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Magnetic resonance microscopy of changes in water content in stems of transpiring plants.

Differences in water content and degree of binding in the various stem tissues of Pelargonium hortorum were observed by magnetic resonance imaging. 1H images were obtained with a resolution of 100 microns in the transverse plane and a slice thickness of 1250 microns. It was possible to distinguish the principal tissues of the stem by differences in their proton density or apparent water content and spin lattice relaxation time (T1) or degree of water binding. Measurements were made while the plant was slowly and actively transpiring. In the slowly transpiring plant, T1 of various tissues ranged from an average of 659 to 865 ms with a proton density variation of from 72 to 100%. In the actively transpiring plant, T1 ranged from an average of 511 to 736 ms, and the proton density was reduced, ranging between 62 and 88% of the peak value found in the slowly transpiring plant. The fibrous sheath surrounding the vascular tissue and the epidermal region was found to have the highest spin density and T1. Both tissues are comprised of relatively small thick-walled cells. Cortical and pith parenchyma are composed of larger, thinner-walled cells with numerous intercellular spaces and lower spin density and T1. The differences are attributed to the higher water content by volume in the tissue composed of smaller, more compactly arranged cells. The resolution obtained in this work enables clear definition of tissues in the living plant and quantitative information concerning differences in the distribution and extent of binding of water.

Magnetic Resonance Spectroscopy↗

The ERECTA gene regulates plant transpiration efficiency in Arabidopsis.

Assimilation of carbon by plants incurs water costs. In the many parts of the world where water is in short supply, plant transpiration efficiency, the ratio of carbon fixation to water loss, is critical to plant survival, crop yield and vegetation dynamics. When challenged by variations in their environment, plants often seem to coordinate photosynthesis and transpiration, but significant genetic variation in transpiration efficiency has been identified both between and within species. This has allowed plant breeders to develop effective selection programmes for the improved transpiration efficiency of crops, after it was demonstrated that carbon isotopic discrimination, Delta, of plant matter was a reliable and sensitive marker negatively related to variation in transpiration efficiency. However, little is known of the genetic controls of transpiration efficiency. Here we report the isolation of a gene that regulates transpiration efficiency, ERECTA. We show that ERECTA, a putative leucine-rich repeat receptor-like kinase (LRR-RLK) known for its effects on inflorescence development, is a major contributor to a locus for Delta on Arabidopsis chromosome 2. Mechanisms include, but are not limited to, effects on stomatal density, epidermal cell expansion, mesophyll cell proliferation and cell-cell contact.

Arabidopsis↗

The Arabidopsis outward K+ channel GORK is involved in regulation of stomatal movements and plant transpiration.

Microscopic pores present in the epidermis of plant aerial organs, called stomata, allow gas exchanges between the inner photosynthetic tissue and the atmosphere. Regulation of stomatal aperture, preventing excess transpirational vapor loss, relies on turgor changes of two highly differentiated epidermal cells surrounding the pore, the guard cells. Increased guard cell turgor due to increased solute accumulation results in stomatal opening, whereas decreased guard cell turgor due to decreased solute accumulation results in stomatal closing. Here we provide direct evidence, based on reverse genetics approaches, that the Arabidopsis GORK Shaker gene encodes the major voltage-gated outwardly rectifying K(+) channel of the guard cell membrane. Expression of GORK dominant negative mutant polypeptides in transgenic Arabidopsis was found to strongly reduce outwardly rectifying K(+) channel activity in the guard cell membrane, and disruption of the GORK gene (T-DNA insertion knockout mutant) fully suppressed this activity. Bioassays on epidermal peels revealed that disruption of GORK activity resulted in impaired stomatal closure in response to darkness or the stress hormone abscisic acid [corrected]. Transpiration measurements on excised rosettes and intact plants (grown in hydroponic conditions or submitted to water stress) revealed that absence of GORK activity resulted in increased water consumption. The whole set of data indicates that GORK is likely to play a crucial role in adaptation to drought in fluctuating environments.

Arabidopsis↗

Estimation of whole-plant transpiration of bananas using sap flow measurements.

Banana, one of the largest rhizomatous herbs in the world, is the fourth most important global food crop. It has a high water requirement, but the whole-plant water use in the field has not been determined satisfactorily. In this study, whole-plant water use in potted and field-grown banana plants (Musa 'Cavendish' cv. Williams) was successfully determined using a xylem sap flow method. This was achieved using Granier sensor probes implanted into the central cylinder of the banana corm. The whole-plant water use in field-grown bananas was 9-10 l plant(-1) d(-1). The values of daily total sap flow in potted plants correlated closely with gravimetric measurements (r(2)=0.92) and with changes in soil water status (r(2)=0.77). In well-watered, mature, field-grown plants, hourly sap flow also closely correlated with changes in solar radiation, vapour pressure deficit and evapotranspiration. The study indicates that sap flow measurement is a sensitive and accurate method for determining whole-plant water use in bananas under potted as well as field conditions.

Algorithms↗

Responses of gas exchange to reversible changes in whole-plant transpiration rate in two conifer species.

This study examined the autonomy of branches with respect to the control of transpiration (E) in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and western red cedar (Thuja plicata Donn) seedlings. Experiments were conducted on whole seedlings in a gas exchange system with a dual-cuvette that permitted independent manipulation and measurement of E in the upper and lower cuvettes. The value of E in one cuvette was manipulated by varying vapor pressure deficit (D) between 2.2 and 0.2 kPa, whereas D in the other cuvette was held at 2.2 kPa. Reducing D, while increasing stomatal conductance (gs), resulted in an overall decrease in E. In western red cedar, this decrease was almost threefold, and in Douglas-fir, approximately fourfold. In well-watered western red cedar, a reduction of whole-plant E by 46% (brought about by reducing D in the upper cuvette) resulted in a 12% increase in gs, a 12% increase in E and a 7% increase in net assimilation (A) of untreated foliage in the lower cuvette. Responses of gs, E and A of untreated foliage were similar irrespective of whether foliage was at the top or bottom of the seedling. When D in the treatment cuvette was restored to 2.2 kPa, gs, E and A of foliage in the untreated cuvette returned to pretreatment values. In contrast, in well-watered Douglas-fir, there was almost no change in gs, E or A of untreated foliage in one cuvette when D in the other cuvette was reduced, causing a 52% reduction in whole-plant E. However, similar manipulations on drought-stressed Douglas-fir led to 7-19% increases in gs, E and A of untreated foliage. In well-watered western red cedar, daytime leaf water potential (Psil) was maintained near -0.9 MPa over a wide range of D, whereas Psil of Douglas-fir decreased from -1.2 to -1.5 MPa as D increased. The tighter (isohydric) regulation of Psil in western red cedar may partly explain its greater stomatal response to D and variation in whole-plant E compared with Douglas-fir. In response to a reduction in E, measured increases in Psil and gs of unmanipulated foliage were less than predicted by a model assuming complete hydraulic connectivity of foliage. Our results suggest the foliage of both species is partially autonomous with respect to water.

Photosynthesis↗

Diurnal variations in hydraulic conductivity and root pressure can be correlated with the expression of putative aquaporins in the roots of lotus japonicus

The hydraulic conductivity of excised roots (Lp(r)) of the legume Lotus japonicus (Regel) K. Larsen grown in mist (aeroponic) and sand cultures, was found to vary over a 5-fold range during a day/night cycle. This behaviour was seen when Lp(r) was measured in roots exuding, either under root pressure (osmotic driving force), or under an applied hydrostatic pressure of 0.4 MPa which produced a rate of water flow similar to that in a transpiring plant. A similar daily pattern of variation was seen in plants grown in natural daylight or in controlled-environment rooms, in plants transpiring at ambient rates or at greatly reduced rates, and in plants grown in either aeroponic or sand culture. When detached root systems were connected to a root pressure probe, a marked diurnal variation was seen in the root pressure generated. After excision, this circadian rhythm continued for some days. The hydraulic conductivity of the plasma membrane of individual root cells was measured during the diurnal cycle using a cell pressure probe. Measurements were made on the first four cell layers of the cortex, but no evidence of any diurnal fluctuation could be found. It was concluded that the conductance of membranes of endodermal and stelar cells may be responsible for the observed diurnal rhythm in root Lp(r). When mRNAs from roots were probed with cDNA from the Arabidopsis aquaporin AthPIP1a gene, an abundant transcript was found to vary in abundance diurnally under high-stringency conditions. The pattern of fluctuations resembled closely the diurnal pattern of variation in root Lp(r). The plasma membranes of root cells were found to contain an abundant hydrophobic protein with a molecular weight of about 31 kDa which cross-reacted strongly to an antibody raised against the evolutionarily conserved N-terminal amino acid sequence of AthPIP1a.

Journal Article↗

Genotype-dependent variation in the transpiration efficiency of plants and photosynthetic activity of flag leaves in spring barley under varied nutrition.

In the study, spring barley genotypes of various origin and breeding history were found to show a broad genetic variation in the vegetative and generative measures of the whole-plant transpiration efficiency (TE), photosynthesis (A) and transpiration (E) rates of flag leaves, leaf efficiency of gas exchange (A/E) and stress tolerance (T) when grown till maturity in soil-pots under high and reduced NPK supplies. Broad-sense heritabilities for the characteristics ranged from 0.61 to 0.87. Significant genotype-nutrition interactions were noticed, constituting 19-23% of the total variance in TE measures. The results suggest that at least some 'exotic' accessions from Ethiopia, Syria, Morocco and/or Tibet may serve as attractive genetic sources of novel variations in TE, T and A for the breeding of barleys of improved adaptation to less favourable fertilisation.

Analysis of Variance↗

Element concentrations in the xylem sap of Picea abies (L.) Karst. seedlings extracted by various methods under different environmental conditions.

We used a Scholander pressure chamber to assess the effects of various extraction methods under different environmental conditions on element concentrations in xylem sap of 3-year-old Picea abies (L.) Karst. seedlings. Sap from excised shoots contained higher element concentrations when extracted at low than at high over-pressures. When comparing plants differing in water status, we found that a high extraction over-pressure introduced a systematic error into the data. For example, in well-watered non-transpiring plants relative to unwatered transpiring plants, potassium concentrations were 70% higher in sap extracted at 0.1 MPa over-pressure, but only 10% higher in sap extracted at 1.0 MPa over-pressure. Moreover, treatment effects depended on the time of day when the sap was extracted. Increased water flux in transpiring plants relative to non-transpiring plants resulted in reduced xylem sap element concentrations when samples were collected after 9 h of transpiration, but not after 4 to 6 h of transpiration. Drought had little effect on xylem sap element concentrations, indicating that rates of element release into xylem conduits, element depletion by growing tissues, and water flow maintained a balance that may prevent nutrient stress during short-term drought.

Journal Article↗

Characterizing photosynthesis and transpiration of plant communities in controlled environments.

CO2 and water vapor fluxes of hydroponically grown wheat and soybean canopies were measured continuously in several environments with an open gas exchange system. Canopy CO2 fluxes reflect the photosynthetic efficiency of a plant community, and provide a record of plant growth and health. There were significant diurnal fluctuations in root and shoot CO2 fluxes, and in shoot water vapor fluxes. Canopy stomatal conductance (Gc) to water vapor was calculated from simultaneous measurements of canopy temperature (Tcan) and transpiration rates (Tr). Tr in the dark was substantial, and there were large diurnal fluctuations in both Gc and Tr. Canopy net Photosynthesis (Pnet), Tr, and Gc increased with increasing net radiation. Gc increased with Tr, suggesting that the stomata of plants in controlled environments (CEs) behave differently from field-grown plants. A transpiration model based on measurements of Gc was developed for CEs. The model accurately predicted Tr from a soybean canopy.

Carbon Dioxide↗

Unraveling the effects of plant hydraulics on stomatal closure during water stress in walnut.

The objectives of the study were to identify the relevant hydraulic parameters associated with stomatal regulation during water stress and to test the hypothesis of a stomatal control of xylem embolism in walnut (Juglans regia x nigra) trees. The hydraulic characteristics of the sap pathway were experimentally altered with different methods to alter plant transpiration (Eplant) and stomatal conductance (gs). Potted trees were exposed to a soil water depletion to alter soil water potential (Psisoil), soil resistance (Rsoil), and root hydraulic resistances (Rroot). Soil temperature was changed to alter Rroot alone. Embolism was created in the trunk to increase shoot resistance (Rshoot). Stomata closed in response to these stresses with the effect of maintaining the water pressure in the leaf rachis xylem (P(rachis)) above -1.4 MPa and the leaf water potential (Psileaf) above -1.6 MPa. The same dependence of Eplant and gs on P(rachis) or Psileaf was always observed. This suggested that stomata were not responding to changes in Psisoil, Rsoil, Rroot, or Rshoot per se but rather to their impact on P(rachis) and/or Psileaf. Leaf rachis was the most vulnerable organ, with a threshold P(rachis) for embolism induction of -1.4 MPa. The minimum Psileaf values corresponded to leaf turgor loss point. This suggested that stomata are responding to leaf water status as determined by transpiration rate and plant hydraulics and that P(rachis) might be the physiological parameter regulated by stomatal closure during water stress, which would have the effect of preventing extensive developments of cavitation during water stress.

Acclimatization↗

Effects of phenology, water availability and seed source on loblolly pine biomass partitioning and transpiration.

One-year-old loblolly pine (Pinus taeda L.) seedlings from four seed sources (Arkansas, Georgia, Texas and Virginia) grown in 1-m-deep sand-filled pits in two water regimes (well-watered and drought) were studied, to gain insight into the process of seedling establishment. Whole-plant transpiration was measured biweekly from July to December. Whole-plant harvests were conducted at 6-week intervals from April to December. Whole-plant transpiration and transpiration per unit leaf and root area were affected by treatment, seedlot and phenology. Seedlings of the Arkansas seedlot maintained significantly higher transpiration rates per unit leaf and root area during drought than seedlings of the Virginia, Georgia or Texas seedlots, but did not accumulate greater biomass. The high transpiration rates of the Arkansas seedlings were attributed to their deep root systems. Allometric relationships indicated that, relative to the whole plant, biomass allocation to needles of drought-treated seedlings was enhanced during the summer (allometric ratio 1.09), whereas allocation to roots was enhanced in the spring and fall (allometric ratios of 1.13 and 1.09, respectively). Relative to the whole plant, biomass allocation to needles of well-watered seedlings was enhanced throughout the experiment (allometric ratio of 1.16 declining to 1.05), whereas the allometric ratio of root to total biomass was 0.89 or less throughout. Allometric relationships also indicated variation in biomass partitioning to roots in three soil layers (0-30, 30-60 and 60-100 cm), which differed among harvests in each soil layer. Root growth in both well-watered and drought-treated seedlings was concentrated in the top soil layer in the spring, shifted to the middle and bottom soil layers in the summer, and then increased in the top soil layer in the fall. Compared with well-watered seedlings, drought-treated seedlings had higher rates of root growth in the bottom soil layer in the fall, a characteristic that would confer tolerance to future periods of limited soil water availability.

Biomass↗

Control of Leaf Expansion Rate of Droughted Maize Plants under Fluctuating Evaporative Demand (A Superposition of Hydraulic and Chemical Messages?).

We have analyzed the possibility that chemical signaling does not entirely account for the effect of water deficit on the maize (Zea mays L.) leaf elongation rate (LER) under high evaporative demand. We followed time courses of LER (0.2-h interval) and spatial distribution of elongation rate in leaves of either water-deficient or abscisic acid (ABA)-fed plants subjected to varying transpiration rates in the field, in the greenhouse, and in the growth chamber. At low transpiration rates the effect of the soil water status on LER was related to the concentration of ABA in the xylem sap and could be mimicked by feeding artificial ABA. Transpiring plants experienced a further reduction in LER, directly linked to the transpiration rate or leaf water status. Leaf zones located at more than 20 mm from the ligule stopped expanding during the day and renewed expansion during the night. Neither ABA concentration in the xylem sap, which did not appreciably vary during the day, nor ABA flux into shoots could account for the effect of evaporative demand. In particular, maximum LER was observed simultaneously with a minimum ABA flux in the droughted plants, but with a maximum ABA flux in ABA-fed plants. All data were interpreted as the superposition of two additive effects: the first involved ABA signaling and was observed during the night and in ABA-fed plants, and the second involved the transpiration rate and was observed even in well-watered plants. We suggest that a hydraulic signal is the most likely candidate for this second effect.

Journal Article↗