Plant Physiological Analysis to Overcome Limitations to Plant Phenotyping.

Plant physiological status is the interaction between the plant genome and the prevailing growth conditions. Accurate characterization of plant physiology is, therefore, fundamental to effective plant phenotyping studies; particularly those focused on identifying traits associated with improved yield, lower input requirements, and climate resilience. Here, we outline the approaches used to assess plant physiology and how these techniques of direct empirical observations of processes such as photosynthetic CO2 assimilation, stomatal conductance, photosystem II electron transport, or the effectiveness of protective energy dissipation mechanisms are unsuited to high-throughput phenotyping applications. Novel optical sensors, remote/proximal sensing (multi- and hyperspectral reflectance, infrared thermography, sun-induced fluorescence), LiDAR, and automated analyses of below-ground development offer the possibility to infer plant physiological status and growth. However, there are limitations to such 'indirect' approaches to gauging plant physiology. These methodologies that are appropriate for the rapid high temporal screening of a number of crop varieties over a wide spatial scale do still require 'calibration' or 'validation' with direct empirical measurement of plant physiological status. The use of deep-learning and artificial intelligence approaches may enable the effective synthesis of large multivariate datasets to more accurately quantify physiological characters rapidly in high numbers of replicate plants. Advances in automated data collection and subsequent data processing represent an opportunity for plant phenotyping efforts to fully integrate fundamental physiological data into vital efforts to ensure food and agro-economic sustainability.

Lipid Peroxidation and Chlorophyll Fluorescence of Photosystem II Performance during Drought and Heat Stress is Associated with the Antioxidant Capacities of C3 Sunflower and C4 Maize Varieties.

Agricultural production is predicted to be adversely affected by an increase in drought and heatwaves. Drought and heat damage cellular membranes, such as the thylakoid membranes where photosystem II occurs (PSII). We investigated the chlorophyll fluorescence (ChlF) of PSII, photosynthetic pigments, membrane damage, and the activity of protective antioxidants in drought-tolerant and -sensitive varieties of C3 sunflower and C4 maize grown at 20/25 and 30/35 °C. Drought-tolerant varieties retained PSII electron transport at lower levels of water availability at both temperatures. Drought and heat stress, in combination and isolation, had a more pronounced effect on the ChlF of the C3 species. For phenotyping, the maximum fluorescence was the most effective ChlF measure in characterizing varietal variation in the response of both species to drought and heat. The drought-tolerant sunflower and maize showed lower lipid peroxidation under drought and heat stress. The greater retention of PSII function in the drought-tolerant sunflower and maize at higher temperatures was associated with an increase in the activities of antioxidants (glutathione reductase, superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase), whereas antioxidant activity declined in the drought-sensitive varieties. Antioxidant activity should play a key role in the development of drought- and heat-tolerant crops for future food security.

The Impact of Heat Stress and Water Deficit on the Photosynthetic and Stomatal Physiology of Olive (Olea europaea L.)-A Case Study of the 2017 Heat Wave.

Heat waves are predicted to increase in frequency and duration in many regions as global temperatures rise. These transient increases in temperature above normal average values will have pronounced impacts upon the photosynthetic and stomatal physiology of plants. During the summer of 2017, much of the Mediterranean experienced a severe heat wave. Here, we report photosynthetic leaf gas exchange and chlorophyll fluorescence parameters of olive (Olea europaea cv. Leccino) grown under water deficit and full irrigation over the course of the heat wave as midday temperatures rose over 40 °C in Central Italy. Heat stress induced a decline in the photosynthetic capacity of the olives consistent with reduced ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) activity. Damage to photosystem II was more apparent in plants subject to water deficit. In contrast to previous studies, higher temperatures induced reductions in stomatal conductance. Heat stress adversely affected the carbon efficiency of olive. The selection of olive varieties with enhanced tolerance to heat stress and/or strategies to mitigate the impact of higher temperatures will become increasingly important in developing sustainable agriculture in the Mediterranean as global temperatures rise.

Publisher Correction: Impaired photosynthesis and increased leaf construction costs may induce floral stress during episodes of global warming over macroevolutionary timescales.

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Impaired photosynthesis and increased leaf construction costs may induce floral stress during episodes of global warming over macroevolutionary timescales.

Global warming events have coincided with turnover of plant species at intervals in Earth history. As mean global temperatures rise, the number, frequency and duration of heat-waves will increase. Ginkgo biloba was grown under controlled climatic conditions at two different day/night temperature regimes (25/20 °C and 35/30 °C) to investigate the impact of heat stress. Photosynthetic CO2-uptake and electron transport were reduced at the higher temperature, while rates of respiration were greater; suggesting that the carbon balance of the leaves was adversely affected. Stomatal conductance and the potential for evaporative cooling of the leaves was reduced at the higher temperature. Furthermore, the capacity of the leaves to dissipate excess energy was also reduced at 35/30 °C, indicating that photo-protective mechanisms were no longer functioning effectively. Leaf economics were adversely affected by heat stress, exhibiting an increase in leaf mass per area and leaf construction costs. This may be consistent with the selective pressures experienced by fossil Ginkgoales during intervals of global warming such as the Triassic - Jurassic boundary or Early Eocene Climatic Optimum. The physiological and morphological responses of the G. biloba leaves were closely interrelated; these relationships may be used to infer the leaf economics and photosynthetic/stress physiology of fossil plants.

Diffusive and Metabolic Constraints to Photosynthesis in Quinoa during Drought and Salt Stress.

Quinoa (Chenopodium quinoa Willd.) has been proposed as a hardy alternative to traditional grain crops in areas with warm-to-hot climates that are likely to experience increased drought and salt stress in the future. We characterised the diffusive and metabolic limitations to photosynthesis in quinoa exposed to drought and salt stress in isolation and combination. Drought-induced pronounced stomatal and mesophyll limitations to CO2 transport, but quinoa retained photosynthetic capacity and photosystem II (PSII) performance. Saline water (300 mmol NaCl-equivalent to 60% of the salinity of sea-water) supplied in identical volumes to the irrigation received by the control and drought treatments induced similar reductions in stomatal and mesophyll conductance, but also reduced carboxylation of ribulose-1,5-bisphosphate carboxylase/oxygenase, regeneration of ribulose-1,5-bisphosphate, increased non-photochemical dissipation of energy as heat and impaired PSII electron transport. This suggests that ion toxicity reduced PN via interference with photosynthetic enzymes and degradation of pigment-protein complexes within the thylakoid membranes. The results of this study demonstrate that the photosynthetic physiology of quinoa is resistant to the effects of drought, but quinoa may not be a suitable crop for areas subject to strong salt stress or irrigation with a concentration of saline water equivalent to a 300 mmol NaCl solution.

Adaptation to high temperature mitigates the impact of water deficit during combined heat and drought stress in C3 sunflower and C4 maize varieties with contrasting drought tolerance.

Heat and drought stress frequently occur together, however, their impact on plant growth and photosynthesis (PN ) is unclear. The frequency, duration and severity of heat and drought stress events are predicted to increase in the future, having severe implications for agricultural productivity and food security. To assess the impact on plant gas exchange, physiology and morphology we grew drought tolerant and sensitive varieties of C3 sunflower (Helianthus annuus) and C4 maize (Zea mays) under conditions of elevated temperature for 4 weeks prior to the imposition of water deficit. The negative impact of temperature on PN was most apparent in sunflower. The drought tolerant sunflower retained ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) activity under heat stress to a greater extent than its drought sensitive counterpart. Maize exhibited no varietal difference in response to increased temperature. In contrast to previous studies, where a sudden rise in temperature induced an increase in stomatal conductance (Gs ), we observed no change or a reduction in Gs with elevated temperature, which alongside lower leaf area mitigated the impact of drought at the higher temperature. The drought tolerant sunflower and maize varieties exhibited greater investment in root-systems, allowing greater uptake of the available soil water. Elevated temperatures associated with heat-waves will have profound negative impacts on crop growth in both sunflower and maize, but the deleterious effect on PN was less apparent in the drought tolerant sunflower and both maize varieties. As C4 plants generally exhibit water use efficiency (WUE) and resistance to heat stress, selection on the basis of tolerance to heat and drought stress would be more beneficial to the yields of C3 crops cultivated in drought prone semi-arid regions.

Impaired Stomatal Control Is Associated with Reduced Photosynthetic Physiology in Crop Species Grown at Elevated [CO2].

Physiological control of stomatal conductance (Gs) permits plants to balance CO2-uptake for photosynthesis (PN) against water-loss, so optimizing water use efficiency (WUE). An increase in the atmospheric concentration of carbon dioxide ([CO2]) will result in a stimulation of PN and reduction of Gs in many plants, enhancing carbon gain while reducing water-loss. It has also been hypothesized that the increase in WUE associated with lower Gs at elevated [CO2] would reduce the negative impacts of drought on many crops. Despite the large number of CO2-enrichment studies to date, there is relatively little information regarding the effect of elevated [CO2] on stomatal control. Five crop species with active physiological stomatal behavior were grown at ambient (400 ppm) and elevated (2000 ppm) [CO2]. We investigated the relationship between stomatal function, stomatal size, and photosynthetic capacity in the five species, and then assessed the mechanistic effect of elevated [CO2] on photosynthetic physiology, stomatal sensitivity to [CO2] and the effectiveness of stomatal closure to darkness. We observed positive relationships between the speed of stomatal response and the maximum rates of PN and Gs sustained by the plants; indicative of close co-ordination of stomatal behavior and PN. In contrast to previous studies we did not observe a negative relationship between speed of stomatal response and stomatal size. The sensitivity of stomata to [CO2] declined with the ribulose-1,5-bisphosphate limited rate of PN at elevated [CO2]. The effectiveness of stomatal closure was also impaired at high [CO2]. Growth at elevated [CO2] did not affect the performance of photosystem II indicating that high [CO2] had not induced damage to the photosynthetic physiology, and suggesting that photosynthetic control of Gs is either directly impaired at high [CO2], sensing/signaling of environmental change is disrupted or elevated [CO2] causes some physical effect that constrains stomatal opening/closing. This study indicates that while elevated [CO2] may improve the WUE of crops under normal growth conditions, impaired stomatal control may increase the vulnerability of plants to water deficit and high temperatures.

Has the Impact of Rising CO2 on Plants been Exaggerated by Meta-Analysis of Free Air CO2 Enrichment Studies?

Meta-analysis is extensively used to synthesize the results of free air CO2 enrichment (FACE) studies to produce an average effect size, which is then used to model likely plant response to rising [CO2]. The efficacy of meta-analysis is reliant upon the use of data that characterizes the range of responses to a given factor. Previous meta-analyses of the effect of FACE on plants have not incorporated the potential impact of reporting bias in skewing data. By replicating the methodology of these meta-analytic studies, we demonstrate that meta-analysis of FACE has likely exaggerated the effect size of elevated [CO2] on plants by 20 to 40%; having significant implications for predictions of food security and vegetation response to climate change. Incorporation of the impact of reporting bias did not affect the significance or the direction of the [CO2] effect.

Coordination of stomatal physiological behavior and morphology with carbon dioxide determines stomatal control.

PREMISE OF THE STUDY: Stomatal control is determined by the ability to alter stomatal aperture and/or the number of stomata on the surface of new leaves in response to growth conditions. The development of stomatal control mechanisms to the concentration of CO2within the atmosphere ([CO2]) is fundamental to our understanding of plant evolutionary history and the prediction of gas exchange responses to future [CO2]. METHODS: In a controlled environment, fern and angiosperm species were grown in atmospheres of ambient (400 ppm) and elevated (2000 ppm) [CO2]. Physiological stomatal behavior was compared with the stomatal morphological response to [CO2]. KEY RESULTS: An increase in [CO2] or darkness induced physiological stomatal responses ranging from reductions (active) to no change (passive) in stomatal conductance. Those species with passive stomatal behavior exhibited pronounced reductions of stomatal density in new foliage when grown in elevated [CO2], whereas species with active stomata showed little morphological response to [CO2]. Analysis of the physiological and morphological stomatal responses of a wider range of species suggests that patterns of stomatal control to [CO2] do not follow a phylogenetic pattern associated with plant evolution. CONCLUSIONS: Selective pressures may have driven the development of divergent stomatal control strategies to increased [CO2]. Those species that are able to actively regulate guard cell turgor are more likely to respond to [CO2] through a change in stomatal aperture than stomatal number. We propose a model of stomatal control strategies in response to [CO2] characterized by a trade-off between short-term physiological behavior and longer-term morphological response.

Influence of olive oil solid waste applications on soil pH, electrical conductivity, soil nitrogen transformations, carbon content and aggregate stability.

Some of the agricultural wastes have important economic potentials to be used as a plant nutrition and/or soil conditioner. Olive oil solid waste (OSW) is one of them. The aim of this study was to determine effects of OSW on soil pH, electrical conductivity (EC), improving soil nitrogen (N) status, soil aggregate stability and soil carbon contents. Different rates of OSW (control, 2%, 4% and 8%, w/w) were applied to coarse soils and incubated for 2 months. OSW applications decreased soil pH values. The mean EC value of soil measured 2 months after the incubation was greater than that of the first month. Application of OSW at the rate of 8% has significantly increased soil total organic nitrogen contents. OSW applications at the rate of 8% (w/w) significantly increased both mean SOC content (3.5%) and aggregate stability (88%) after 2 months of incubation (p<0.01). Therefore, OSW has great potential to improve soil structure of coarse textured soil in short term.