Chrysanthemum×morifolium Ramat. Treatments were long day (LD, 18h light/6h dark), short day (SD, 12h light/12h dark) and short day with irregular night interruptions (NI-SD,12h light/12h dark, applied in a weekly pattern, shifting from day-to-day). Photoperiodic variation resulted in changes in the phenotypic development of the plants. The plants grown in the SD treatment started to initiate reproductive development of the meristems and a decrease in leaf expansion resulted in lower leaf area of expanding leaves. In contrast, plants in the NI-SD and LD treatments did not show reproductive development at any stage and final leaf area of the expanding leaves was intermediate for the NI-SD plants and largest for the LD plants. Photoperiodic variation also resulted in changes in the leaf metabolic profile for most of the analysed metabolites, but only carbohydrates, citrate and some amino acids displayed a shift in their diurnal pattern. Further, our results illustrated that short days (SD) increased the diurnal turnover of 1-kestose after 2 weeks, and decreased the overall contents of leaf hexoses after 3 weeks. In the two other treatments a diurnal turnover of 1-kestose was not stimulated before after 3 weeks, and hexoses together with the hexose:sucrose ratio steadily increased during the experiment. Our results enlighten the plasticity of leaf growth and metabolism to environmental changes, and demonstrate that diurnally regulated metabolites not always respond to photoperiodic variation.

DynaGrow – Multi-Objective Optimization for Energy Cost-efficient Control of Supplemental Light in Greenhouses

The Danish greenhouse horticulture industry utilized 0.8 % of the total national electricity consumption in 2009 and it is estimated that 75 % of this is used for supplemental lighting. The increase in energy prices is a challenge for growers, and need to be addressed by utilizing supplemental light at low prices without compromising the growth and quality of the crop. Optimization of such multiple conflicting objectives requires advanced strategies that are currently not supported in existing greenhouse climate control systems. It is costly to incorporate advanced optimization functionality into existing systems as the software is not designed for such changes. The growers can not afford to buy new systems or new hardware to address the changing objectives. DynaGrow is build on top of existing climate computers to utilize existing infrastructure. The greenhouse climate control problem is characterized by non-linearity , stochasticity, non-convexity, high dimension of decision variables and an uncertain dynamic environment. Together, these mathematical properties are handled by applying a Multi-Objective Evolutionary Algorithm (MOEA) for discovering and exploiting critical trade-offs when optimizing the greenhouse climate. To formulate advanced objectives, DynaGrow integrates local climate data, electricity energy price forecasts and outdoor weather forecasts. In spring 2015, one greenhouse experiment was executed to evaluate the effects of DynaGrow. The experiment was run as three treatments in three identical greenhouse compartments. One treatment was controlled by a standard control system and the other three treatments were controlled by different DynaGrow configurations. A number of different plant species and batches were grown in the three treatments over a season. The results from DynaGrow treatment demonstrated that it was clearly possible to produce a number of different sales-ready plant species and at the same time optimize the utility of supplemental light at low electricity prices without compromising product quality.

Temperature Variation under Continuous Light Restores Tomato Leaf Photosynthesis and Maintains the Diurnal Pattern in Stomatal Conductance

The response of tomato plants (Solanum lycopersicum L. cv. Aromata) to continuous light (CL) in relation to photosynthesis, abscisic acid (ABA) and reactive oxygen species (ROS) was investigated to improve the understanding of the development and/or alleviation of CL-induced leaf injury in constant and diurnal temperature fluctuations with similar daily light integral and daily mean temperature. The plants were grown in three photoperiodic treatments for 15 days; One treatment with a 16/8 h light/dark period and a light/dark temperature of 27/17°C (Control), two CL treatments with 24 h photoperiods, one with a constant temperature of 24°C (CLCT) and the other one with variable temperature of 27/17°C for 16/8 ho, respectively (CLVT). A diurnal pattern of stomatal conductance (gs) and [ABA] was observed in the plants grown in the control and CLVT conditions, while the plants in CLCT conditions experienced a significant decrease in stomatal conductance aligned with an increase in ABA. The net photosynthesis (A) was significantly reduced in CLCT, aligned with a significant decrease in the maximum rate of Rubisco carboxylation (Vcmax), the maximum rate of electron transport (Jmax) and mesophyll diffusion conductance to CO2 (gm) in comparison to the control and CLVT. An increased production of H2O2 and O2•- linked with increased activities of antioxidative enzymes was seen in both CL treatments, but despite of this, leaf injuries were only observed in the CLCT treatment. The results suggest that the diurnal temperature fluctuations alleviated the CL injury symptoms, probably because the diurnal cycles of cellular mechanisms were maintained. The ROS were shown not to be directly involved in CL-induced leaf injury, since both ROS production and scavenging was highest in CLVT without leaf chlorotic symptoms.

DynaGrow: Next Generation Software for Multi-Objective and Energy Cost-Efficient Control of Supplemental Light in Greenhouses

It is not possible for growers to compromise product quality by saving energy but the increasing electricity prices challenge the growers economically. Optimization of such multiple conflicting goals requires advanced strategies that are currently not supported in existing greenhouse climate control systems. DynaGrow is built on top of the existing climate control computers and utilizes the existing hardware. By integrating with exiting hardware it is possibly to support advanced multi-objective optimization of climate parameters without investing in new hardware. Furthermore, DynaGrow integrates with local climate data, electricity price forecasts and outdoor weather forecasts, in order to formulate advanced control objectives. In September 2014 and February 2015 two greenhouse experiments were run to evaluate the effects of DynaGrow. By applying multi-objective optimization, it was possible to produce a number of different cultivars and save energy without compromising quality. The best energy savings were achieved in the February 2015 experiment where the contribution from natural light was limited.

Metabolic fingerprinting of dormant and active flower primordia of Ribes nigrum using HR-MAS NMR

Global warming may modify the timing of dormancy release and spring growth of buds of temperate fruit crops. Environmental regulation of the activity-dormancy cycle in perennial plants remains poorly understood at the metabolic level. Especially, the fine-scale metabolic dynamics in the meristematic zone within buds has received little attention. In this work we performed metabolic profiling of intact floral primordia of Ribes nigrum isolated from buds differing in dormancy status using high-resolution magic angle spinning (HR-MAS) NMR. The technique proved useful in monitoring different groups of metabolites, e.g., carbohydrates and amino acids, in floral primordia and allowed metabolic separation of primordia from endo- and ecodormant buds. In addition, due to its nondestructive character, HR-MAS NMR may provide novel insights into cellular compartmentation of individual biomolecules that cannot be obtained using liquid-state NMR. Out results show that HR-MAS NMR may be an important method for metabolomics of intact plant structures.