Transpirational Water Flow Research Articles

The outward shaker channel OsK5.2 improves plant salt tolerance by contributing to control of both leaf transpiration and K+ secretion into xylem sap.

Soil salinity constitutes a major environmental constraint to crop production worldwide. Leaf K+ /Na+ homoeostasis, which involves regulation of transpiration, and thus of the xylem sap flow, and control of the ionic composition of the ascending sap, is a key determinant of plant salt tolerance. Here, we show, using a reverse genetics approach, that the outwardly rectifying K+ -selective channel OsK5.2, which is involved in both K+ release from guard cells for stomatal closure in leaves and K+ secretion into the xylem sap in roots, is a strong determinant of rice salt tolerance (plant biomass production and shoot phenotype under saline constraint). OsK5.2 expression was upregulated in shoots from the onset of the saline treatment, and OsK5.2 activity in guard cells led to a fast decrease in transpirational water flow and, therefore, reduced Na+ translocation to shoots. In roots, upon saline treatment, OsK5.2 activity in xylem sap K+ loading was maintained, and even transiently increased, outperforming the negative effect on K+ translocation to shoots resulting from the reduction in xylem sap flow. Thus, the overall activity of OsK5.2 in shoots and roots, which both reduces Na+ translocation to shoots and benefits shoot K+ nutrition, strongly contributes to leaf K+ /Na+ homoeostasis.

The role of NADPH oxidases in regulating leaf gas exchange and ion homeostasis in Arabidopsis plants under cadmium stress

NADPH oxidase, an enzyme associated with the plasma membrane, constitutes one of the main sources of reactive oxygen species (ROS) which regulate different developmental and adaptive responses in plants. In this work, the involvement of NADPH oxidases in the regulation of photosynthesis and cell ionic homeostasis in response to short cadmium exposure was compared between wild type (WT) and three RBOHs (Respiratory Burst Oxidase Homologues) Arabidopsis mutants (AtrbohC, AtrbohD, and AtrbohF). Plants were grown under hydroponic conditions and supplemented with 50 µM CdCl2 for 24 h. Cadmium treatment differentially affected photosynthesis, stomatal conductance, transpiration, and antioxidative responses in WT and Atrbohs mutants. The loss of function of RBOH isoforms resulted in higher Cd2+ influx, mainly in the elongation zone of roots, which was more evident in AtrbohD and AtrbohF mutants. In the mature zone, the highest Cd2+ influx was observed in rbohC mutant. The lack of functional RBOH isoforms also resulted in altered patterns of net K+ transport across cellular membranes, both in the root epidermis and leaf mesophyll. The analysis of expression of metal transporters by qPCR demonstrated that a loss of functional RBOH isoforms has altered transcript levels for metal NRAMP3, NRAMP6 and IRT1 and the K+ transporters outward-rectifying K+ efflux GORK channel, while RBOHD specifically regulated transcripts for high-affinity K+ transporters KUP8 and HAK5, and IRT1 and RBOHD and F regulated the transcription factors TGA3 and TGA10. It is concluded that RBOH-dependent H2O2 regulation of ion homeostasis and Cd is a highly complex process involving multilevel regulation from transpirational water flow to transcriptional and posttranslational modifications of K/metals transporters.

Root and cell hydraulic conductivity, apoplastic barriers and aquaporin gene expression in barley (Hordeum vulgare L.) grown with low supply of potassium.

Limited supply of mineral nutrients often reduces plant growth and transpirational water flow while increasing the ratio of water-absorbing root to water-losing shoot surface. This could potentially lead to an imbalance between water uptake (too much) and water loss (too little). The aim of the present study was to test whether, as a countermeasure, the hydraulic properties (hydraulic conductivity, Lp) of roots decrease at organ and cell level and whether any decreases in Lp are accompanied by decreases in the gene expression level of aquaporins (AQPs) or increases in apoplastic barriers to radial water movement. Barley plants were grown hydroponically on complete nutrient solution, containing 2 mm K+ (100 %), or on low-K solution (0.05 mm K+; 2.5 %), and analysed when they were 15-18 d old. Transpiration, fresh weight, surface area, shoot water potential (ψ), K and Ca concentrations, root (exudation) and cortex cell Lp (cell pressure probe), root anatomy (cross-sections) and AQP gene expression (qPCR) were analysed. The surface area ratio of root to shoot increased significantly in response to low K. This was accompanied by a small decrease in the rate of water loss per unit shoot surface area, but a large (~50 %) and significant decrease in Lp at root and cortex cell levels. Aquaporin gene expression in roots did not change significantly, due to some considerable batch-to-batch variation in expression response, though HvPIP2;5 expression decreased on average by almost 50 %. Apoplastic barriers in the endodermis did not increase in response to low K. Barley plants that are exposed to low K adjust to an increased ratio of root (water uptake) to shoot (water loss) surface primarily through a decrease in root and cell Lp. Reduced gene expression of HvPIP2;5 may contribute to the decrease in Lp.

The significance of water co-transport for sustaining transpirational water flow in plants: a quantitative approach.

In a recent Opinion paper, Wegner (Journal of Experimental Botany 65, 381-392, 2014) adapts a concept developed for water flow in animal tissues to propose a model, which can explain the loading of water into the root xylem against a difference in water potential (Ψ) between the xylem parenchyma cell (more negative Ψ) and the xylem vessel (less negative Ψ). In this model, the transport of water is energized through the co-transport of ions such as K(+) and Cl(-) through plasma membrane-located transporters. The emphasis of the model is on the thermodynamic feasibility of the co-transport mechanism per se. However, what is lacking is a quantitative evaluation of the energy input required at the organismal level to sustain such a co-transport mechanism in the face of considerable net (transpirational) flows of water through the system. Here, we use a ratio of 500 water molecules being co-transported for every pair of K(+) and Cl(-) ions, as proposed for the animal system, to calculate the energy required to sustain daytime and night-time transpirational water flow in barley plants through a water co-transport mechanism. We compare this energy with the total daily net input of energy through photosynthetic carbon assimilation. Water co-transport can facilitate the filling of xylem against a difference in Ψ of 1.0MPa and puts a minor drain on the energy budget of the plant. Based on these findings it cannot be excluded that water co-transport in plants contributes significantly to xylem filling during night-time and possibly also daytime transpiration.

A thermodynamic analysis of the feasibility of water secretion into xylem vessels against a water potential gradient

A series of recent publications has launched a debate on trans-membrane water secretion into root xylem vessels against a water potential gradient, energised by a cotransport with salts (e.g. KCl) that follow their chemical potential gradient. Cation-chloride-cotransporter -type transporters that function in this way in mammalian epithelia were detected in root stelar cells bordering on xylem vessels. Using literature data on barley (Hordeum vulgare L.) seedlings, one study confirmed that K+ and Cl- gradients across stelar cell membranes favour salt efflux. Moreover, the energetic costs of putative water secretion into the xylem (required for maintaining ionic gradients) would amount to just 0.12% of the energy captured by photosynthetic C assimilation if transpirational water flow relied exclusively on this mechanism. Here, a detailed thermodynamic analysis of water secretion into xylem vessels is undertaken, including an approach that exploits its analogy to a desalinisation process. Water backflow due to the passive hydraulic conductivity of stelar cell membranes is also considered. By comparing free energy consumption by putative water secretion with (i) the free energy pool provided by root respiration and (ii) stelar ATPase activity, the feasibility of this mechanism is confirmed but is shown to depend critically on the plant's energy status.

Plant morphophysiological and anatomical factors associated with nitrous oxide flux from wheat (Triticum aestivum)

Experiments were conducted to study the dynamics of nitrous oxide (N₂O) emission from wheat varieties viz., Sonalika, HUW 468, HUW 234 and DBW 14 grown in alluvial soils of North Bank Plain Agroclimatic Zone of Assam, India. Attempts were made to find out the relationship of N₂O emission with plant morphophysiological, anatomical and soil properties. N₂O fluxes from wheat varieties ranged from 40 μg N₂O-N m⁻² h⁻¹ to 295 μg N₂O-N m⁻² h⁻¹. Soil organic carbon and soil temperature have shown significant relationship with N₂O flux. The rate of leaf transpiration recorded from the wheat varieties at different growth stages exhibited a positive correlation with N₂O emission suggesting that movement of N₂O along with the transpirational water flow may be an important mechanism of N₂O transport and emission through wheat plants. Anatomical investigation by scanning electron microscope revealed that N₂O emission has relationship with stomatal frequency of leaf and leaf sheaths. Variety HUW 234 with the highest stomatal frequency of leaf and leaf sheath also recorded higher seasonal N₂O emission compared to other varieties. Seasonal N₂O emission (E(sif)) of the varieties ranged from 3.25 to 3.81 kg N₂O-N ha⁻¹. Significant variations in E(sif) values were recorded within the varieties.

Effects of sodium nitroprusside, the nitric oxide donor, on photosynthesis and ultrastructure of common flax leaf blades

Solutions of sodium nitroprusside, a nitric oxide donor, were introduced at various concentrations into common flax (Linum usitatissimum L.) shoots with the transpirational water flow. Sodium nitroprusside and nitrate were found to exert similar effects on incorporation of 14C into photosynthetic products, leaf cell ultrastructure, and the export of assimilates from leaves. The results suggest that export of assimilates from leaves might be regulated by the products of incomplete nitrate reduction and that regulation may involve the NO-signaling system.

Water Dynamics in Mango (Mangifera indica L.) Fruit during the Young and Mature Fruit Seasons as Measured by the Stem Heat Balance Method

Water flows in the stem and peduncle of mango fruit were monitored, and the surface transpiration from the fruit was measured during the fruit-growing season. The stem heat balance method was used on the peduncle to monitor the inward water flow during the nighttime, and the reverse water flow from the fruit during the daytime when the stem transpiration water flow increased. This diurnal fluctuation pattern in the water flow was more evident in mature fruit than in young fruit. In mature fruit, the daily water loss due to the reverse flow was estimated to be 3% of the fruit weight. The reverse flow water loss and transpired water loss were compensated for by nocturnal inward water flow, through the peduncle, of 30 g over 15 h. These results were well supported by measurements of fruit dimensions, which indicated a circadian rhythm of contraction and expansion. The reverse flow amounted to 80% of the water loss from the daytime contraction of the fruit, a much greater proportion than the fruit surface transpiration.

Water uptake by roots of maize and sunflower affects the radial transport of abscisic acid and its concentration in the xylem

The radial movement of cis-abscisic acid (ABA) has been investigated in young excised roots of Zea mays L. and Helianthus annuus L. which were grown hydroponically. In addition to the symplastic path, ABA was largely translocated across the root apoplast by solvent drag with the water in the transpiration stream. On the apoplastic path ABA may even cross the endodermis. Depending on the ABA concentration of the medium (range: 5–500 nM) and in the root apoplast, the solvent-drag component of the flow of ABA counteracted the dilution of ABA in the xylem caused by transpirational water flow. Acidification of the rhizosphere and of the root apoplast increased the apoplastic transport component. In sunflower, the apoplastic flow of ABA was significantly weaker than in maize roots. This was also indicated by the larger apparent reflection coefficient (σABA) of sunflower roots for ABA (sunflower: σABA = 0.97 ± 0.02, n = 6 roots; maize: σABA = 0.68 ± 0.06, n = 6 roots; ±SD). For both species, σABA was smaller than unity. Root reflection coefficients were affected by factors such as pH, ABA concentration of the medium, and by the suction force applied to excised root systems. Due to the complex composite structure of the permeation barrier in the root, the reflection coefficient estimated from solvent drag is also complex. Since unstirred layers affected the absolute value of the reflection coefficient, σABA has been termed `apparent'. It is concluded that the pH and ABA concentration of the soil solution as well as the transpiration rate (suction force) modify the intensity of the root-to-shoot signal which is influenced by an apoplastic bypass flow of ABA. The latter may be substantially affected by the existence of Casparian bands in the exodermis, which were lacking in the roots studied in this paper.

Growth of Zea mays L. plants with their seminal roots only. Effects on plant development, xylem transport, mineral nutrition and the flow and distribution of abscisic acid (ABA) as a possible shoot to root signal

Maize (Zea mays L.) was grown in quartz sand culture either with a normal root system (controls) or with seminal roots only ('single-rooted'). Development of adventitious roots was prevented by using plants with an etiolated mesocotyl and the stem base was positioned 5-8 cm above the sand. Even though the roots of the single-rooted plants were sufficiently supplied with water and nutrients, the leaves experienced water deficits and showed decreased transpiration as transpirational water flow was restricted by the constant number of xylem vessels present in the mesocotyl. As a consequence of this restriction, transpirational water flow velocities in the metaxylem vessels reached mean values of 270 m h -1 and phloem transport velocities of 5.2 m h -1 . Despite limited xylem transport mineral nutrient concentrations in leaf tissues were not decreased in single-rooted plants, but shoot and particularly stem development was somewhat inhibited. Due to the lack of adventitious roots the shoot:root ratio was strongly increased in the single-rooted plants, but the seminal roots showed compensatory growth compared to those in control plants. Consistent with decreased leaf conductance, ABA concentrations in leaves of single-rooted plants were elevated up to 10-fold, but xylem sap ABA concentrations in these plants were lower than in controls, in good agreement with the well-watered conditions experienced by the seminal roots. Surprisingly, however, ABA concentrations in tissues of the seminal roots of the single-rooted plants were clearly increased compared to the controls, presumably due to increased ABA import via phloem from the water-stressed leaves. The results are discussed in relation to the role of ABA as a shoot to root signal.

Physiological Responses of Red Maple Saplings To Sub-Irrigation With an Untreated Municipal Landfill Leachate

An experiment was undertaken to examine the response of hydroponically-grown red maple ( Acer rubrum L.) saplings to a series of four flooding (sub-irrigation) treatments distributed over a 25-day period with an untreated (saline) municipal solid waste landfill leachate or deionized water. Net photosynthesis rates measured for water-treated saplings rapidly declined to 62% of the levels measured in untreated (control) saplings, but returned to pre-treatment levels with subsequent flooding treatments. Net photosynthesis rates measured for leachate-treated saplings decreased to about 50% of the levels measured for control saplings over the 25-day treatment period, and remained suppressed. Loss of turgor in leaves and a iron-oxyhydroxide plaque on root surfaces were also observed. Reasons proposed for this acute and apparently irreversible response to leachate exposure include: (i) extreme root anaerobiosis conditions caused by root system flooding and exacerbated by a high chemical oxygen demand leachate; (ii) increased root-soil interface resistance to transpiration water flow (osmotic potential gradient, iron oxyhydroxide plaque); (iii) metabolic intolerance to high solute concentrations in plant tissue; and (iv) exposure to potentially toxic volatile organic compounds. Water sub-irrigation had virtually no effect on nutrient and non-nutrient element concentrations in foliage or on the spectral reflectance characteristics of the leaves. Leachate treatment decreased the foliar content of many plant macro- and micro-nutrients significantly, and shifts in spectral reflectance patterns indicated declining plant vigour. Certain chemical constituents present in high concentrations in the leachate irrigant and which can be phytotoxic, such as Cl, accumulated to a significant degree in leachate-treated plant tissue.

Physiological responses of red maple saplings to sub-irrigation with an untreated municipal landfill leachate

An experiment was undertaken to examine the response of hydroponically-grown red maple ( Acer rubrum L.) saplings to a series of four flooding (sub-irrigation) treatments distributed over a 25-day period with an untreated (saline) municipal solid waste landfill leachate or deionized water. Net photosynthesis rates measured for water-treated saplings rapidly declined to 62% of the levels measured in untreated (control) saplings, but returned to pre-treatment levels with subsequent flooding treatments. Net photosynthesis rates measured for leachate-treated saplings decreased to about 50% of the levels measured for control saplings over the 25-day treatment period, and remained suppressed. Loss of turgor in leaves and a iron-oxyhydroxide plaque on root surfaces were also observed. Reasons proposed for this acute and apparently irreversible response to leachate exposure include: (i) extreme root anaerobiosis conditions caused by root system flooding and exacerbated by a high chemical oxygen demand leachate; (ii) increased root-soil interface resistance to transpiration water flow (osmotic potential gradient, iron oxyhydroxide plaque); (iii) metabolic intolerance to high solute concentrations in plant tissue; and (iv) exposure to potentially toxic volatile organic compounds. Water sub-irrigation had virtually no effect on nutrient and non-nutrient element concentrations in foliage or on the spectral reflectance characteristics of the leaves. Leachate treatment decreased the foliar content of many plant macro- and micro-nutrients significantly, and shifts in spectral reflectance patterns indicated declining plant vigour. Certain chemical constituents present in high concentrations in the leachate irrigant and which can be phytotoxic, such as Cl, accumulated to a significant degree in leachate-treated plant tissue.

Change in Water Conducting Properties of Plant Roots by Nutrition and Salt Stress

Mung bean plants were grown in half strength Hoagland nutrient solution and with only 2 mM N and no P. One set of these plants was exposed to 25 mM NaCl salinity. Plant growth was depressed to different degrees with the nutrition and salt stress. Measurements on the volume flow (Jv) and hydraulic conductivity (Lp) of the detopped 23 day old plants were made under applied pressure of 0.207 MPa. The transpirational water flow (Jt) and the values of Jv were similar for most of the treatments. Values of Jt, Jv and Lp of plants decreased when both the nutrition and the salinity stress were imposed. Simultaneous stress of nutrients and salinity appeared to damage the cell membranes.