Summary
Experimental studies on CO2 assimilation of mesophytic C3 plants in relation to relative water content (RWC) are discussed. Decreasing RWC slows the actual rate of photosynthetic CO2 assimilation (A) and decreases the potential rate (Apot). Generally, as RWC falls from c. 100 to c. 75%, the stomatal conductance (gs) decreases, and with it A. However, there are two general types of relation of Apot to RWC, which are called Type 1 and Type 2. Type 1 has two main phases. As RWC decreases from 100 to c. 75%, Apot is unaffected, but decreasing stomatal conductance (gs) results in smaller A, and lower CO2 concentration inside the leaf (Ci) and in the chloroplast (Cc), the latter falling possibly to the compensation point. Down-regulation of electron transport occurs by energy quenching mechanisms, and changes in carbohydrate and nitrogen metabolism are considered acclimatory, caused by low Ci and reversible by elevated CO2. Below 75% RWC, there is metabolic inhibition of Apot, inhibition of A then being partly (but progressively less) reversible by elevated CO2; gs regulates A progressively less, and Ci and CO2 compensation point, Γ rise. It is suggested that this is the true stress phase, where the decrease in Apot is caused by decreased ATP synthesis and a consequent decreased synthesis of RuBP. In the Type 2 response, Apot decreases progressively at RWC 100 to 75%, with A being progressively less restored to the unstressed value by elevated CO2. Decreased gs leads to a lower Ci and Cc but they probably do not reach compensation point: gs becomes progressively less important and metabolic limitations more important as RWC falls. The primary effect of low RWC on Apot is most probably caused by limited RuBP synthesis, as a result of decreased ATP synthesis, either through inhibition of Coupling Factor activity or amount due to increased ion concentration. Carbohydrate synthesis and accumulation decrease. Type 2 response is considered equivalent to Type 1 at RWC below c. 75%, with Apot inhibited by limited ATP and RuBP synthesis, respiratory metabolism dominates and Ci and Γ rise. The importance of inhibited ATP synthesis as a primary cause of decreasing Apot is discussed. Factors determining the Type 1 and Type 2 responses are unknown. Electron transport is maintained (but down-regulated) in Types 1 and 2 over a wide range of RWC, and a large reduced/oxidized adenylate ratio results. Metabolic imbalance results in amino acid accumulation and decreased and altered protein synthesis. These conditions profoundly affect cell functions and ultimately cause cell death. Type 1 and 2 responses may reflect differences in gs and in sensitivity of metabolism to decreasing RWC.

https://onlinelibrary.wiley.com/doi/pdf/10.1046/j.0016-8025.2001.00814.x

Photosynthetic carbon assimilation and associated metabolism in relation to water deficits in higher plants

Plants regularly face adverse growth conditions, such as drought, salinity, chilling, freezing, and high temperatures. These stresses can delay growth and development, reduce productivity, and, in extreme cases, cause plant death. Plant stress responses are dynamic and involve complex cross-talk between different regulatory levels, including adjustment of metabolism and gene expression for physiological and morphological adaptation. In this review, information about metabolic regulation in response to drought, extreme temperature, and salinity stress is summarized and the signalling events involved in mediating stress-induced metabolic changes are presented.

Drought, salt, and temperature stress-induced metabolic rearrangements and regulatory networks

Dramatic accumulation of proline due to increased synthesis and decreased degradation under a variety of stress conditions such as salt, drought and metal has been documented in many plants. Similarly, a decrease in the level of accumulated proline in the rehydrated plants is due to both down regulation of proline biosynthetic pathway enzymes and upregulation of proline degrading enzymes. But, the role of proline during plant development and the molecular basis of the effect of proline accumulation during stress and upon relief of stress are still largely obscure. Here, we summarize the genes governing the proline biosynthetic pathway, its degradation and regulation. Sequentially, we provide an account on transgenics raised so far to engineer the overproduction of osmolyte proline. Also, the identification of specific cellular pathways involved in proline biosynthesis and metabolic changes occurring in transgenic plants developed for proline enhancements are discussed. Further, emphasis is also made on an untouched area of signal transduction of proline biosynthetic pathway.

Regulation of proline biosynthesis, degradation, uptake and transport in higher plants: Its implications in plant growth and abiotic stress tolerance

Polyamines and environmental challenges : recent development

Plant metabolism is perturbed by various abiotic stresses. As such the metabolic network of plants must be reconfigured under stress conditions in order to allow both the maintenance of metabolic homeostasis and the production of compounds that ameliorate the stress. The recent development and adoption of metabolomics and systems biology approaches enable us not only to gain a comprehensive overview, but also a detailed analysis of crucial components of the plant metabolic response to abiotic stresses. In this review we introduce the analytical methods used for plant metabolomics and describe their use in studies related to the metabolic response to water, temperature, light, nutrient limitation, ion and oxidative stresses. Both similarity and specificity of the metabolic responses against diverse abiotic stress are evaluated using data available in the literature. Classically discussed stress compounds such as proline, γ-amino butyrate and polyamines are reviewed, and the widespread importance of branched chain amino acid metabolism under stress condition is discussed. Finally, where possible, mechanistic insights into metabolic regulatory processes are discussed.

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The use of metabolomics to dissect plant responses to abiotic stresses

Thellungiella salsuginea, a Brassicaceae species closely related to Arabidopsis thaliana, is tolerant to high salinity. The two species were compared under conditions of osmotic stress to assess the relationships between stress tolerance, the metabolome, water homeostasis and growth performance. A broad range of metabolites were analysed by metabolic fingerprinting and profiling, and the results showed that, despite a few notable differences in raffinose and secondary metabolites, the same metabolic pathways were regulated by salt stress in both species. The main difference was quantitative: Thellungiella had much higher levels of most metabolites than Arabidopsis whatever the treatment. Comprehensive quantification of organic and mineral solutes showed a relative stability of the total solute content regardless of the species or treatment, meaning that little or no osmotic adjustment occurred under stress. The reduction in osmotic potential observed in plants under stress was found to result from a passive loss of water. Thellungiella shoots contain less water than Arabidopsis shoots, and have the ability to lose more water, which could contribute to maintain a water potential gradient between soil and plant. Significant differences between Thellungiella and Arabidopsis were also observed in terms of the physicochemical properties of their metabolomes, such as water solubility and polarity. On the whole, the Thellungiella metabolome appears to be more compatible with dehydration. Osmotic stress was also found to impact the metabolome properties in both species, increasing the overall polarity. Together, the results suggest that Thellungiella copes with osmotic stress by tolerating dehydration, with its metabolic configuration lending itself to osmoprotective strategies rather than osmo-adjustment.

https://www.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-313X.2010.04323.x

Metabolome and water homeostasis analysis of Thellungiella salsuginea suggests that dehydration tolerance is a key response to osmotic stress in this halophyte

Large amounts of nitrogen (N) fertilizers are used in the production of oilseed rape. However, as low-input methods of crop management are introduced crops will need to withstand temporary N deficiency. In temperate areas, oilseed rape will also be affected by frequent drought periods. Here we evaluated the physiological and metabolic impact of nitrate limitation on the oilseed rape response to water deprivation. Different amounts of N fertilizer were applied to plants at the vegetative stage, which were then deprived of water and rehydrated. Both water and N depletion accelerated leaf senescence and reduced leaf development. N-deprived plants exhibited less pronounced symptoms of wilting during drought, probably because leaves were smaller and stomata were partially closed. Efficiency of proline production, a major stress-induced diversion of nitrogen metabolism, was assessed at different positions along the whole plant axis and related to leaf developmental stage and water status indices. Proline accumulation, preferentially in younger leaves, accounted for 25–85 % of the free amino acid pool. This was mainly due to a better capacity for proline synthesis in fully N-supplied plants whether they were subjected to drought or not, as deduced from the expression patterns of the proline metabolism BnP5CS and BnPDH genes. Although less proline accumulated in the oldest leaves, a significant amount was transported from senescing to emerging leaves. Moreover, during rehydration proline was readily recycled. Our results therefore suggest that proline plays a significant role in leaf N remobilization and in N use efficiency in oilseed rape.

https://link.springer.com/content/pdf/10.1007%2Fs00425-012-1636-8.pdf

Nitrogen availability impacts oilseed rape (Brassica napus L.) plant water status and proline production efficiency under water-limited conditions

The fate of osmo-accumulated proline in leaf discs of Rape (Brassica napus L.) incubated in a medium of low osmolarity

Metabolomic investigation of the freezing-tolerant Arabidopsis mutant esk1 revealed large alterations in polar metabolite content in roots and shoots. Stress metabolic markers were found to be among the most significant metabolic markers associated with the mutation, but also compounds related to growth regulation or nutrition. The metabolic phenotype of esk1 was also compared to that of wild type (WT) under various environmental constraints, namely cold, salinity and dehydration. The mutant was shown to express constitutively a subset of metabolic responses which fits with the core of stress metabolic responses in the WT. But remarkably, the most specific metabolic responses to cold acclimation were not phenocopied by esk1 mutation and remained fully inducible in the mutant at low temperature. Under salt stress, esk1 accumulated lower amounts of Na(+) in leaves than the WT, and under dehydration stress its metabolic profile and osmotic potential were only slightly impacted. These phenotypes are consistent with the hypothesis of an altered water status in esk1, which actually exhibited basic lower water content (WC) and transpiration rate (TR) than the WT. Taken together, the results suggest that ESK1 does not function as a specific cold acclimation gene, but could rather be involved in water homeostasis.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-3040.2008.01898.x

Metabolome and water status phenotyping of Arabidopsis under abiotic stress cues reveals new insight into ESK1 function.

The relationships between the changes induced in abscisic acid (ABA) and proline contents were investigated in canola (Brassica napus L.) leaf discs (CLDs) subjected to sequential hyper-and hypo-osmotic treatments. Changes in ABA content were found to precede that of proline, suggesting that ABA could exert regulatory effects on both osmo-induced proline accumulation and its subsequent mobilization during recovery. When exogenously supplied in the light during recovery, ABA caused inhibition of the apparent rate of proline mobilization with an I 50 of about 20 μM. Under dark conditions, the ABA effect was not observed. This is in accordance with the fact that this effect could be suppressed in the light while using photosynthesis inhibitors. The ABA effect was partly restored under dark conditions when the CLDs were supplied with NADPH at high concentration. Under both conditions, ABA exerted its effect when supplied together with cycloheximide, suggesting that it does not rely on protein synthesis. When turgid CLDs, which heavily absorbed exogenously supplied L-proline, were incubated in the light on the reference medium with ABA, proline mobilization was also restricted, provided ABA was supplied during the period of proline uptake. Surprisingly, in CLDs that had recovered their full turgor after application of successive hyper- and hypo-osmotic treatments, application of ABA led to a very high accumulation of proline. In contrast, in freshly cut turgid CLDs incubated in presence of ABA, the basic proline level was only weakly enhanced. Thus, the apparent inhibitory effect of ABA on proline mobilization could be mediated through both an activation of biosynthesis and an inhibition of catabolism of this amino acid via light-dependent processes that remain to be elucidated.

Marie-Françoise Niogret

Papers

Metabolome and water homeostasis analysis of Thellungiella salsuginea suggests that dehydration tolerance is a key response to osmotic stress in this halophyte

Nitrogen availability impacts oilseed rape (Brassica napus L.) plant water status and proline production efficiency under water-limited conditions

The fate of osmo-accumulated proline in leaf discs of Rape (Brassica napus L.) incubated in a medium of low osmolarity

Metabolome and water status phenotyping of Arabidopsis under abiotic stress cues reveals new insight into ESK1 function.

Proline level is partly under the control of abscisic acid in canola leaf discs during recovery from hyper‐osmotic stress