The calcium-dependent protein kinase CPK7 acts on root hydraulic conductivity.

The effects of abscisic acid (ABA) on aquaporin content, root hydraulic conductivity (Lpr), whole plant hydraulic conductance, and leaf growth are controversial. We addressed these effects via a combination of experiments at different scales of plant organization and tested their consistency via a model. We analyzed under moderate water deficit a series of transformed maize (Zea mays) lines, one sense and three antisense, affected in NCED (for 9-cis-epoxycarotenoid dioxygenase) gene expression and that differed in the concentration of ABA in the xylem sap. In roots, the mRNA expression of most aquaporin PIP (for plasma membrane intrinsic protein) genes was increased in sense plants and decreased in antisense plants. The same pattern was observed for the protein contents of four PIPs. This resulted in more than 6-fold differences between lines in Lpr under both hydrostatic and osmotic gradients of water potential. This effect was probably due to differences in aquaporin activity, because it was nearly abolished by a hydrogen peroxide treatment, which blocks the water channel activity of aquaporins. The hydraulic conductance of intact whole plants was affected in the same way when measured either in steady-state conditions or via the rate of recovery of leaf water potential after rewatering. The recoveries of leaf water potential and elongation upon rehydration differed between lines and were accounted for by the experimentally measured Lpr in a model of water transfer. Overall, these results suggest that ABA has long-lasting effects on plant hydraulic properties via aquaporin activity, which contributes to the maintenance of a favorable plant water status.

Plant water transport and aquaporins in oxygen-deprived environments

Plant hydraulic conductivity (K) refers to the rate of water flow (kg s−1) per unit pressure drop (MPa), which drives flow through the plant organ system. It is an important eco-physiology index for measuring plant water absorption and transport capacity. A field study was conducted in the arid region of the Heihe River Basin in northwestern China, plant hydraulic conductivity was measured by high-pressure flowmeter (HPFM) to investigate the characteristics of hydraulic conductivity of typical dominant desert plants (Reaumuria soongarica M., Nitraria sphaerocarpa M., and Sympegma regelii B.) and their relationship with functional traits of leaves, stems, and roots, and explaining their adaptation strategies to desert environment from the perspective of plant organs hydraulic conductivity. The results showed that the hydraulic conductivity of the leaves and stems of R. soongarica and N. sphaerocarpa (KLA, leaf hydraulic conductivity per unit leaf area; KLW, leaf hydraulic conductivity per unit leaf weight; KSLA, stem hydraulic conductivity per unit leaf area; KSLW, stem hydraulic conductivity per unit leaf weight) were significantly lower than those of S. regelii, while their fine root (KRL, root hydraulic conductivity per unit leaf length; KRSA, root hydraulic conductivity per unit root surface area) and whole root (KTRW, whole root hydraulic conductivity per unit root weight) of hydraulic conductivity were significantly higher than those of S. regelii. In addition, KLA and KLW, KSLA and KSLW, and KRL and KRSA in three desert plants all exhibited consistent trends. Correlation analysis illustrated that the hydraulic conductivity of leaves and stems had a significantly positive correlation, but they had no significant negative correlation with the specific leaf weight (SLW, specific leaf weight). The hydraulic conductivity of fine root weight (KRW, root hydraulic conductivity per unit root weight) and specific root surface area (SRSA, specific root surface area) showed significantly positive correlation (r = 0.727, P < 0.05). The results demonstrated that the R. soongarica and N. sphaerocarpa preserved their water content through the strong leaf absorption capacity of soil water and the low water dispersion rates of leaves to adapt to the harsher arid habitat, which is more drought tolerant than S. regelii.

A wheat stay-green mutant, tasg1, was previously generated via mutation breeding of HS2, a common wheat cultivar (Triticum aestivum L.). Compared with wild-type (WT) plants, tasg1 exhibited delayed senescence indicated by the slower degradation of chlorophyll. In this study, the stability of proteins in thylakoid membranes was evaluated in tasg1 under drought stress compared with WT plants in the field as well as in seedlings in the laboratory. Drought stress was imposed by controlling irrigation and sheltering the plants from rain in the field, and by polyethylene glycol (PEG)-6000 in the laboratory. The results indicated that tasg1 plants could maintain higher Hill activity, actual efficiency (ΦPSII), maximal photochemical efficiency of PSII (Fv/Fm), and Ca2+-ATPase and Mg2+-ATPase activities than the WT plants under drought stress. Furthermore, the abundance of some polypeptides in thylakoid membranes of tasg1 was greater than that in the WT under drought stress. Expression levels of TaLhcb4 and TaLhcb6 were higher in tasg1 compared with the WT. Under drought stress, the accumulation of superoxide radical (O2·–) and hydrogen peroxide (H2O2) was lower in tasg1 compared with the WT not only at the senescence stage but also at the seedling stages. These results suggest greater functional stability of thylakoid membrane proteins in tasg1 compared with the WT, and the higher antioxidant competence of tasg1 may play an important role in the enhanced drought tolerance of tasg1.

Deep root hydraulic conductance is upregulated during severe drought and is associated with upregulation in aquaporin activity. In 2011, Texas experienced the worst single-year drought in its recorded history and, based on tree-ring data, likely its worst in the past millennium. In the Edwards Plateau of Texas, rainfall was 58 % lower and the mean daily maximum temperatures were >5 °C higher than long-term means in June through September, resulting in extensive tree mortality. To better understand the balance of deep and shallow root functioning for water supply, we measured root hydraulic conductance (K R) in deep (~20 m) and shallow (5–10 cm) roots of Quercus fusiformis at four time points in the field in 2011. Deep roots of Q. fusiformis obtained water from a perennial underground (18–20 m) stream that was present even during the drought. As the drought progressed, deep root K R increased 2.6-fold from early season values and shallow root K R decreased by 50 % between April and September. Inhibitor studies revealed that aquaporin contribution to K R increased in deep roots and decreased in shallow roots as the drought progressed. Deep root aquaporin activity was upregulated during peak drought, likely driven by increased summer evaporative demand and the need to compensate for declining shallow root K R. A whole-tree hydraulic transport model predicted that trees with greater proportions of deep roots would have as much as five times greater transpiration during drought periods and could sustain transpiration during droughts without experiencing total hydraulic failure. Our results suggest that trees shift their dependence on deep roots versus shallow roots during drought periods, and that upregulation of aquaporin activity accounts for at least part of this increase.

When chilling-sensitive plants are chilled, root hydraulic conductance (L(o)) declines precipitously; L(o) also declines in chilling-tolerant plants, but it subsequently recovers, whereas in chilling-sensitive plants it does not. As a result, the chilling-sensitive plants dry out and may die. Using a chilling-sensitive and a chilling-tolerant maize genotype we investigated the effect of chilling on L(o), and its relationship to osmotic water permeability of isolated root cortex protoplasts, aquaporin gene expression, aquaporin abundance, and aquaporin phosphorylation, hydrogen peroxide (H(2)O(2)) accumulation in the roots and electrolyte leakage from the roots. Because chilling can cause H(2)O(2) accumulation we also determined the effects of a short H(2)O(2) treatment of the roots and examined the same parameters. We conclude from these studies that the recovery of L(o) during chilling in the chilling-tolerant genotype is made possible by avoiding or repairing membrane damage and by a greater abundance and/or activity of aquaporins. The same changes in aquaporins take place in the chilling-sensitive genotype, but we postulate that membrane damage prevents the L(o) recovery. It appears that the aquaporin response is necessary but not sufficient to respond to chilling injury. The plant must also be able to avoid the oxidative damage that accompanies chilling.

In the present work, we have attempted a comprehensive analysis of cytosolic and microsomal proteomes to elucidate the signaling pathways impaired in human hepatoma (Huh7) cells upon herpes simplex virus type 1 (HSV-1; Cgal(+)) infection. Using a combination of differential in-gel electrophoresis and nano liquid chromatography/tandem mass spectrometry, 18 spots corresponding to 16 unique deregulated cellular proteins were unambiguously identified, which were involved in the regulation of essential processes such as apoptosis, mRNA processing, cellular structure and integrity, signal transduction, and endoplasmic-reticulum-associated degradation pathway. Based on our proteomic data and additional functional studies target proteins were identified indicating a late activation of apoptotic pathways in Huh7 cells upon HSV-1 Cgal(+) infection. Additionally to changes on RuvB-like 2 and Bif-1, down-regulation of Erlin-2 suggests stimulation of Ca(2+)-dependent apoptosis. Moreover, activation of the mitochondrial apoptotic pathway results from a time-dependent multi-factorial impairment as inferred from the stepwise characterization of constitutive pro- and anti-apoptotic factors. Activation of serine-threonine protein phosphatase 2A (PP2A) was also found in Huh7 cells upon HSV-1 Cgal(+) infection. In addition, PP2A activation paralleled dephosphorylation and inactivation of downstream mitogen-activated protein (MAP) kinase pathway (MEK(1/2), ERK(1/2)) critical to cell survival and activation of proapoptotic Bad by dephosphorylation of Ser-112. Taken together, our results provide novel molecular information that contributes to define in detail the apoptotic mechanisms triggered by HSV-1 Cgal(+) in the host cell and lead to the implication of PP2A in the transduction of cell death signals and cell survival pathway arrest.

As water and nutrient uptake should be related in the response of plants to salinity, the aim of this paper is to establish whether or not aquaporin functionality is related to H+-ATPase activity in root cells of pepper (Capsicum annuum L.) plants. Thus, H+-ATPase activity was measured in plasma membrane vesicles isolated from roots and aquaporin functionality was measured using a cell pressure probe in intact roots. Salinity was applied as 60 mM NaCl or 60 mM KCl, to determine which ion (Na+, K+ or Cl-) is producing the effects. We also investigated whether the effects of both salts were ameliorated by Ca2+. Similar results were obtained for cell hydraulic conductivity, Lpc, and H+-ATPase activity, large reductions in the presence at NaCl or KCl and an ameliorative effect of Ca2+. However, fusicoccin (an activator of H+-ATPase) did not alter osmotic water permeability of protoplasts isolated from roots. Addition of Hg2+ inhibited both ATPase and aquaporins, but ATPase also contains Hg-binding sites. Therefore, the results indicate that H+-ATPase and aquaporin activities may not be related in pepper plants.

Radial transport of water in excised barley (Hordeum distichon, cv. Villa) roots was measured using a new method based on the pressure-probe technique. After attaching excised roots to the probe, root pressures of 0.9 to 2.9 bar were developed. They could be altered either by changing the root pressure artificially (with the aid of the probe) or by changing the osmotic pressure of the medium in order to induce water flows across the root. The hydraulic conductivity of the barley roots (per cm(2) of outer root surface) was obtained in different types of experiments (initial water flow, pressure relaxations, constant water flow) and was (0.3-4.3)·10(-7) cm s(-1) bar(-1). The hydraulic conductivity of the root was by an order of magnitude smaller than the hydraulic conductivity of the cell membranes of cortical and epidermal cells (0.8-2.2)·10(-6) cm s(-1) bar(-1). The half-times of water exchange of these cells was 1-21 s and two orders of magnitude smaller than that of entire excised roots (100-770 s). Their volumetric elastic modulus was 15-305 bar and increased with increasing turgor. Within the root cortex, turgor was independent of the position of the cell within a certain layer and turgor ranged between 3 and 5 bar. The large difference between the hydraulic conductivity of the root and that of the cell membranes indicates that there is substantial cell-to-cell (transcellular plus symplasmic) transport of water in the root. When it is assumed that 10-12 membrane layers (plasmalemma plus tonoplast) in the epidermis, cortex and endodermis form the hydraulic resistance to water flow, a value for the hydraulic conductivity of the root can be calculated which is similar to the measured value. This picture for water transport in the root contradicts current models which favour apoplasmic water transport in the cortex.

Anthropogenic activities are increasing cadmium (Cd) concentrations in agricultural soils. Cadmium can be absorbed by plant roots and modify the physiology of the plant. Carbon exchange rate (CER) and leaf conductance to water vapor (gs) were measured on the fifth leaf of two soybean (Glycine max [L.] Merr.) cultivars (Illini insensitive and Richland sensitive) for 6 consecutive days; Cd(NO3)2 was added to the hydroponic solution to achieve a final concentration of 50 µmol. At the end of the experiment, stomata length and width, mesophyll limitation to photosynthesis, root hydraulic conductance, relative water content (RWC), and Cd concentration in leaves, stems, and roots were measured on treated and control plants. Cadmium progressively reduced CER and gs to about 50% after 6 d of treatment. This was more evident in Richland than in Illini and was not linked with leaf RWC and mesophyll limitation to photosynthesis. After 6 d, the apparent root hydraulic water conductivity was 67% lower in the Cd‐treated plants than in controls. The primary mechanism affected by Cd‐induced stress in soybean is root water uptake, and this reduction is consistent with the decrease in stomatal opening and conductance, and therefore, in photosynthesis.

The presence of PGP bacteria in the rhizosphere has a positive effect on plant growth and increases their productivity both under favorable and stressful conditions. Despite the fact that much attention is paid to the various mechanisms of action of growth-promoting bacteria on plants, the aspect of the effect of bacteria on changes in the formation of apoplastic barriers and their effect on water transport remains almost unexplored. Previously, it was shown that Pseudomonas mandelii IB-Ki14 accelerated the formation of Casparian bands in wheat plants, although the involvement of LTPs (Lipid Transfer Proteins) in this process has not been shown. The aim of this study was to elucidate the effect of bacterial inoculation on LTP synthesis in the roots of the pea (Pisum sativum) and its relation to the formation of apoplastic barriers and hydraulic conductance. We aimed to test the hypothesis that the decrease in hydraulic conductance due to increased formation of apoplastic barriers can be compensated by increased activity of aquaporin water channels. Activation of genes encoding PsLTP and an increase in the amount of lipid-transporting proteins in pea root phloem induced by P. mandelii IB-Ki14 treatment were found. Inoculation was accompanied by an increase in suberin deposition in the Casparian bands, while hydraulic conductivity did not decrease. The use of Fenton's reagent, which is an inhibitor of aquaporins, showed that the hydraulic conductivity of plants treated with bacteria decreased to a greater extent compared to the control group, which indicates an increase in the activity of aquaporins under the influence of bacteria. We conclude that P. mandelii IB-Ki14 stimulates the deposition of suberin, the biosynthesis of which involves lipid-transporting proteins, and increases the activity of aquaporins, which, in turn, prevents a possible decrease in hydraulic conductivity due to the formation of apoplastic barriers in pea roots.

The arbuscular mycorrhizal (AM) symbiosis has been shown to modulate the same physiological processes as the phytohormone abscisic acid (ABA) and to improve plant tolerance to water deficit. The aim of the present research was to evaluate the combined influence of AM symbiosis and exogenous ABA application on plant root hydraulic properties and on plasma-membrane intrinsic proteins (PIP) aquaporin gene expression and protein accumulation after both a drought and a recovery period. Results obtained showed that the application of exogenous ABA enhanced osmotic root hydraulic conductivity (L) in all plants, regardless of water conditions, and that AM plants showed lower L values than nonAM plants, a difference that was especially accentuated when plants were supplied with exogenous ABA. This effect was clearly correlated with the accumulation pattern of the different PIPs analyzed, since most showed reduced expression and protein levels in AM plants fed with ABA as compared to their nonAM counterparts. The possible involvement of plant PIP aquaporins in the differential regulation of L by ABA in AM and nonAM plants is further discussed.

Protein Implicated in Nonsyndromic Mental Retardation Regulates Protein Kinase A (PKA) Activity

The activation of large conductance Ca 2+ -activated K + (BK) channels results in cellular hyperpolarization and decreased excitation. Protein kinase C (PKC), adenosine 3′,5′-monophosphate (cAMP)-dependent protein kinase (PKA), and guanosine 3′,5′-monophosphate (cGMP)-dependent protein kinase (PKG) regulate the activity of BK channels. However, the mechanism by which these kinases regulate BK channel activity remains largely unknown. Zhou et al. cloned three isoforms of the BK channel α subunit, which differ only in their COOH-termini: both the A (BK A ) and B (BK B ) isoforms have two serine residues that serve as sites for protein kinase C (PKC)-mediated phosphorylation, whereas the C (BK C ) isoform does not. Inside-out cell membrane patches from HEK293 cells that expressed BK A or BK B were activated by the application of PKG but not by PKA, whereas BK C -containing inside-out patches were only activated by the application of PKA. Upon mutation of the PKC-mediated phosphorylation sites (serine to alanine) on BK A , PKA (but not PKG) activated the mutated BK channel in inside-out patches, suggesting that PKC-dependent phosphorylation of BK A switches the sensitivity of the BK channel from PKA to PKG. Mutation of either of the two PKC-mediated phosphorylation sites on BKA reduced, but did not completely eliminate, the capacity of PKG to activate BK A , which suggests that both Ser residues in BK A are important for optimal PKG-dependent activation of the BK channel. Mutation of the known distinct PKA or PKG phosphorylation sites on BK A prevented the activation of BK channels by those kinases, indicating that PKA- or PKG-mediated phosphorylation is essential to BK channel activation. Thus, the data suggest that the activity of PKC determines the sensitivity of BK channel activation to PKA or PKG. X.-B. Zhou, C. Arntz, S. Kamm, K. Motejlek, U. Sausbier, G.-X. Wang, P. Ruth, M. Korth, A molecular switch for specific stimulation of the BK Ca channel by cGMP and cAMP kinase. J. Biol. Chem. 276 , 43239-43245 (2001). [Abstract] [Full Text]

Phosphorylation with protein kinases modulates calcium loading of terminal cisternae of sarcoplasmic reticulum from skeletal muscle