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in Agriculture
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Publications

2021
Weksler, S. ; Rozenstein, O. ; Haish, N. ; Moshelion, M. ; Wallach, R. ; Ben-Dor, E. Detection of Potassium Deficiency and Momentary Transpiration Rate Estimation at Early Growth Stages Using Proximal Hyperspectral Imaging and Extreme Gradient Boosting. Sensors 2021, 21. Publisher's VersionAbstract
{Potassium is a macro element in plants that is typically supplied to crops in excess throughout the season to avoid a deficit leading to reduced crop yield. Transpiration rate is a momentary physiological attribute that is indicative of soil water content, the plant’s water requirements, and abiotic stress factors. In this study, two systems were combined to create a hyperspectral–physiological plant database for classification of potassium treatments (low, medium, and high) and estimation of momentary transpiration rate from hyperspectral images. PlantArray 3.0 was used to control fertigation, log ambient conditions, and calculate transpiration rates. In addition, a semi-automated platform carrying a hyperspectral camera was triggered every hour to capture images of a large array of pepper plants. The combined attributes and spectral information on an hourly basis were used to classify plants into their given potassium treatments (average accuracy = 80%) and to estimate transpiration rate (RMSE = 0.025 g/min
2020
Attia, Z. ; Dalal, A. ; Moshelion, M. Vascular bundle sheath and mesophyll cells modulate leaf water balance in response to chitin. The Plant JournalThe Plant JournalPlant J 2020, 101, 1368 - 1377. Publisher's VersionAbstract
Summary Plants can detect pathogen invasion by sensing microbe-associated molecular patterns (MAMPs). This sensing process leads to the induction of defense responses. Numerous MAMP mechanisms of action have been described in and outside the guard cells. Here, we describe the effects of chitin, a MAMP found in fungal cell walls and insects, on the cellular osmotic water permeability (Pf) of the leaf vascular bundle-sheath (BS) and mesophyll cells (MCs), and its subsequent effect on leaf hydraulic conductance (Kleaf). BS is a parenchymatic tissue that tightly encases the vascular system. BS cells (BSCs) have been shown to influence Kleaf through changes in their Pf, for example, after sensing the abiotic stress response-regulating hormone abscisic acid. It was recently reported that, in Arabidopsis, the chitin receptors-like kinases, chitin elicitor receptor kinase 1 (CERK1) and LYSINE MOTIF RECEPTOR KINASE 5 (LYK5) are highly expressed in the BS as well as the neighboring mesophyll. Therefore, we studied the possible impact of chitin on these cells. Our results revealed that BSCs and MCs exhibit a sharp decrease in Pf in response to chitin treatment. In addition, xylem-fed chitin decreased Kleaf and led to stomatal closure. However, Atlyk5 mutant showed none of these responses. Complementing AtLYK5 in the BSCs (using the SCARECROW promoter) resulted in the response to chitin that was similar to that observed in the wild-type. These results suggest that BS play a role in the perception of apoplastic chitin and in initiating chitin-triggered immunity.
Negin, B. ; Moshelion, M. Remember where you came from: ABA insensitivity is epigenetically inherited in mesophyll, but not seeds. Food Security under Climate Change 2020, 295, 110455. Publisher's VersionAbstract
Plants transmit their experiences of environmental conditions to their progeny through epigenetic inheritance, improving their progeny’s fitness under prevailing conditions. Though ABA is known to regulate epigenetic-modification genes, no strong phenotypic link between those genes and intergenerational “memory” has been shown. Previously, we demonstrated that mesophyll insensitivity to ABA (FBPase::abi1-1{fa} transgenic plants) results in a range of developmental phenotypes, including early growth vigor and early flowering (i.e., stress-escape behavior). Here, we show that null plants, used as controls (segregates of FBPase::abi1 that are homozygote descendants of a heterozygous transgenic plant, but do not contain the transformed abi1-1 gene) phenotypically resembled their FBPase::abi1-1 parents. However, in germination and early seedling development assays, null segregants resembled WT plants. These FBPase::abi1-1 null segregants mesophyll-related phenotypes were reproducible and stable for at least three generations. These results suggest that the heritability of stress response is linked to ABA’s epigenetic regulatory effect through ABI1 and mesophyll-related traits. The discrepancy between the epigenetic heritability of seed and mesophyll-related traits is an example of the complexity of epigenetic regulation, which is both gene and process-specific, and may be attributed to the fine-tuning of tradeoffs between flowering time, growth rate and levels of risk that allow annual plants to optimize their fitness in uncertain environments.
Moshelion, M. The dichotomy of yield and draught resistance. EMBO reports 2020, n/a, e51598. Publisher's VersionAbstract
Global climate change and the increasing human population require crop varieties with higher yield and draught resistance. But meeting both goals is not an easy task for breeders and plant science.
Dalal, A. ; Shenhar, I. ; Bourstein, R. ; Mayo, A. ; Grunwald, Y. ; Averbuch, N. ; Attia, Z. ; Wallach, R. ; Moshelion, M. A Telemetric, Gravimetric Platform for Real-Time Physiological Phenotyping of Plant–Environment Interactions. J. Vis. Exp. 2020, e61280. Publisher's VersionAbstract
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 a knowledge gap. To meet the challenge of improving crops to feed the growing global population, this gap must be bridged.Physiological traits are considered key functional traits in the context of responsiveness or sensitivity to environmental conditions. Many recently introduced high-throughput (HTP) phenotyping techniques are based on remote sensing or imaging and are capable of directly measuring morphological traits, but measure physiological parameters mainly indirectly. This paper describes a method for direct physiological phenotyping that has several advantages for the functional phenotyping of plant–environment interactions. It helps users overcome the many challenges encountered in the use of load-cell gravimetric systems and pot experiments. The suggested techniques will enable users to distinguish between soil weight, plant weight and soil water content, providing a method for the continuous and simultaneous measurement of dynamic soil, plant and atmosphere conditions, alongside the measurement of key physiological traits. This method allows researchers to closely mimic field stress scenarios while taking into consideration the environment’s effects on the plants’ physiology. This method also minimizes pot effects, which are one of the major problems in pre-field phenotyping. It includes a feed-back fertigation system that enables a truly randomized experimental design at a field-like plant density. This system detects the soil-water-content limiting threshold (θ) and allows for the translation of data into knowledge through the use of a real-time analytic tool and an online statistical resource. This method for the rapid and direct measurement of the physiological responses of multiple plants to a dynamic environment has great potential for use in screening for beneficial traits associated with responses to abiotic stress, in the context of pre-field breeding and crop improvement.
2019
Kelly, G. ; Egbaria, A. ; Khamaisi, B. ; Lugassi, N. ; Attia, Z. ; Moshelion, M. ; Granot, D. Guard-Cell Hexokinase Increases Water-Use Efficiency Under Normal and Drought Conditions. Frontiers in Plant Science 2019, 10. Publisher's VersionAbstract
Water is a limiting resource for many land plants. Most of the water taken up by plants is lost to the atmosphere through the stomata, which are adjustable pores on the leaf surface that allow for gas exchange between the plant and the atmosphere. Modulating stomatal activity might be an effective way to reduce plants’ water consumption and enhance their productivity under normal, as well as water-limiting conditions. Our recent discovery of stomatal regulation by sugars that is mediated by guard-cell hexokinase (HXK), a sugar-sensing enzyme, has raised the possibility that HXK might be used to increase plant water-use efficiency (WUE; i.e., carbon gain per unit of water). We show here that transgenic tomato and Arabidopsis plants with increased expression of HXK in their guard cells (GCHXK plants) exhibit reduced transpiration and higher WUE without any negative effects on growth under normal conditions, as well as drought avoidance and improved photosynthesis and growth under limited-water conditions. Our results demonstrate that exclusive expression of HXK in guard cells is an effective tool for improving WUE, and plant performance under drought. © Copyright © 2019 Kelly, Egbaria, Khamaisi, Lugassi, Attia, Moshelion and Granot.
Attia, Z. ; Dalal, A. ; Moshelion, M. Vascular bundle sheath and mesophyll cells modulate leaf water balance in response to chitin. Plant Journal 2019. Publisher's VersionAbstract
Plants can detect pathogen invasion by sensing microbe-associated molecular patterns (MAMPs). This sensing process leads to the induction of defense responses. Numerous MAMP mechanisms of action have been described in and outside the guard cells. Here, we describe the effects of chitin, a MAMP found in fungal cell walls and insects, on the cellular osmotic water permeability (Pf) of the leaf vascular bundle-sheath (BS) and mesophyll cells (MCs), and its subsequent effect on leaf hydraulic conductance (Kleaf). BS is a parenchymatic tissue that tightly encases the vascular system. BS cells (BSCs) have been shown to influence Kleaf through changes in their Pf, for example, after sensing the abiotic stress response-regulating hormone abscisic acid. It was recently reported that, in Arabidopsis, the chitin receptors-like kinases, chitin elicitor receptor kinase 1 (CERK1) and LYSINE MOTIF RECEPTOR KINASE 5 (LYK5) are highly expressed in the BS as well as the neighboring mesophyll. Therefore, we studied the possible impact of chitin on these cells. Our results revealed that BSCs and MCs exhibit a sharp decrease in Pf in response to chitin treatment. In addition, xylem-fed chitin decreased Kleaf and led to stomatal closure. However, Atlyk5 mutant showed none of these responses. Complementing AtLYK5 in the BSCs (using the SCARECROW promoter) resulted in the response to chitin that was similar to that observed in the wild-type. These results suggest that BS play a role in the perception of apoplastic chitin and in initiating chitin-triggered immunity. © 2019 The Authors The Plant Journal © 2019 John Wiley & Sons Ltd
Gosa, S. C. ; Lupo, Y. ; Moshelion, M. Quantitative and comparative analysis of whole-plant performance for functional physiological traits phenotyping: New tools to support pre-breeding and plant stress physiology studies. Plant Science 2019, 282, 49 - 59. Publisher's VersionAbstract
Plants are autotrophic organisms in which there are linear relationships between the rate at which organic biomass is accumulated and many ambient parameters such as water, nutrients, CO2 and solar radiation. These linear relationships are the result of good feedback regulation between a plants sensing of the environment and the optimization of its performance response. In this review, we suggest that continuous monitoring of the plant physiological profile in response to changing ambient conditions could be a useful new phenotyping tool, allowing the characterization and comparison of different levels of functional phenotypes and productivity. This functional physiological phenotyping (FPP) approach can be integrated into breeding programs, which are facing difficulties in selecting plants that perform well under abiotic stress. Moreover, high-throughput FPP will increase the efficiency of the selection of traits that are closely related to environmental interactions (such as plant water status, water-use efficiency, stomatal conductance, etc.) thanks to its high resolution and dynamic measurements. One of the important advantages of FPP is, its simplicity and effectiveness and compatibility with experimental methods that use load-cell lysimeters and ambient sensors. In the future, this platform could help with phenotyping of complex physiological traits, beneficial for yield gain to enhance functional breeding approaches and guide in crop modeling.
Dalal, A. ; Bourstein, R. ; Haish, N. ; Shenhar, I. ; Wallach, R. ; Moshelion, M. Dynamic Physiological Phenotyping of Drought-Stressed Pepper Plants Treated With "Productivity-Enhancing" and "Survivability-Enhancing" Biostimulants. Front Plant Sci 2019, 10, 905.Abstract
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.
Harfouche, A. L. ; Jacobson, D. A. ; Kainer, D. ; Romero, J. C. ; Harfouche, A. H. ; Scarascia Mugnozza, G. ; Moshelion, M. ; Tuskan, G. A. ; Keurentjes, J. J. B. ; Altman, A. Accelerating Climate Resilient Plant Breeding by Applying Next-Generation Artificial Intelligence. Trends Biotechnol 2019.Abstract
Breeding crops for high yield and superior adaptability to new and variable climates is imperative to ensure continued food security, biomass production, and ecosystem services. Advances in genomics and phenomics are delivering insights into the complex biological mechanisms that underlie plant functions in response to environmental perturbations. However, linking genotype to phenotype remains a huge challenge and is hampering the optimal application of high-throughput genomics and phenomics to advanced breeding. Critical to success is the need to assimilate large amounts of data into biologically meaningful interpretations. Here, we present the current state of genomics and field phenomics, explore emerging approaches and challenges for multiomics big data integration by means of next-generation (Next-Gen) artificial intelligence (AI), and propose a workable path to improvement.
Domec, J. - C. ; Berghoff, H. ; Way, D. ; Moshelion, M. ; Palmroth, S. ; Kets, K. ; Huang, C. - W. ; Oren, R. Mechanisms for minimizing height-related stomatal conductance declines in tall vines. Plant Cell Environ 2019.
Negin, B. ; Yaaran, A. ; Kelly, G. ; Zait, Y. ; Moshelion, M. Mesophyll ABA restrains early growth and flowering but does not directly suppress photosynthesis. Plant Physiol 2019.Abstract
Abscisic acid (ABA) levels increase significantly in plants under stress conditions and ABA is thought to serve as a key stress-response regulator. However, the direct effect of ABA on photosynthesis and the effect of mesophyll ABA on yield under both well-watered and drought conditions are still the subject of debate. Here, we examined this issue using transgenic Arabidopsis thaliana plants carrying a dominant ABA-signaling inhibitor under the control of a mesophyll-specific promoter (FBPase::abi1-1, abbreviated to fa). Under normal conditions, fa plants displayed slightly higher stomatal conductance and carbon assimilation than wild-type (WT) plants; however, these parameters were comparable following ABA treatment. These observations suggest that ABA does not directly inhibit photosynthesis in the short term. fa plants also exhibited a variety of altered phenotypes under optimal conditions, including more vigorous initial growth, earlier flowering, smaller flowers and delayed chlorophyll degradation. Furthermore, under optimal conditions, fa plant seed production was less than a third of that observed for the WT. However, under drought conditions, WT and fa seed yields were similar due to a significant reduction in WT seed and no reduction in fa seed. These findings suggest that endogenous basal ABA inhibits a stress-escape response under non-stress conditions, allowing plants to accumulate biomass and maximize yield. The lack of a correlation between flowering time and plant biomass combined with delayed chlorophyll degradation suggests that this stress-escape behavior is regulated independently and upstream of other ABA-induced effects such as rapid growth and flowering.
Yaaran, A. ; Negin, B. ; Moshelion, M. Role of guard-cell ABA in determining steady-state stomatal aperture and prompt vapor-pressure-deficit response. Plant Sci 2019, 281, 31-40.Abstract
Abscisic acid (ABA) is known to be involved in stomatal closure. However, its role in stomatal response to rapid increases in the vapor pressure deficit (VPD) is unclear. To study this issue, we generated guard cell-specific ABA-insensitive Arabidopsis plants (guard-cell specific abi1-1; GCabi). Under non-stressed conditions, the stomatal conductance (g) and apertures of GCabi plants were greater than those of control plants. This supports guard-cell ABA role as limiting steady-state stomatal aperture under non-stressful conditions. When there was a rapid increase in VPD (0.15 to 1 kPa), the g and stomatal apertures of GCabi decreased in a manner similar that observed in the WT control, but different from that observed in WT plants treated with fusicoccin. Low VPD increased the size of the stomatal apertures of the WT, but not of GCabi. We conclude that guard-cell ABA does not play a significant role in the initial, rapid stomatal closure that occurs in response to an increase in VPD, but is important for stomatal adaptation to ambient VPD. We propose a biphasic angiosperm VPD-sensing model that includes an initial ABA-independent phase and a subsequent ABA-dependent steady-state phase in which stomatal behavior is optimized for ambient VPD conditions.
2018
Fox, H. ; Doron-Faigenboim, A. ; Kelly, G. ; Bourstein, R. ; Attia, Z. ; Zhou, J. ; Moshe, Y. ; Moshelion, M. ; David-Schwartz, R. Transcriptome analysis of Pinus halepensis under drought stress and during recovery. Tree Physiology 2018, 38, 423-441. Publisher's VersionAbstract
Forest trees use various strategies to cope with drought stress and these strategies involve complex molecular mechanisms. Pinus halepensis Miller (Aleppo pine) is found throughout the Mediterranean basin and is one of the most drought-tolerant pine species. In order to decipher the molecular mechanisms that P. halepensis uses to withstand drought, we performed large-scale physiological and transcriptome analyses. We selected a mature tree from a semi-arid area with suboptimal growth conditions for clonal propagation through cuttings. We then used a high-throughput experimental system to continuously monitor whole-plant transpiration rates, stomatal conductance and the vapor pressure deficit. The transcriptomes of plants were examined at six physiological stages: pre-stomatal response, partial stomatal closure, minimum transpiration, post-irrigation, partial recovery and full recovery. At each stage, data from plants exposed to the drought treatment were compared with data collected from well-irrigated control plants. A drought-stressed P. halepensis transcriptome was created using paired-end RNA-seq. In total, ∼6000 differentially expressed, non-redundant transcripts were identified between drought-treated and control trees. Cluster analysis has revealed stress-induced down-regulation of transcripts related to photosynthesis, reactive oxygen species (ROS)-scavenging through the ascorbic acid (AsA)-glutathione cycle, fatty acid and cell wall biosynthesis, stomatal activity, and the biosynthesis of flavonoids and terpenoids. Up-regulated processes included chlorophyll degradation, ROS-scavenging through AsA-independent thiol-mediated pathways, abscisic acid response and accumulation of heat shock proteins, thaumatin and exordium. Recovery from drought induced strong transcription of retrotransposons, especially the retrovirus-related transposon Tnt1-94. The drought-related transcriptome illustrates this species' dynamic response to drought and recovery and unravels novel mechanisms. © The Author 2017.
Azoulay-Shemer, T. ; Schwankl, N. ; Rog, I. ; Moshelion, M. ; Schroeder, J. I. Starch biosynthesis by AGPase, but not starch degradation by BAM1/3 and SEX1, is rate-limiting for CO -regulated stomatal movements under short-day conditions. FEBS Lett 2018, 592, 2739-2759.Abstract
Starch in guard cells functions in osmoregulation during stomatal movements. Starch metabolism is controlled by the circadian clock. We investigated the role of starch metabolism in stomatal responses to CO under different photoperiodic conditions. Guard cell starch levels correlate with low/high [CO ] exposure. Starch biosynthesis-deficient AGPase (ADG1) mutants but, unexpectedly, not the starch degradation-deficient BAM1, BAM3, and SEX1 mutants alone, are rate-limiting for stomatal conductance responses to [CO ]-shifts. Interestingly, AGPase is rate-limiting solely under short- but not long-day conditions. These findings suggest a model of enhanced AGPase activity in guard cells under short days such that starch biosynthesis becomes rate-limiting for CO -induced stomatal closing.
Galkin, E. ; Dalal, A. ; Evenko, A. ; Fridman, E. ; Kan, I. ; Wallach, R. ; Moshelion, M. Risk-management strategies and transpiration rates of wild barley in uncertain environments. Physiol Plant 2018, 164, 412-428.Abstract
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.
2017
Negin, B. ; Moshelion, M. The advantages of functional phenotyping in pre-field screening for drought-tolerant crops. Functional Plant Biology 2017, 44, 107 - 118. Publisher's VersionAbstract
Increasing worldwide demand for food, feed and fuel presents a challenge in light of limited resources and climatic challenges. Breeding for stress tolerance and drought tolerance, in particular, is one the most challenging tasks facing breeders. The comparative screening of immense numbers of plant and gene candidates and their interactions with the environment represents a major bottleneck in this process. We suggest four key components to be considered in pre-field screens (phenotyping) for complex traits under drought conditions: (i) where, when and under which conditions to phenotype; (ii) which traits to phenotype; (iii) how to phenotype (which method); and (iv) how to translate collected data into knowledge that can be used to make practical decisions. We describe some common pitfalls, including inadequate phenotyping methods, incorrect terminology and the inappropriate use of non-relevant traits as markers for drought tolerance. We also suggest the use of more non-imaging, physiology-based, high-throughput phenotyping systems, which, used in combination with soil–plant–atmosphere continuum (SPAC) measurements and fitting models of plant responses to continuous and fluctuating environmental conditions, should be further investigated in order to serve as a phenotyping tool to better understand and characterise plant stress response. In the future, we assume that many of today’s phenotyping challenges will be solved by technology and automation, leaving us with the main challenge of translating large amounts of accumulated data into meaningful knowledge and decision making tools.
Aidoo, M. K. ; Quansah, L. ; Galkin, E. ; Batushansky, A. ; Wallach, R. ; Moshelion, M. ; Bonfil, D. J. ; Fait, A. A combination of stomata deregulation and a distinctive modulation of amino acid metabolism are associated with enhanced tolerance of wheat varieties to transient drought. Metabolomics 2017, 13, 138. Publisher's VersionAbstract
Mediterranean winter crops are commonly and increasingly exposed to irregular rainfall and high temperatures, which lead to transient drought events of different degrees, adversely affecting growth and yield. Hence, exploring the diverse degrees of tolerance to drought existing in the crop and the molecular strategies behind it is pivotal for the development of ad hoc breeding programs.
Fox, H. ; Doron-Faigenboim, A. ; Kelly, G. ; Bourstein, R. ; Attia, Z. ; Zhou, J. ; Moshe, Y. ; Moshelion, M. ; David-Schwartz, R. Transcriptome analysis of Pinus halepensis under drought stress and during recovery. Tree Physiology 2017, 38, 423-441. Publisher's VersionAbstract
Forest trees use various strategies to cope with drought stress and these strategies involve complex molecular mechanisms. Pinus halepensis Miller (Aleppo pine) is found throughout the Mediterranean basin and is one of the most drought-tolerant pine species. In order to decipher the molecular mechanisms that P. halepensis uses to withstand drought, we performed large-scale physiological and transcriptome analyses. We selected a mature tree from a semi-arid area with suboptimal growth conditions for clonal propagation through cuttings. We then used a high-throughput experimental system to continuously monitor whole-plant transpiration rates, stomatal conductance and the vapor pressure deficit. The transcriptomes of plants were examined at six physiological stages: pre-stomatal response, partial stomatal closure, minimum transpiration, post-irrigation, partial recovery and full recovery. At each stage, data from plants exposed to the drought treatment were compared with data collected from well-irrigated control plants. A drought-stressed P. halepensis transcriptome was created using paired-end RNA-seq. In total,  6000 differentially expressed, non-redundant transcripts were identified between drought-treated and control trees. Cluster analysis has revealed stress-induced down-regulation of transcripts related to photosynthesis, reactive oxygen species (ROS)-scavenging through the ascorbic acid (AsA)-glutathione cycle, fatty acid and cell wall biosynthesis, stomatal activity, and the biosynthesis of flavonoids and terpenoids. Up-regulated processes included chlorophyll degradation, ROS-scavenging through AsA-independent thiol-mediated pathways, abscisic acid response and accumulation of heat shock proteins, thaumatin and exordium. Recovery from drought induced strong transcription of retrotransposons, especially the retrovirus-related transposon Tnt1-94. The drought-related transcriptome illustrates this species’ dynamic response to drought and recovery and unravels novel mechanisms.
Kelly, G. ; Sade, N. ; Doron-Faigenboim, A. ; Lerner, S. ; Shatil-Cohen, A. ; Yeselson, Y. ; Egbaria, A. ; Kottapalli, J. ; Schaffer, A. A. ; Moshelion, M. ; et al. Sugar and hexokinase suppress expression of PIP aquaporins and reduce leaf hydraulics that preserves leaf water potential. Plant J 2017, 91, 325-339.Abstract
Sugars affect central aspects of plant physiology, including photosynthesis, stomatal behavior and the loss of water through the stomata. Yet, the potential effects of sugars on plant aquaporins (AQPs) and water conductance have not been examined. We used database and transcriptional analyses, as well as cellular and whole-plant functional techniques to examine the link between sugar-related genes and AQPs. Database analyses revealed a high level of correlation between the expression of AQPs and that of sugar-related genes, including the Arabidopsis hexokinases 1 (AtHXK1). Increased expression of AtHXK1, as well as the addition of its primary substrate, glucose (Glc), repressed the expression of 10 AQPs from the plasma membrane-intrinsic proteins (PIP) subfamily (PIP-AQPs) and induced the expression of two stress-related PIP-AQPs. The osmotic water permeability of mesophyll protoplasts of AtHXK1-expressing plants and the leaf hydraulic conductance of those plants were significantly reduced, in line with the decreased expression of PIP-AQPs. Conversely, hxk1 mutants demonstrated a higher level of hydraulic conductance, with increased water potential in their leaves. In addition, the presence of Glc reduced leaf water potential, as compared with an osmotic control, indicating that Glc reduces the movement of water from the xylem into the mesophyll. The production of sugars entails a significant loss of water and these results suggest that sugars and AtHXK1 affect the expression of AQP genes and reduce leaf water conductance, to coordinate sugar levels with the loss of water through transpiration.