Initial water deficit effects on Lupinus albus photosynthetic performance, carbon metabolism, and hormonal balance: metabolic reorganization prior to early stress responses

The early (2-4 d) effects of slowly imposed soil water deficit on Lupinus albus photosynthetic performance, carbon metabolism, and hormonal balance in different organs (leaf blade, stem stele, stem cortex, and root) were evaluated on 23-d-old plants (growth chamber assay). Our work shows that several metabolic adjustments occurred prior to alteration of the plant water status, implying that water deficit is perceived before the change in plant water status. The slow, progressive decline in soil water content started to be visible 3 d after withholding water (3 DAW). The earliest plant changes were associated with organ-specific metabolic responses (particularly in the leaves) and with leaf conductance and only later with plant water status and photosynthetic rate (4 DAW) or photosynthetic capacity (according to the Farquhar model; 6 DAW). Principal component analysis (PCA) of the physiological parameters, the carbohydrate and the hormone levels and their relative values, as well as leaf water-soluble metabolites full scan data (LC-MS/MS), showed separation of the different sampling dates. At 6 DAW classically described stress responses are observed, with plant water status, ABA level, and root hormonal balance contributing to the separation of these samples. Discrimination of earlier stress stages (3 and 4 DAW) is only achieved when the relative levels of indole-3-acetic acid (IAA), cytokinins (Cks), and carbon metabolism (glucose, sucrose, raffinose, and starch levels) are taken into account. Our working hypothesis is that, in addition to single responses (e.g. ABA increase), the combined alterations in hormone and carbohydrate levels play an important role in the stress response mechanism. Response to more advanced stress appears to be associated with a combination of cumulative changes, occurring in several plant organs. The carbohydrate and hormonal balance in the leaf (IAA to bioactive-Cks; soluble sugars to IAA and starch to IAA; relative abundances of the different soluble sugars) flag the initial responses to the slight decrease in soil water availability (10-15% decrease). Further alterations in sucrose to ABA and in raffinose to ABA relative values (in all organs) indicate that soil water availability continues to decrease. Such alterations when associated with changes in the root hormone balance indicate that the stress response is initiated. It is concluded that metabolic balance (e.g. IAA/bioactive Cks, carbohydrates/IAA, sucrose/ABA, raffinose/ABA, ABA/IAA) is relevant in triggering adjustment mechanisms.     

What determines the intracellular ATP concentration

Analysis is made of the mechanisms that control the intracellular ATP level. The balance between energy production and expenditure determines the energy charge of the cell and the ratio of [ATP] to the adenylate pool. The absolute ATP concentration is determined by the adenylate pool, which, in its turn, depends on the balance between the rates of AMP synthesis and degradation. Experimental data are discussed that demonstrate an increase in the adenylate pool in response to activation of energy-consuming processes. A hypothesis is proposed according to which variation in the adenylate pool and absolute ATP concentration affords a cell the possibility of additional control over processes fulfilling useful work. A mechanism involved in this regulation is described using human erythrocytes as an example. The hypothesis explains why different metabolic pathways (protein and DNA syntheses, polysaccharide synthesis, and lipid synthesis) use different trinucleotides (GTP, UTP, and CTP, respectively) as an energy source. This allows the cell to independently control these metabolic processes by varying the individual nucleotide pools.     

Saturation and Utilization of Nitrate Pools in Pea and Sugar Beet Leaves

The critical periods in the saturation of pea and sugar beet leaves with nitrate absorbed by roots were discriminated. In peas, during the first 14 h, all nitrate penetrating leaf cells was concentrated in the cytosol (metabolic pool). During the second period (14–62 h), nitrate began to flow into the vacuole (storage pool), and the filling of the metabolic pool continued. Metabolic pool was saturated by the end of this period (62 h). During the third period (62–110 h), further nitrate accumulation in the cell occurred because of expanding of the storage pool. Its saturation (similarly as total cell saturation) commenced 86 h after the start of nitrate uptake. In sugar beet leaves, both metabolic and storage nitrate pools were saturated by the end of the first period (14 h), and the sizes of these pools did not change during the second period (14–86 h). When pea plants were transferred to the nitrate-free medium, nitrate efflux began from the storage pool until its complete exhausting after 3 days. In sugar beet leaves, nitrate was still present in the storage pool 4 days after plant transfer to the nitrate-free medium. In both crops, nitrate export from the storage pool was aimed at the maintenance of the optimum nitrate concentration in the metabolic pool and, thus, at the maintenance of nitrate reductase activity. A functional diversity of nitrate compartmentation in the cells of various plant species is discussed.     

Modelling central metabolic fluxes by constraint‐based optimization reveals metabolic reprogramming of developing Solanum lycopersicum (tomato) fruit

Modelling of metabolic networks is a powerful tool to analyse the behaviour of developing plant organs, including fruits. Guided by our current understanding of heterotrophic metabolism of plant cells, a medium‐scale stoichiometric model, including the balance of co–factors and energy, was constructed in order to describe metabolic shifts that occur through the nine sequential stages of Solanum lycopersicum (tomato) fruit development. The measured concentrations of the main biomass components and the accumulated metabolites in the pericarp, determined at each stage, were fitted in order to calculate, by derivation, the corresponding external fluxes. They were used as constraints to solve the model by minimizing the internal fluxes. The distribution of the calculated fluxes of central metabolism were then analysed and compared with known metabolic behaviours. For instance, the partition of the main metabolic pathways (glycolysis, pentose phosphate pathway, etc.) was relevant throughout fruit development. We also predicted a valid import of carbon and nitrogen by the fruit, as well as a consistent CO₂ release. Interestingly, the energetic balance indicates that excess ATP is dissipated just before the onset of ripening, supporting the concept of the climacteric crisis. Finally, the apparent contradiction between calculated fluxes with low values compared with measured enzyme capacities suggest a complex reprogramming of the metabolic machinery during fruit development. With a powerful set of experimental data and an accurate definition of the metabolic system, this work provides important insight into the metabolic and physiological requirements of the developing tomato fruits.     

A chronobiological investigation of fitness for work in women on a 220 cal 24 h protein diet.

In a group of nine healthy but obese women a set of tests were used to estimate several physiological and psycho-physiological variables: self-rating of hunger, mood and physical vigor, self-recording of grip-strength, muscular fatigue test, eye-hand skill, tempo, etc. Measurements were performed every 6-hours, at fixed times, during 24-hours, once a week, before (non-restricted spontaneous food intake) and for three weeks on a 220 cal. special protein diet (calcium caseinate: 55g, associated with potassium salts, water and vitamins). Findings relating to the metabolic and hormonal variables while on this diet have already been reported: (a) The diet does not alter the circadian acrophase (peak) of the variables and (b) two distinct metabolic steps occur: first a peak of gluconeogenesis and second, a phase when ketone bodies and fatty acids are used almost exclusively as source of fuel. The present findings show that so long as nitrogen remains in balance this restricted diet (a) does not change the grip strength and the muscular fatigue test; (b) does not alter the mood, the physical vigor etc. and (c) induces a statistically significant fall in "hunger" (self-rating method). We conclude that this low protein diet is compatible with non-strenuous work.     

The challenges of cellular compartmentalization in plant metabolic engineering

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Highlights► Targeting enzymes to organelles with highest precursor pool affects product yield. ► Targeting of nonendogenous enzymes to organelles is a useful tool for engineering. ► Identification of transporters is central for effective metabolic engineering. ► Advances in metabolomics of isolated organelles will impact engineering strategies. ► Future plant metabolic engineering will include introduction of entire pathways.     

The interaction of cholesterol absorption and cholesterol synthesis in man

The total miscible pool of cholesterol in the body is determined largely by the interaction of cholesterol absorption and synthesis. In the present study we have examined the net effects of this interplay in one normal and five hypercholesteremic subjects when various amounts of cholesterol were made available for absorption. Feeding large amounts of cholesterol to the normocholesteremic patient caused an expansion of body pools by as much as 20 g before the amount of cholesterol re-excreted as fecal neutral steroids each day came into balance with the cholesterol absorbed from the diet. There was no detectable decrease in total body synthesis of cholesterol nor any increase in conversion of cholesterol into bile acids. However, feedback control of cholesterol synthesis was demonstrable when large quantities of plant sterols were fed: in the hypercholesteremic patients thus studied, the absorption of both endogenous and exogenous cholesterol was then greatly reduced, and a compensatory increase in synthesis occurred. Thus, the plant sterol experiments, but not the cholesterol feeding experiment, demonstrated that feedback control by dietary cholesterol does occur in man. That feedback control by dietary cholesterol is relatively unimportant in man seems to be due to the fact that in the metabolic "steady state" the absorption mechanism is essentially saturated by the large amounts of endogenous cholesterol available for reabsorption. These findings demonstrate that there are important differences between man and various laboratory animals in regard to the interaction of absorption and synthesis as factors controlling the size of tissue pools of cholesterol.     

Effect of acetate and octanoate on tricarboxylic acid cycle metabolite disposal during propionate oxidation in the perfused rat heart

Tricarboxylic acid cycle pool size is determined by anaplerosis and metabolite disposal. The regulation of the latter during propionate metabolism was studied in isolated perfused rat hearts in the light of the characteristics of NADP-linked malic enzyme, which is inhibited by acetyl-CoA. The acetyl-CoA concentration was varied by infusions of acetate and octanoate, and the rate of metabolite disposal was calculated from a metabolic balance sheet compiled from the relevant metabolic fluxes. Propionate addition increased the tricarboxylic acid cycle pool size 4-fold and co-infusion of acetate or octanoate did not change it further. Propionate caused a decrease in the CoA-SH concentration and a 10-fold increase in the propionyl-CoA concentration. A paradoxical increase in the CoA-SH concentration was observed upon co-infusion of acetate in the presence of propionate, an effect probably caused by competitive inhibition of propionate activation. A more pronounced decline in the propionyl-CoA concentration was observed upon the co-infusion of octanoate. In a metabolic steady state, acetate and octanoate reduced propionate disposal only slightly, but did not increase the tricarboxylic acid cycle pool size. The results are in accord with the notion that the tricarboxylic acid pool size is mainly regulated by the anaplerotic mechanisms.     

Oxygen isotope enrichment of organic matter in Ricinus communis during the diel course and as affected by assimilate transport

The oxygen isotope composition in leaf water and organic compounds in different plant tissues is useful for assessing the physiological performance of plants in their environment, but more information is needed on Delta(18)O variation during a diel course. Here, we assessed Delta(18)O of the organic matter in leaves, phloem and xylem in stem segments, and fine roots of Ricinus communis during a full diel cycle. Enrichment of newly assimilated organic matter in equilibrium with leaf water was calculated by applying a nonsteady-state evaporative enrichment model. During the light period, Delta(18)O of the water soluble organic matter pool in leaves and phloem could be explained by a 27 per thousand enrichment compared with leaf water enrichment. Leaf water enrichment influenced Delta(18)O of phloem organic matter during the night via daytime starch synthesis and night-time starch remobilization. Phloem transport did not affect Delta(18)O of phloem organic matter. Diel variation in Delta(18)O in organic matter pools can be modeled, and oxygen isotopic information is not biased during transport through the plant. These findings will aid field studies that characterize environmental influences on plant water balance using Delta(18)O in phloem organic matter or tree rings.     

Metabolic implications of stress-induced proline accumulation in plants

In many plants, free proline accumulates in response to the imposition of a wide range of biotic and abiotic stresses. Controversy has surrounded the extent to which this shift in nitrogen metabolism benefits plants under adverse environmental conditions. Most attempts to account for the phenomenon have focused on the ability of proline to mediate osmotic adjustment, stabilise subcellular structures and scavenge free radicals. However, often the cytoplasmic pool of free proline even after the imposition of stress is insufficient size to account for pronounced biophysical effects.Alternatively, selective preservation of this stress-induced response may relate to endpoints other than simply augmenting the cellular pool of free proline. Proline accumulation may reduce stress-induced cellular acidification or prime oxidative respiration to provide energy needed for recovery. High levels of proline synthesis during stress may maintain NAD(P)+/NAD(P)H ratios at values compatible with metabolism under normal conditions. Consideration of the cofactor preference of plant 1-pyrroline-5-carboxylate (P5C) reductase as well as the in vivo concentrations of the two pyridine nucleotide cofactors and their respective redox ratios suggests that even a small increase in proline biosynthesis might have a large impact on the level of reduction of the cellular NADP pool. The increased NADP+/NADPH ratio mediated by proline biosynthesis is likely to enhance activity of the oxidative pentose phosphate pathway. This would provide precursors to support the demand for increased secondary metabolite production during stress as well as nucleotide synthesis accompanying the accelerated rate of cell division upon relief from stress, when oxidation of proline is likely to provide an important energy source for ADP phosphorylation. Thus, the extreme sensitivity of the metabolic processes of proline synthesis and degradation themselves may be of benefit by regulating metabolic processes adversely affected by stress. This viewpoint is supported by consideration of other physiological phenomena not directly related to stress responses, but in which proline metabolism may also play a regulatory role.A mechanism is proposed whereby the interconversions of proline and P5C in different cell types and the associated transfer of redox potential between tissues may constitute a form of metabolic signalling within higher plants. Stress-related alterations in proline metabolism may impinge on systems of redox control of plant gene expression.     

Biosynthesis and Catabolism of L-Ascorbic Acid in Plants

L-Ascorbic acid (AsA) is a vital antioxidant compound that plays a critical role in the cellular metabolism of plants and animals. Research on plant AsA metabolism experienced a significant resurgence after 1998 following the identification of AsA-deficient Arabidopsis mutants and the elucidation of a biosynthetic pathway accepted by the overwhelming majority of the plant science community. The identification and cloning of novel biosynthetic genes and the ensuing metabolic engineering of plant AsA content has however revealed a more complex picture. Additional biosynthetic routes have been identified and unexpected biochemical phenotypes were observed upon expression of animal AsA biosynthetic genes. The isolation of novel AsA conjugates from plant tissues and the evidence for long distance transport of AsA in plants have provided additional facets to its functionality. Although some progress has been made regarding the impact of AsA recycling on pool size, we still do not have a clear picture of the biochemistry of AsA degradation. This communication comprehensively reviews new developments in the AsA metabolic system and prompts directions for future research.     

Hydraulic regulation strategies for whole-plant water balance of two maize inbred lines differing in drought resistance under short-term osmotic stress

The conservation of water in agriculture requires an understanding of the mechanisms of plant–water relations. This study aimed to reveal hydraulic regulation strategies of maize (Zea mays L.) for maintaining the plant water balance during drought. The water relations of two maize inbred lines (Tian4 and 478) that differ in their resistance to drought in the field were investigated under well-watered conditions and osmotic stress induced with 10 % PEG 6000. The leaf transpiration rate and leaf water potential of 478 varied diurnally, but remained constant in Tian4, which is more drought resistant. Tian4 plants showed morphological, anatomical and physiological advantages that protected them from foliar water loss. The strategies of leaf hydraulics to regulate leaf water balance during the day and during short-term osmotic stress also differed between Tian4 and 478. The leaf hydraulic conductivity of Tian4 and 478 increased temporarily, but their root hydraulic conductivities were reduced under osmotic stress. However, the root hydraulic conductivity of Tian4 subsequently recovered. Lower and rapidly reduced leaf transpiration and the ability of root hydraulics to recover from short-term osmotic stress can help explain the strategies for plant water balance of drought-tolerant maize.     

植物叶片水力与经济性状权衡关系的研究进展

叶片既是植物光合产物形成的主要场所, 又是整株植物的水力瓶颈、应对灾难性水力失调的安全阀门, 是植物碳水耦合权衡的重要器官。叶经济型谱反映了叶片经济性状“投资-收益”的权衡, 为验证植物进化过程中形成的物种对策提供了适用的理论框架。叶片水力性状变化会影响叶片经济性状及植物存活和生长。因此, 探索植物叶片水力与经济性状的权衡关系, 对建立植物碳-水耦合模型、揭示植物水-碳投资机理、扩展植物性状型谱等均有重要意义。该文首先综述了叶片水力性状、经济性状及两者之间的权衡关系, 分析了叶片导水率与水力脆弱性、失膨点水势、水容、安全阈值等水力性状以及与叶片的形态、结构和气体交换功能性状之间的关系。然后, 从叶片形态、解剖和叶脉网络结构以及气孔功能方面探讨了叶片水力性状与经济性状的调节机制。最后, 提出今后应加强三方面的研究: (1)探索建立植物根-茎-叶水力输导系统的碳-氮-水资源的整株经济型谱, 以揭示植物功能结构耦合、高效固碳用水的生理生态学机制; (2)探索叶片水力安全、水力效率和固碳效率之间的普适性权衡关系, 以深入理解抗旱植物叶片构建的生物物理结构与生理代谢的关系; (3)探索个体水平碳水代谢关系、水分运输与生长速率的耦合, 为代谢推演理论和植物群落尺度预测提供基础。     

Trade-offs between plant leaf hydraulic and economic traits

Leaf is the most important organ for carbon-water coupling of a plant because it is the primary medium for photosynthesis. It also acts as the hydraulic bottleneck and safety valve against hydraulic catastrophic dysfunctions. The leaf economics spectrum, which reflects the balance between investments and returns of leaf economic traits, provides a useful framework for examining species strategies as shaped by their evolutionary history. Changes in leaf hydraulic traits will influence leaf economic traits as well as plant survival and growth. Exploring trade-offs between leaf hydraulic and economic traits is thus of significance for modeling carbon-water relations, understanding the mechanisms of water/carbon investments, and extending the leaf economic spectrum. In this review, we first examined the trade-offs between leaf hydraulic and economic traits. Specially, we analyzed the relationships between leaf hydraulic conductivity and hydraulic vulnerability, water potential at the turgor loss point, water capacitance, safety margin, and leaf morphological, structural and functional traits. We then discussed potential mechanisms regulating leaf hydraulic and economic traits from leaf morphology, anatomy, venation, and stomatal functions. Finally, we proposed future research to: (1) develop an integrated whole-plant economics spectrum, including carbon-nitrogen-water resources and root-stem-leaf hydraulic transport system that will help revealing ecophysiological mechanisms of plant structure-functional coupling, carbon sequestration and water use; (2) explore a generalized trade-offs among leaf hydraulic safety, hydraulic efficiency and carbon fixation efficiency to advance our understanding of the relationships between biophysical structure and physiological metabolism in plant leaf construction under drought stress; and (3) explore the carbon-water metabolic relationship and coupling of water transport and growth rate for the metabolic theory and predictions at community scale.     

Genome-wide identification and characterization of aquaporin gene family in moso bamboo (<Emphasis Type="Italic">Phyllostachys edulis</Emphasis>)

Aquaporins (AQPs) are known to play a major role in maintaining water and hydraulic conductivity balance in the plant system. Numerous studies have showed AQPs execute multi-function throughout plant growth and development, including water transport, nitrogen, carbon, and micronutrient acquisition etc. However, little information on AQPs is known in bamboo. In this study, we present the first genome-wide identification and characterization of AQP genes in moso bamboo (Phyllostachys edulis) using bioinformatics. In total, 26 AQP genes were identified by homologous analysis, which were divided into four groups (PIPs, TIPs, NIPs, and SIPs) based on the phylogenetic analysis. All the genes were located on 26 different scaffolds respectively on basis of the gene mapped to bamboo genome. Evolutionary analysis indicated that Ph. edulis was more close to Oryza sativa than Zea mays in the genetic relationship. Besides, qRT-PCR was used to analyze gene expression profiles, which revealed that AQP genes were expressed constitutively in all the detected tissues, and were all responsive to the environmental cues such as drought, water, and NaCl stresses. This data suggested that AQPs may play fundamental roles in maintaining normal growth and development of bamboo, which would contribute to better understanding for the complex regulation mechanism involved in the fast-growing process of bamboo. Furthermore, the result could provide valuable information for further research on bamboo functional genomics.     

Mutation-Selection Balance and Metabolic Control Theory

The evolution of metabolic control is examined with models that unify approaches of classical quantitative genetics and metabolic control theory. The quantitative traits considered are the activities of enzymes embedded within metabolic pathways. In the models, polygenic mutation alters the enzyme activities (Vmax/Km) according to prescribed distributions, and the population evolves following classical haploid viability selection. Stabilizing selection operates on global properties of the metabolic pathway, including either flux or metabolite pool concentration. Analytical results and numerical simulations demonstrate several important properties of these characters, including skewed, non-Gaussian equilibrium distributions, and an expected positive correlation between activities of enzymes flanking a substrate pool undergoing stabilizing selection. The house-of-cards approximation proved to be accurate in predicting the equilibrium distribution of allelic effects for a biologically reasonable segment of the parameter space. Further experimental and theoretical work is needed before a clear assessment can be made whether the observed variance in enzyme activities is explicable by a mutation-selection balance, and this system provides an excellent opportunity for such a test.     

Experimental cloud immersion and foliar water uptake in saplings of Abies fraseri and Picea rubens

Key message

Frequent cloud immersion events result in direct uptake of cloud water and improve plant water potentials during daylight hours in saplings of two dominant cloud forest species.

Abstract

In ecosystems with frequent cloud immersion, the influence on plant water balance can be important. While cloud immersion can reduce plant water loss via transpiration, recent advances in methodology have suggested that many species also absorb water directly into leaves (foliar water uptake). The current study examines foliar water uptake and its influence on daily plant water balance in tree species of the endangered spruce–fir forest of the southern Appalachian Mountains, USA. These mountain-top communities are considered relic, boreal forests that may have persisted because of the benefits of frequent cloud immersion. We examined changes in needle water content, xylem water potentials, and stable isotope values in saplings of the two dominant tree species, Abies fraseri and Picea rubens before and after a 24 h period of experimental cloud immersion. Both species exhibited foliar water uptake following immersion, evidenced by substantial changes in stable isotope values of extracted needle water that reflected the composition of the fog water. In addition, total needle water content improved 3.7–6.4 % following experimental submersion and xylem water potentials were significantly greater (up to 0.33 MPa) in cloud-immersed plants over control plants. These results indicate that foliar water uptake may be an adaptive strategy for utilizing cloud water and improving overall tree vigor in these most southerly distributed boreal species.     

The input of the chemical potential gradient of the water in the medium to the energy balance of terrestrial vascular plants

One of the characteristic feature of activity of plant organism vital functions is a speed of its inner water flow. The main source of energy providing the transport of water is gradient of water chemical potential between soil and atmosphere. In case of equal potentials of these surroundings, the plant organism have additional energetic expenses to compensate the energy of external source. Series of experiments to investigate the influence of saturation of atmosphere with water (equilibrium of potentials) to energetic balance of plant were carried out. The results shows that: 1) etiolate seedlings in this conditions waste 10-15 percent energy per unit of new formation more than under 70-80 percent humidity; 2) saturation of atmosphere with water above plants with well developed transpiration surface leads to the decrease in their heat content; 3) equilibrium of water chemical potentials of different environments increases the intensity of intact root transpiration, i.e. intensifies "the burning" of organic matter. Thus, gradient of water chemical potential existing in nature between environments of different organs if sufficient source of energy for a plant.     

Photostasis and cold acclimation: sensing low temperature through photosynthesis

Photosynthesis is a highly integrated and regulated process which is highly sensitive to any change in environmental conditions, because it needs to balance the light energy absorbed by the photosystems with the energy consumed by metabolic sinks of the plant. Low temperatures exacerbate an imbalance between the source of energy and the metabolic sink, thus requiring adjustments of photosynthesis to maintain the balance of energy flow. Photosynthesis itself functions as a sensor of this imbalance through the redox state of photosynthetic electron-transport components and regulates photophysical, photochemical and metabolic processes in the chloroplast. Recent progress has been made in understanding how plants sense the low temperature signal. It is clear that photosynthesis interacts with other processes during cold acclimation involving crosstalk between photosynthetic redox, cold acclimation and sugar-signalling pathways to regulate plant acclimation to low temperatures.