a-telemetric-gravimetric-platform-for-real-time-physiological

A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant&ndash;Environment Interactions

Food security for the growing global population is a major concern. The data provided by genomic tools far exceeds the supply of phenotypic data, creating ...

The Hebrew University of Jerusalem

The vertical diffusion of NaI solution from a horizontal fracture into and within the surrounding matrix was tracked and quantified over time using an artificially fractured chalk core (30x5 cm) and a second-generation X-ray computed tomography (CT) scanner. The different tracer-penetration distances imaged in the matrix above and below the horizontal fracture are indicative of a greater tracer mass penetrating into the lower matrix. The enhanced transport in the matrix below the fracture was related to the Rayleigh-Darcy instability induced by the density differences between the heavier tracer solution in the fracture (1.038) and the distilled water that had initially resided in the matrix. Our observations suggest that below the fracture, the tracer is propagated by an advection-diffusion process that is characterized by both higher rates and higher concentrations relative to its propagation by diffusion above the fracture. The experimental results suggest that the prediction of contaminant migration in a rock intersected by both vertical and horizontal (e.g. along bedding planes) fractures is difficult because of density effects that result in different solute-penetration rates.

Tracer diffusion from a horizontal fracture into the surrounding matrix: measurement by computed tomography.

Anisohydric plants are thought to be more drought tolerant than isohydric plants. However, the molecular mechanism determining whether the plant water potential during the day remains constant or not regardless of the evaporative demand (isohydric vs anisohydric plant) is not known. Here, it was hypothesized that aquaporins take part in this molecular mechanism determining the plant isohydric threshold. Using computational mining a key tonoplast aquaporin, tonoplast intrinsic protein 2;2 (SlTIP2;2), was selected within the large multifunctional gene family of tomato (Solanum lycopersicum) aquaporins based on its induction in response to abiotic stresses. SlTIP2;2-transformed plants (TOM-SlTIP2;2) were compared with controls in physiological assays at cellular and whole-plant levels. Constitutive expression of SlTIP2;2 increased the osmotic water permeability of the cell and whole-plant transpiration. Under drought, these plants transpired more and for longer periods than control plants, reaching a lower relative water content, a behavior characterizing anisohydric plants. In 3-yr consecutive commercial glasshouse trials, TOM-SlTIP2;2 showed significant increases in fruit yield, harvest index and plant mass relative to the control under both normal and water-stress conditions. In conclusion, it is proposed that the regulation mechanism controlling tonoplast water permeability might have a role in determining the whole-plant ishohydric threshold, and thus its abiotic stress tolerance.

Improving plant stress tolerance and yield production: is the tonoplast aquaporin SlTIP2;2 a key to isohydric to anisohydric conversion?

Plants respond to many environmental changes by rapidly adjusting their hydraulic conductivity and transpiration rate, thereby optimizing water-use efficiency and preventing damage due to low water potential. A multiple-load-cell apparatus, time-series analysis of the measured data, and residual low-pass filtering methods were used to monitor continuously and analyse transpiration of potted tomato plants (Solanum lycopersicum cv. Ailsa Craig) grown in a temperature-controlled greenhouse during well-irrigated and drought periods. A time derivative of the filtered residual time series yielded oscillatory behaviour of the whole plant's transpiration (WPT) rate. A subsequent cross-correlation analysis between the WPT oscillatory pattern and wet-wick evaporation rates (vertical cotton fabric, 0.14 m(2) partly submerged in water in a container placed on an adjacent load cell) revealed that autonomous oscillations in WPT rate develop under a continuous increase in water stress, whereas these oscillations correspond with the fluctuations in evaporation rate when water is fully available. The relative amplitude of these autonomous oscillations increased with water stress as transpiration rate decreased. These results support the recent finding that an increase in xylem tension triggers hydraulic signals that spread instantaneously via the plant vascular system and control leaf conductance. The regulatory role of synchronized oscillations in WPT rate in eliminating critical xylem tension points and preventing embolism is discussed.

Development of synchronized, autonomous, and self-regulated oscillations in transpiration rate of a whole tomato plant under water stress.

Understanding the role of geometry, inertia, and dynamic contact angle on wetting and dewetting of capillary tubes has theoretical and practical aspects alike. The specific and synergistic effects of these factors were studied theoretically using a mathematical model that includes inertial and dynamic contact angle terms. After validating the model for capillaries of uniform cross section, the model was extended to capillaries with sinusoidal modulations of the radius, since in practice, capillaries rarely have uniform cross-sections. The height of the meniscus during wetting and dewetting was significantly affected by the relations between the local slope of the capillary surface and the Young contact angle. Non-dimensional variables were defined using viscous effects and gravity as the scaling parameters. Simulations using the dimensionless model showed that the inertial and dynamic contact angle terms can be neglected for narrow capillaries of uniform cross-section but not for uniform, wide cross-section capillaries. Moreover, nonuniformity in cross-sectional area induced hysteresis, deceleration, blocking, and metastable equilibrium locations. An increase in contact angle further amplified the effect of geometry on wetting and dewetting processes. These results enable characterization and modeling of fluid retention and flow in porous structures that inherently consist of capillaries of varying cross section.

Synergistic effects of geometry, inertia, and dynamic contact angle on wetting and dewetting of capillaries of varying cross sections.

Asparagus bean (Vigna unguiculata ssp. sesquipedalis) is the Asian subspecies of cowpea, a drought-resistant legume crop native to Africa. In order to explore the genetic variation of drought responses in asparagus bean, we conducted multi-year phenotyping of drought resistance traits across the Chinese asparagus bean mini-core. The phenotypic distribution indicated that the ssp. sesquipedalis subgene pool has maintained high natural variation in drought responses despite known domestic bottleneck. Thirty-nine SNP loci were found to show an association with drought resistance via a genome-wide association study (GWAS). Whole-plant water relations were compared among four genotypes by lysimetric assay. Apparent genotypic differences in transpiration patterns and the critical soil water threshold in relation to dehydration avoidance were observed, indicating a delicate adaptive mechanism for each genotype to its own climate. Microarray gene expression analyses revealed that known drought resistance pathways such as the ABA and phosphate lipid signaling pathways are conserved between different genotypes, while differential regulation of certain aquaporin genes and hormonal genes may be important for the genotypic differences. Our results suggest that divergent sensitivity to soil water content is an important mechanism configuring the genotypic specific responses to water deficit. The SNP markers identified provide useful resources for marker-assisted breeding. 

Natural variation and gene regulatory basis for the responses of asparagus beans to soil drought.

We present a simple and effective high-throughput experimental platform for simultaneous and continuous monitoring of water relations in the soil-plant-atmosphere continuum of numerous plants under dynamic environmental conditions. This system provides a simultaneously measured, detailed physiological response profile for each plant in the array, over time periods ranging from a few minutes to the entire growing season, under normal, stress and recovery conditions and at any phenological stage. Three probes for each pot in the array and a specially designed algorithm enable detailed water-relations characterization of whole-plant transpiration, biomass gain, stomatal conductance and root flux. They also enable quantitative calculation of the whole plant water-use efficiency and relative water content at high resolution under dynamic soil and atmospheric conditions. The system has no moving parts and can fit into many growing environments. A screening of 65 introgression lines of a wild tomato species (Solanum pennellii) crossed with cultivated tomato (S. lycopersicum), using our system and conventional gas-exchange tools, confirmed the accuracy of the system as well as its diagnostic capabilities. The use of this high-throughput diagnostic screening method is discussed in light of the gaps in our understanding of the genetic regulation of whole-plant performance, particularly under abiotic stress.

High-throughput physiological phenotyping and screening system for the characterization of plant-environment interactions.

Regulation of the rate of transpiration is an important part of plants' adaptation to uncertain environments. Stomatal closure is the most common response to severe drought. By closing their stomata, plants reduce transpiration to better their odds of survival under dry conditions. Under mild to moderate drought conditions, there are several possible transpiration patterns that balance the risk of lost productivity with the risk of water loss. Here, we hypothesize that plant ecotypes that have evolved in environments characterized by unstable patterns of precipitation will display a wider range of patterns of transpiration regulation along with other quantitative physiological traits (QPTs), compared to ecotypes from less variable environments. We examined five accessions of wild barley (Hordeum vulgare ssp. spontaneum) from different locations in Israel (the B1K collection) with annual rainfall levels ranging from 100 to 900 mm, along with one domesticated line (cv. Morex). We measured several QPTs and morphological traits of these accessions under well-irrigated conditions, under drought stress and during recovery from drought. Our results revealed a correlation between precipitation-certainty conditions and QPT plasticity. Specifically, accessions from stable environments (very wet or very dry locations) were found to take greater risks in their water-balance regulation than accessions from areas in which rainfall is less predictable. Notably, less risk-taking genotypes recovered more quickly than more risk-taking ones once irrigation was resumed. We discuss the relationships between environment, polymorphism, physiological plasticity and fitness, and suggest a general risk-taking model in which transpiration-rate plasticity is negatively correlated with population polymorphism.

Risk-management strategies and transpiration rates of wild barley in uncertain environments.

The improvement of crop productivity under abiotic stress is one of the biggest challenges faced by the agricultural scientific community. Despite extensive research, the research-to-commercial transfer rate of abiotic stress-resistant crops remains very low. This is mainly due to the complexity of genotype × environment interactions and in particular, the ability to quantify the dynamic plant physiological response profile to a dynamic environment. Most existing phenotyping facilities collect information using robotics and automated image acquisition and analysis. However, their ability to directly measure the physiological properties of the whole plant is limited. We demonstrate a high-throughput functional phenotyping system (HFPS) that enables comparing plants' dynamic responses to different ambient conditions in dynamic environments due to its direct and simultaneous measurement of yield-related physiological traits of plants under several treatments. The system is designed as one-to-one (1:1) plant-[sensors+controller] units, i.e., each individual plant has its own personalized sensor, controller and irrigation valves that enable (i) monitoring water-relation kinetics of each plant-environment response throughout the plant's life cycle with high spatiotemporal resolution, (ii) a truly randomized experimental design due to multiple independent treatment scenarios for every plant, and (iii) reduction of artificial ambient perturbations due to the immobility of the plants or other objects. In addition, we propose two new resilience-quantifying-related traits that can also be phenotyped using the HFPS: transpiration recovery rate and night water reabsorption. We use the HFPS to screen the effects of two commercial biostimulants (a seaweed extract -ICL-SW, and a metabolite formula - ICL-NewFo1) on  under different irrigation regimes. Biostimulants are considered an alternative approach to improving crop productivity. However, their complex mode of action necessitates cost-effective pre-field phenotyping. The combination of two types of treatment (biostimulants and drought) enabled us to evaluate the precision and resolution of the system in investigating the effect of biostimulants on drought tolerance. We analyze and discuss plant behavior at different stages, and assess the penalty and trade-off between productivity and resilience. In this test case, we suggest a protocol for the screening of biostimulants' physiological mechanisms of action.

Article	Year
Tracer diffusion from a horizontal fracture into the surrounding matrix: measurement by computed tomography. Journal of contaminant hydrology\| PubMed ID: 14607472	2003
Improving plant stress tolerance and yield production: is the tonoplast aquaporin SlTIP2;2 a key to isohydric to anisohydric conversion? The New phytologist\| PubMed ID: 19054338	2009
Development of synchronized, autonomous, and self-regulated oscillations in transpiration rate of a whole tomato plant under water stress. Journal of experimental botany\| PubMed ID: 20558570	2010
Synergistic effects of geometry, inertia, and dynamic contact angle on wetting and dewetting of capillaries of varying cross sections. Journal of colloid and interface science\| PubMed ID: 23411356	2013
Natural variation and gene regulatory basis for the responses of asparagus beans to soil drought. Frontiers in plant science\| PubMed ID: 26579145	2015
High-throughput physiological phenotyping and screening system for the characterization of plant-environment interactions. The Plant journal : for cell and molecular biology\| PubMed ID: 27868265	2017
Risk-management strategies and transpiration rates of wild barley in uncertain environments. Physiologia plantarum\| PubMed ID: 30084486	2018
Dynamic Physiological Phenotyping of Drought-Stressed Pepper Plants Treated With "Productivity-Enhancing" and "Survivability-Enhancing" Biostimulants. Frontiers in plant science\| PubMed ID: 31379898	2019

Rony Wallach

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