Menachem Moshelion

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

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.

The Role of Tobacco Aquaporin1 in Improving Water Use Efficiency, Hydraulic Conductivity, and Yield Production Under Salt Stress

Tobacco (Nicotiana tabacum; C3) plants increase their water use efficiency (WUE) under abiotic stress and are suggested to show characteristics of C4 photosynthesis in stems, petioles, and transmitting tract cells. The tobacco stress-induced Aquaporin1 (NtAQP1) functions as both water and CO2 channel. In tobacco plants, overexpression of NtAQP1 increases leaf net photosynthesis (AN), mesophyll CO2 conductance, and stomatal conductance, whereas its silencing reduces root hydraulic conductivity (Lp). Nevertheless, interaction between NtAQP1 leaf and root activities and its impact on plant WUE and productivity under normal and stress conditions have never been suggested. Thus, the aim of this study was to suggest a role for NtAQP1 in plant WUE, stress resistance, and productivity. Expressing NtAQP1 in tomato (Solanum lycopersicum) plants (TOM-NtAQP1) resulted in higher stomatal conductance, whole-plant transpiration, and AN under all conditions tested. In contrast to controls, where, under salt stress, Lp decreased more than 3-fold, TOM-NtAQP1 plants, similar to maize (Zea mays; C4) plants, did not reduce Lp dramatically (only by approximately 40%). Reciprocal grafting provided novel evidence for NtAQP1s role in preventing hydraulic failure and maintaining the whole-plant transpiration rate. Our results revealed independent, albeit closely related, NtAQP1 activities in roots and leaves. This dual activity, which increases the plants water use and AN under optimal and stress conditions, resulted in improved WUE. Consequently, it contributed to the plants stress resistance in terms of yield production under all tested conditions, as demonstrated in both tomato and Arabidopsis (Arabidopsis thaliana) plants constitutively expressing NtAQP1. The putative involvement of NtAQP1 in tobaccos C4-like photosynthesis characteristics is discussed.

Diurnal and Circadian Regulation of Putative Potassium Channels in a Leaf Moving Organ

In a search for potassium channels involved in light- and clock-regulated leaf movements, we cloned four putative K channel genes from the leaf-moving organs, pulvini, of the legume Samanea saman. The S. saman SPOCK1 is homologous to KCO1, an Arabidopsis two-pore-domain K channel, the S. saman SPORK1 is similar to SKOR and GORK, Arabidopsis outward-rectifying Shaker-like K channels, and theS. saman SPICK1 and SPICK2 are homologous to AKT2, a weakly-inward-rectifying Shaker-like Arabidopsis K channel. All four S. saman sequences possess the universal K-channel-specific pore signature, TXXTXGYG, strongly suggesting a role in transmembrane K+ transport. The fourS. saman genes had different expression patterns within four leaf parts: ‘extensor’ and ‘flexor’ (the motor tissues), the leaf blades (mainly mesophyll), and the vascular bundle (‘rachis’). Based on northern blot analysis, their transcript level was correlated with the rhythmic leaf movements: (a) all four genes were regulated diurnally (Spick2, Spork1, andSpock1 in extensor and flexor, Spick1 in extensor and rachis); (b) Spork1 andSpock1 rhythms were inverted upon the inversion of the day-night cycle; and (c) in extensor and/or flexor, the expression ofSpork1, Spick1, and Spick2was also under a circadian control. These findings parallel the circadian rhythm shown to govern the resting membrane K+permeability in extensor and flexor protoplasts and the susceptibility of this permeability to light stimulation (Kim et al., 1993). Thus,Samanea pulvinar motor cells are the first described system combining light and circadian regulation of K channels at the level of transcript and membrane transport.

Hexokinase mediates stomatal closure

Stomata, composed of two guard cells, are the gates whose controlled movement allows the plant to balance the demand for CO2 for photosynthesis with the loss of water through transpiration. Increased guard-cell osmolarity leads to the opening of the stomata and decreased osmolarity causes the stomata to close. The role of sugars in the regulation of stomata is not yet clear. In this study, we examined the role of hexokinase (HXK), a sugar-phosphorylating enzyme involved in sugar-sensing, in guard cells and its effect on stomatal aperture. We show here that increased expression of HXK in guard cells accelerates stomatal closure. We further show that this closure is induced by sugar and is mediated by abscisic acid. These findings support the existence of a feedback-inhibition mechanism that is mediated by a product of photosynthesis, namely sucrose. When the rate of sucrose production exceeds the rate at which sucrose is loaded into the phloem, the surplus sucrose is carried toward the stomata by the transpiration stream and stimulates stomatal closure via HXK, thereby preventing the loss of precious water.

Role of aquaporins in determining transpiration and photosynthesis in water-stressed plants: crop water-use efficiency, growth and yield

The global shortage of fresh water is one of our most severe agricultural problems, leading to dry and saline lands that reduce plant growth and crop yield. Here we review recent work highlighting the molecular mechanisms allowing some plant species and genotypes to maintain productivity under water stress conditions, and suggest molecular modifications to equip plants for greater production in water-limited environments. Aquaporins (AQPs) are thought to be the main transporters of water, small and uncharged solutes, and CO2 through plant cell membranes, thus linking leaf CO2 uptake from the intercellular airspaces to the chloroplast with water loss pathways. AQPs appear to play a role in regulating dynamic changes of root, stem and leaf hydraulic conductivity, especially in response to environmental changes, opening the door to using AQP expression to regulate plant water-use efficiency. We highlight the role of vascular AQPs in regulating leaf hydraulic conductivity and raise questions regarding their role (as well as tonoplast AQPs) in determining the plant isohydric threshold, growth rate, fruit yield production and harvest index. The tissue- or cell-specific expression of AQPs is discussed as a tool to increase yield relative to control plants under both normal and water-stressed conditions.

Risk-taking plants: anisohydric behavior as a stress-resistance trait.

Water scarcity is a critical limitation for agricultural systems. Two different water management strategies have evolved in plants: an isohydric strategy and an anisohydric strategy. Isohydric plants maintain a constant midday leaf water potential (Ψleaf) when water is abundant, as well as under drought conditions, by reducing stomatal conductance as necessary to limit transpiration. Anisohydric plants have more variable Ψleaf and keep their stomata open and photosynthetic rates high for longer periods, even in the presence of decreasing leaf water potential. This risk-taking behavior of anisohydric plants might be beneficial when water is abundant, as well as under moderately stressful conditions. However, under conditions of intense drought, this behavior might endanger the plant. We will discuss the advantages and disadvantages of these two water-usage strategies and their effects on the plant’s ability to tolerate abiotic and biotic stress. The involvement of plant tonoplast AQPs in this process will also be discussed.

Dynamic changes in the osmotic water permeability of protoplast plasma membrane

The osmotic water permeability coefficient (Pf) of plasma membrane of maize (Zea mays) Black Mexican Sweet protoplasts changed dynamically during a hypoosmotic challenge, as revealed using a model-based computational approach. The best-fitting model had three free parameters: initial Pf, Pf rate-of-change (slopePf), and a delay, which were hypothesized to reflect changes in the number and/or activity of aquaporins in the plasma membrane. Remarkably, the swelling response was delayed 2 to 11 s after start of the noninstantaneous (but accounted for) bath flush. The Pf during the delay was ≤1 μm s−1. During the swelling period following the delay, Pf changed dynamically: within the first 15 s Pf either (1) increased gradually to approximately 8 μm s−1 (in the majority population of low-initial-Pf cells) or (2) increased abruptly to 10 to 20 μm s−1 and then decreased gradually to 3 to 6 μm s−1 (in the minority population of high-initial-Pf cells). We affirmed the validity of our computational approach by the ability to reproduce previously reported initial Pf values (including the absence of delay) in control experiments on Xenopus oocytes expressing the maize aquaporin ZmPIP2;5. Although mercury did not affect the Pf in swelling Black Mexican Sweet cells, phloretin, another aquaporin inhibitor, inhibited swelling in a predicted manner, prolonging the delay and slowing Pf increase, thereby confirming the hypothesis that Pf dynamics, delay included, reflected the varying activity of aquaporins.

Role of aquaporins in determining transpiration and photosynthesis in water-stressed plants

The global shortage of fresh water is one of our most severe agricultural problems, leading to dry and saline lands that reduce plant growth and crop yield. Here we review recent work highlighting the molecular mechanisms allowing some plant species and genotypes to maintain productivity under water stress conditions, and suggest molecular modifications to equip plants for greater production in water-limited environments. Aquaporins (AQPs) are thought to be the main transporters of water, small and uncharged solutes, and CO2 through plant cell membranes, thus linking leaf CO2 uptake from the intercellular airspaces to the chloroplast with water loss pathways. AQPs appear to play a role in regulating dynamic changes of root, stem and leaf hydraulic conductivity, especially in response to environmental changes, opening the door to using AQP expression to regulate plant water-use efficiency. We highlight the role of vascular AQPs in regulating leaf hydraulic conductivity and raise questions regarding their role (as well as tonoplast AQPs) in determining the plant isohydric threshold, growth rate, fruit yield production and harvest index. The tissue- or cell-specific expression of AQPs is discussed as a tool to increase yield relative to control plants under both normal and water-stressed conditions.

Differential tissue‑specific expression of NtAQP1 in Arabidopsis thaliana reveals a role for this protein in stomatal and mesophyll conductance of CO 2 under standard and salt‑stress conditions

The regulation of plant hydraulic conductance and gas conductance involves a number of different morphological, physiological and molecular mechanisms working in harmony. At the molecular level, aquaporins play a key role in the transport of water, as well as CO2, through cell membranes. Yet, their tissue-related function, which controls whole-plant gas exchange and water relations, is less understood. In this study, we examined the tissue-specific effects of the stress-induced tobacco Aquaporin1 (NtAQP1), which functions as both a water and CO2 channel, on whole-plant behavior. In tobacco and tomato plants, constitutive overexpression of NtAQP1 increased net photosynthesis (AN), mesophyll CO2 conductance (gm) and stomatal conductance (gs) and, under stress, increased root hydraulic conductivity (Lpr) as well. Our results revealed that NtAQP1 that is specifically expressed in the mesophyll tissue plays an important role in increasing both AN and gm. Moreover, targeting NtAQP1 expression to the cells of the vascular envelope significantly improved the plants’ stress response. Surprisingly, NtAQP1 expression in the guard cells did not have a significant effect under any of the tested conditions. The tissue-specific involvement of NtAQP1 in hydraulic and gas conductance via the interaction between the vasculature and the stomata is discussed.

Growth and physiological responses of isohydric and anisohydric poplars to drought

Highlight Isohydric poplars have high water-use efficiency, while anisohydric poplars show faster growth under a variable water supply, with implications for performance of the different genotypes for woody biomass production.