Ultrastructure of Plant Leaf Cuticles in relation to Sample Preparation as Observed by Transmission Electron Microscopy

The leaf cuticular ultrastructure of some plant species has been examined by transmission electron microscopy (TEM) in only few studies. Attending to the different cuticle layers and inner structure, plant cuticles have been grouped into six general morphological types. With the aim of critically examining the effect of cuticle isolation and preparation for TEM analysis on cuticular ultrastructure, adaxial leaf cuticles of blue-gum eucalypt, grey poplar, and European pear were assessed, following a membrane science approach. The embedding and staining protocols affected the ultrastructure of the cuticles analysed. The solubility parameter, surface tension, and contact angles with water of pure Spurr's and LR-White resins were within a similar range. Differences were however estimated for resin : solvent mixtures, since Spurr’s resin is combined with acetone and LR-White resin is mixed with ethanol. Given the composite hydrophilic and lipophilic nature of plant cuticles, the particular TEM tissue embedding and staining procedures employed may affect sample ultrastructure and the interpretation of the results in physicochemical and biological terms. It is concluded that tissue preparation procedures may be optimised to facilitate the observation of the micro- and nanostructure of cuticular layers and components with different degrees of polarity and hydrophobicity.

Wettability, polarity and water absorption of Quercus ilex leaves: effect of leaf side and age

Plant trichomes play important protective functions and may have a major influence on leaf surface wettability. With the aim of gaining insight into trichome structure, composition and function in relation to water-plant surface interactions, we analyzed the adaxial and abaxial leaf surface of Quercus ilex L. (holm oak) as model. By measuring the leaf water potential 24 h after the deposition of water drops on to abaxial and adaxial surfaces, evidence for water penetration through the upper leaf side was gained in young and mature leaves. The structure and chemical composition of the abaxial (always present) and adaxial (occurring only in young leaves) trichomes were analyzed by various microscopic and analytical procedures. The adaxial surfaces were wettable and had a high degree of water drop adhesion in contrast to the highly unwettable and water repellent abaxial holm oak leaf sides. The surface free energy, polarity and solubility parameter decreased with leaf age, with generally higher values determined for the abaxial sides. All holm oak leaf trichomes were covered with a cuticle. The abaxial trichomes were composed of 8% soluble waxes, 49% cutin, and 43% polysaccharides. For the adaxial side, it is concluded that trichomes and the scars after trichome shedding contribute to water uptake, while the abaxial leaf side is highly hydrophobic due to its high degree of pubescence and different trichome structure, composition and density. Results are interpreted in terms of water-plant surface interactions, plant surface physical-chemistry, and plant ecophysiology.

Implications of the mesophyll conductance to CO2 for photosynthesis and water-use efficiency during long-term water stress and recovery in two contrasting Eucalyptu species

Water stress (WS) slows growth and photosynthesis (An), but most knowledge comes from short-time studies that do not account for longer term acclimation processes that are especially relevant in tree species. Using two Eucalyptus species that contrast in drought tolerance, we induced moderate and severe water deficits by withholding water until stomatal conductance (gsw) decreased to two pre-defined values for 24 d, WS was maintained at the target gsw for 29 d and then plants were re-watered. Additionally, we developed new equations to simulate the effect on mesophyll conductance (gm) of accounting for the resistance to refixation of CO2. The diffusive limitations to CO2, dominated by the stomata, were the most important constraints to An. Full recovery of An was reached after re-watering, characterized by quick recovery of gm and even higher biochemical capacity, in contrast to the slower recovery of gsw. The acclimation to long-term WS led to decreased mesophyll and biochemical limitations, in contrast to studies in which stress was imposed more rapidly. Finally, we provide evidence that higher gm under WS contributes to higher intrinsic water-use efficiency (iWUE) and reduces the leaf oxidative stress, highlighting the importance of gm as a target for breeding/genetic engineering.

Stability Analysis of a Run-of-River Diversion Hydropower Plant With Surge Tank And Spillway in the Head Pond

Run-of-river hydropower plants usually lack significant storage capacity; therefore, the more adequate control strategy would consist of keeping a constant water level at the intake pond in order to harness the maximum amount of energy from the river flow or to reduce the surface flooded in the head pond. In this paper, a standard PI control system of a run-of-river diversion hydropower plant with surge tank and a spillway in the head pond that evacuates part of the river flow plant is studied. A stability analysis based on the Routh-Hurwitz criterion is carried out and a practical criterion for tuning the gains of the PI controller is proposed. Conclusions about the head pond and surge tank areas are drawn from the stability analysis. Finally, this criterion is applied to a real hydropower plant in design state; the importance of considering the spillway dimensions and turbine characteristic curves for adequate tuning of the controller gains is highlighted

Potential issues related to emergency core cooling system strainers performance at boiling water reactors: application to Cofrentes NPP (Spain)

From the 60s to the 90s, a great number of events related to the Emergency Core Cooling Systems Strainers have been happened in all kind of reactors all over the world. Thus, the Nuclear Regulatory Commission of the USA emitted some Bulletins to address the concerns about the adequacy of Emergency Core Cooling Systems (ECCS) strainer performance at boiling water reactors (BWR). In Spain the regulatory body (Consejo de Seguridad Nuclear, CSN) adopted the USA regulation and Cofrentes NPP installed new strainers with a considerable bigger size than the old strainers. The nuclear industry conducted significant and extensive research, guidance development, testing, reviews, and hardware and procedure changes during the 90s to resolve the issues related to debris blockage of BWR strainers. In 2001 the NRC and CSN closed the Bulletins. Thereafter, the strainers issues were moved to the PWR reactors. In 2004 the NRC issued a Generic Letter (GL). It requested the resolution of several effects which were not noted in the past. The GL regarded to be resolved by the PWR reactors but the NRC in USA and the CSN in Spain have requested that the BWR reactors investigate differences between the methodologies used by the BWRs and PWRs. The developments and improvements done for Cofrentes NPP are detailed. Studies for this plant show that the head loss due to the considered debris is at most half of the limited head loss for the ECCS strainer and the NPSH (Net Positive Suction Head) required for the ECCS pumps is at least three times lower than the NPSH available.

Factores anatómicos, dendrométricos y climáticos implicados en la producción de resina de Pinus pinaster AIT. : aplicación a la mejora de los métodos de resinación

Pinus pinaster Ait. es la conífera con mayor área de distribución en la Península Ibérica y es, a día de hoy, la única especie resinada en nuestro país. La inducción del flujo de resina al exterior para su recolección a través de distintos tipos de heridas ha sido practicada desde hace miles de años por distintas culturas. En todos los casos, las técnicas desarrolladas se basan en la estimulación del característico sistema de defensa de las pináceas. En los últimos siete años se viene observando una tendencia de incremento sustancial de la superficie resinada en España, acompañada por avances tecnológicos dirigidos a la mecanización y mejora de estimulantes. El aprovechamiento resinero se perfila como un sector estratégico en la generación de empleo rural y la conservación de ecosistemas. La industria resinera demanda métodos de extracción más eficaces, una selvicultura adecuada y actualizada, y condiciones laborales de los resineros más dignas con objeto de llegar a ser competitiva en el mercado internacional. Este trabajo se centra en ampliar el conocimiento sobre el sistema de defensa de P. pinaster, concretamente sobre las estructuras y procesos que pueden afectar a la producción de resina. Se analizan las relaciones entre las características anatómicas del xilema, destacando las relacionadas con los canales resiníferos, las variables dendrométricas y dasométricas de la masa y el flujo de resina (objetivo 1). Se estudia cómo estas relaciones son moduladas por las heridas de resinación dependiendo de la técnica de resinación aplicada (objetivo 2), el clima y el balance hídrico del suelo (objetivo 3). El material vegetal, las muestras de suelo y los datos de producción de resina y climáticos usados en esta tesis han sido recogidos en tres montes de utilidad pública; MUP 101 en Armuña, MUP 108 en Melque de Cercos y MUP 117 en Nieva (en esta última solo se recogieron los datos de producciones), todos ellos pinares monoespecíficos de P. pinaster localizados en la denominada Tierra de Pinares Segoviana. En los árboles de nuestro estudio se han aplicado cuatro métodos de resinación: método de pica de corteza con estimulante y método mecanizado con estimulante, ambos en sentido ascendente y descendente. En los trabajos realizados para el análisis de la influencia de la anatomía constitutiva en la producción de resina (objetivo 1) y el efecto del clima (objetivo 3), se obtuvieron muestras del xilema de 26 árboles resinados en Melque de Cercos y Armuña y 12 árboles control sin resinar. Para caracterizar los pies estudiados, se midió la altura, diámetro normal y porcentaje de copa viva. Las muestras de tejido fueron recogidas en una zona del tronco a una distancia del límite de la herida considerada en la bibliografía como no afectada (anatomía constitutiva). Para el análisis de las alteraciones anatómicas inducidas por la herida (objetivo 2), se recogieron muestras en ocho de los individuos en los que se habían realizado los distintos métodos de resinación descritos y en cinco árboles control. Se obtuvieron ocho muestras de tejido distribuidas en la parte superior, inferior, lateral y centro de la herida de cada uno de los árboles resinados. Para establecer las diferencias en la producción de resina según el método de resinación, se analizaron las producciones de 561 árboles resinados en 2012 con estos cuatro métodos en Nieva. Los principales resultados de estos trabajos muestran que la producción de resina está ligada al volumen de canales (axiales y radiales) y a la frecuencia de canales radiales existentes en el árbol antes de efectuar ninguna herida (sistema constitutivo). De esta manera, los árboles grandes productores de resina mostraron una red de canales más densa que aquellos con producciones medias. Una vez realizada la herida de resinación, observamos una disminución del ancho del anillo de crecimiento y del tamaño medio de los canales axiales a la vez que se incrementaba la frecuencia y área ocupada por mm2 de anillo de estos canales. Estos cambios perduraron en el árbol durante al menos tres años y fueron distintos dependiendo de la localización en el entorno de la herida y del método de resinación. Las respuestas más intensas a la herida se observaron el año siguiente a la realización de la misma, en dirección axial, para las distancias más próximas al límite de la herida y para los métodos de resinación en sentido ascendente. Además, se ha constatado que como consecuencia de las heridas de resinación se produjeron cambios en la anatomía del xilema en zonas alejadas de la herida, tanto en el año de la herida como años posteriores. Es decir, se observó una respuesta sistémica del árbol. Respecto al papel del clima como regulador de la respuesta del árbol, se ha evidenciado que la temperatura, la radiación y la ETP influyeron en la producción de resina, no solo durante la campaña de resinación, sino también durante los meses anteriores. El déficit hídrico favoreció la producción y la formación de canales axiales pero, a partir de un determinado umbral, esa relación se invirtió y las lluvias estivales incrementaron la producción. Algunas de estas variables climáticas se asociaron a cambios en el tamaño y frecuencia de las estructuras secretoras, las cuales posiblemente modulan la respuesta defensiva de la planta. La dendrometría del árbol (evaluada a través del diámetro normal, altura y porcentaje de copa viva), la densidad de la masa y el tipo de suelo influyeron en el potencial de producción de resina de P. pinaster. Árboles más vigorosos, parcelas con menores densidades y suelos con más capacidad para la retención de agua y nutrientes presentaron producciones mayores. Estos trabajos se complementan en anexos con una caracterización del sistema socio-ecológico del pinar en resinación. En ese trabajo se identifican sus potenciales servicios ecosistémicos y se evalúa su grado de vinculación con el aprovechamiento resinero con objeto de valorar su funcionalidad y aproximar una valoración económica de modo que sea posible apreciar la importancia económica de los mismos. Para concluir, podemos resaltar que son necesarios más trabajos de carácter científico para avanzar en la comprensión de los procesos anatómicos y fisiológicos que regulan la secreción de resina en P. pinaster y sus interacciones con el medio. Esto permitiría avances certeros hacia el desarrollo de métodos de extracción más eficaces, una selvicultura óptima, el reconocimiento de los beneficios socio-ecológicos y económicos del aprovechamiento y, de manera general, una bibliografía amplia y fiable para la consulta y desarrollo de futuras mejoras que posibiliten la reactivación y conservación de la resinación como aprovechamiento rentable. ABSTRACT Pinus pinaster Ait. is the most widespread conifer in Spain and is now the only species tapped for its oleoresin. External induction of resin secretion, based on the defense system of Pinus trees, has been performed by humans since Classical times through various methods. The socio-economic implication of this practice in Spain justifies a new approach to improve tapping methodology and understand the effects of this activity on the tree. In the last five years, sharp increases in the price of natural resins, accompanied by technological advances directed toward mechanization, have made resin tapping a strategic activity for rural development and forest conservation. The resin industry demands more efficient tapping methods and forest management plans as a way to increase competitiveness in a global market. In this way, this work focuses on the study of the defense system of P. pinaster, with the aim to understand the effects of anatomical and physiological characteristics and environmental conditions on resin yield. The relationships between anatomical variables -with special focus on resin canals-, dendrometric and dasometric variables, and resin yield will be evaluated (objective 1). The tapping wound effects (objective 2) and the intra- and inter-annual variability of climate conditions and soil water availability influence (objective 3) on resin yield will be also studied. The plant and soil material and the resin yield and climatic data used in this thesis have been collected in stands of three public forests of P. pinaster; Armuña, Melque de Cercos and Nieva, located in Segovia (Central Spain). Trees were tapped using two different methods: mechanized or traditional tool, in both upwards and downwards direction. Twenty-six tapped trees of contrasting resin yield classes and twelve non-tapped (control) trees, growing in two locations (Armuña y Melque de Cercos) with the same climate but different stand density and soil characteristics, were selected for studying the role of tree size, xylem anatomy at distal parts aside from the tapping wound (objective 1) and climate influence (objective 3) on resin yield. Concerning the tree defenses induced by the tapping wound (objective 2), the xylem of eight trees, tapped with the two described methods in both upwards and downwards direction, were analyzed. From each tapped tree, eight cores were collected at different locations and varying distances from the tapping wound. In each core, a histological analysis was made. Growth ring width, earlywood and latewood width, and axial canal frequency, area, mean size and location were measured. The effect of the tapping method on resin yield was assessed in 561 P. pinaster tapped trees in a stand in Nieva. In tissues not affected by the tapping wound, the frequency of radial resin canals and the total volume of resin canals were related to resin yield. The frequency of radial canals and the resin yield were strongly related to tree diameter and percentage of live crown. High area of axial resin canals per mm2 was related to high yielding trees, but only in the location with higher plant density and poorer soil quality. In tapped trees, an increase in axial canal frequency and area was found during the three years following the start of tapping activity, suggesting that canal formation is a systemic induced response to wounding. The highest mean annual resin yield was found using the traditional tool in upwards direction, which also induced the highest increase in axial canal frequency and area. The lowest yield was found for mechanized tapping, which showed no differences between the upwards and downwards directions. The strongest induction of systemic induced responses in terms of resin canal frequency and area was detected one year after tapping for upwards tapping. This suggests the involvement of signaling processes that spread mainly upwards, and the importance of adaptive processes as a defense against periodic insect attacks. Intra-annual variation in resin yield was strongly correlated with temperature, solar radiation, potential evapotranspiration and soil water deficit. Inter-annual variation in resin yield and resin canal abundance were correlated with temperature and water deficit in spring, but above a certain threshold of cumulated water deficit in summer rainfall favored resin yield. Under adverse climate scenarios where resource optimization is desirable, a reduced tapping season during the warmest months (June–September) would be advisable, assuming a very small production loss relative to traditional tapping season. Similarly, in years with a rainy summer and/or dry spring, a slightly longer tapping season could be suggested, as resin yield increases after these events. Tree diameter and percentage of live crown, and radial resin canal frequency could be useful criteria for estimating resin yields in P. pinaster. Vigorous trees in lower density stands and growing up in good quality soils will be the most productive. These conclusions could be applied to improve tapping management and breeding programs. These works are complemented with socio-ecological characterization, the identification of the main ecosystem services and an assessment of the possible economic impact derived from the tapping practice. To conclude, more scientific studies are necessary for understanding the anatomical and physiological processes behind resin synthesis and their interactions with the environment. This would afford further progresses towards an extensive and reliable bibliography and improved tapping methods and optimal selvicultural guide lines.

Emulation of an OWC Ocean Energy Plant With PMSG and Irregular Wave Model

Ocean energy is a promising resource for renewable electricity generation that presents many advantages, such as being more predictable than wind energy, but also some disadvantages such as large and slow amplitude variations in the generated power. This paper presents a hardware-in-the-loop prototype that allows the study of the electric power profile generated by a wave power plant based on the oscillating water column (OWC) principle. In particular, it facilitates the development of new solutions to improve the intermittent profile of the power fed into the grid or the test of the OWC behavior when facing a voltage dip. Also, to obtain a more realistic model behavior, statistical models of real waves have been implemented.

Modeling the Real-Time Use of Reserves in the Joint Energy and Reserve Hourly Scheduling of a Pumped Storage Plant

This paper studies the impact that different approaches of modeling the real-time use of the secondary regulation reserves have in the joint energy and reserve hourly scheduling of a price-taker pumped-storage hydropower plant. The unexpected imbalance costs due to the error between the forecasted real-time use of the reserves and the actual value are also studied and evaluated for the different approaches. The proposed methodology is applied to a daily-cycle and closed-loop pumped-storage hydropower plant. Preliminary results show that the deviations in the water volume at the end of the day are important when the percentage of the real-time use of reserves is unknown in advance, and also that the total income in all approaches after correcting these deviations is significantly lower than the maximum theoretical income.

The interspecific mass–density relationship and plant geometry

We present an a priori theoretical framework for the interspecific allometric relationship between stand mass and plant population density. Our model predicts a slope of −\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} \begin{equation*}\frac{1}{3}\end{equation*}\end{document} between the logarithm of stand mass and the logarithm of stand density, thus conflicting with a previously assumed slope of −½. Our model rests on a heuristic separation of resource-limited living mass and structural mass in the plant body. We point out that because of similar resource requirements among plants of different sizes, a nonzero plant mass–density slope is primarily defined by structural mass. Specifically, the slope is a result of (i) the physical size-dependent relationship between stem width and height, (ii) foliage-dependent demands of conductance, and (iii) the cumulative nature of structural mass. The data support our model, both when the potential sampling bias of taxonomic relatedness is accounted for and when it is not. Independent contrasts analyses show that observed relationships among variables are not significantly different from the assumptions made to build the model or from its a priori predictions. We note that the dependence of the plant mass–density slope on the functions of structural mass provides a cause for the difference from the zero slope found in the animal population mass–density relationship; for the most part, animals do not have a comparable cumulative tissue type.

Molecular cloning of a family of xenobiotic-inducible drosophilid cytochrome P450s: Evidence for involvement in host-plant allelochemical resistance

Cytochrome P450s constitute a superfamily of genes encoding mostly microsomal hemoproteins that play a dominant role in the metabolism of a wide variety of both endogenous and foreign compounds. In insects, xenobiotic metabolism (i.e., metabolism of insecticides and toxic natural plant compounds) is known to involve members of the CYP6 family of cytochrome P450s. Use of a 3′ RACE (rapid amplification of cDNA ends) strategy with a degenerate primer based on the conserved cytochrome P450 heme-binding decapeptide loop resulted in the amplification of four cDNA sequences representing another family of cytochrome P450 genes (CYP28) from two species of isoquinoline alkaloid-resistant Drosophila and the cosmopolitan species Drosophila hydei. The CYP28 family forms a monophyletic clade with strong regional homologies to the vertebrate CYP3 family and the insect CYP6 family (both of which are involved in xenobiotic metabolism) and to the insect CYP9 family (of unknown function). Induction of mRNA levels for three of the CYP28 cytochrome P450s by toxic host-plant allelochemicals (up to 11.5-fold) and phenobarbital (up to 49-fold) corroborates previous in vitro metabolism studies and suggests a potentially important role for the CYP28 family in determining patterns of insect–host-plant relationships through xenobiotic detoxification.

Cotransport of water by the Na+/glucose cotransporter

Water is transported across epithelial membranes in the absence of any hydrostatic or osmotic gradients. A prime example is the small intestine, where 10 liters of water are absorbed each day. Although water absorption is secondary to active solute transport, the coupling mechanism between solute and water flow is not understood. We have tested the hypothesis that water transport is directly linked to solute transport by cotransport proteins such as the brush border Na+/glucose cotransporter. The Na+/glucose cotransporter was expressed in Xenopus oocytes, and the changes in cell volume were measured under sugar-transporting and nontransporting conditions. We demonstrate that 260 water molecules are directly coupled to each sugar molecule transported and estimate that in the human intestine this accounts for 5 liters of water absorption per day. Other animal and plant cotransporters such as the Na+/Cl−/γ-aminobutyric acid, Na+/iodide and H+/amino acid transporters are also able to transport water and this suggests that cotransporters play an important role in water homeostasis.

Molecular recognition of pathogen attack occurs inside of plant cells in plant disease resistance specified by the Arabidopsis genes RPS2 and RPM1

The Arabidopsis thaliana disease resistance genes RPS2 and RPM1 belong to a class of plant disease resistance genes that encode proteins that contain an N-terminal tripartite nucleotide binding site (NBS) and a C- terminal tandem array of leucine-rich repeats. RPS2 and RPM1 confer resistance to strains of the bacterial phytopathogen Pseudomonas syringae carrying the avirulence genes avrRpt2 and avrB, respectively. In these gene-for-gene relationships, it has been proposed that pathogen avirulence genes generate specific ligands that are recognized by cognate receptors encoded by the corresponding plant resistance genes. To test this hypothesis, it is crucial to know the site of the potential molecular recognition. Mutational analysis of RPS2 protein and in vitro translation/translocation studies indicated that RPS2 protein is localized in the plant cytoplasm. To determine whether avirulence gene products themselves are the ligands for resistance proteins, we expressed the avrRpt2 and avrB genes directly in plant cells using a novel quantitative transient expression assay, and found that expression of avrRpt2 and avrB elicited a resistance response in plants carrying the corresponding resistance genes. This observation indicates that no bacterial factors other than the avirulence gene products are required for the specific resistance response as long as the avirulence gene products are correctly localized. We propose that molecular recognition of P. syringae in RPS2- and RPM1-specified resistance occurs inside of plant cells.

A host–guest system to study structure–function relationships of membrane fusion peptides

We designed a host–guest fusion peptide system, which is completely soluble in water and has a high affinity for biological and lipid model membranes. The guest sequences are those of the fusion peptides of influenza hemagglutinin, which are solubilized by a highly charged unstructured C-terminal host sequence. These peptides partition to the surface of negatively charged liposomes or erythrocytes and elicit membrane fusion or hemolysis. They undergo a conformational change from random coil to an obliquely inserted (≈33° from the surface) α-helix on binding to model membranes. Partition coefficients for membrane insertion were measured for influenza fusion peptides of increasing lengths (n = 8, 13, 16, and 20). The hydrophobic contribution to the free energy of binding of the 20-residue fusion peptide at pH 5.0 is −7.6 kcal/mol (1 cal = 4.18 J). This energy is sufficient to stabilize a “stalk” intermediate if a typical number of fusion peptides assemble at the site of membrane fusion. The fusion activity of the fusion peptides increases with each increment in length, and this increase strictly correlates with the hydrophobic binding energy and the angle of insertion.

Independent and combined analyses of sequences from all three genomic compartments converge on the root of flowering plant phylogeny

Plant phylogenetic estimates are most likely to be reliable when congruent evidence is obtained independently from the mitochondrial, plastid, and nuclear genomes with all methods of analysis. Here, results are presented from separate and combined genomic analyses of new and previously published data, including six and nine genes (8,911 bp and 12,010 bp, respectively) for different subsets of taxa that suggest Amborella + Nymphaeales (water lilies) are the first-branching angiosperm lineage. Before and after tree-independent noise reduction, most individual genomic compartments and methods of analysis estimated the Amborella + Nymphaeales basal topology with high support. Previous phylogenetic estimates placing Amborella alone as the first extant angiosperm branch may have been misled because of a series of specific problems with paralogy, suboptimal outgroups, long-branch taxa, and method dependence. Ancestral character state reconstructions differ between the two topologies and affect inferences about the features of early angiosperms.

Proline Accumulation in Maize (Zea mays L.) Primary Roots at Low Water Potentials. II. Metabolic Source of Increased Proline Deposition in the Elongation Zone1

The proline (Pro) concentration increases greatly in the growing region of maize (Zea mays L.) primary roots at low water potentials (ψw), largely as a result of an increased net rate of Pro deposition. Labeled glutamate (Glu), ornithine (Orn), or Pro was supplied specifically to the root tip of intact seedlings in solution culture at high and low ψw to assess the relative importance of Pro synthesis, catabolism, utilization, and transport in root-tip Pro deposition. Labeling with [3H]Glu indicated that Pro synthesis from Glu did not increase substantially at low ψw and accounted for only a small fraction of the Pro deposition. Labeling with [14C]Orn showed that Pro synthesis from Orn also could not be a substantial contributor to Pro deposition. Labeling with [3H]Pro indicated that neither Pro catabolism nor utilization in the root tip was decreased at low ψw. Pro catabolism occurred at least as rapidly as Pro synthesis from Glu. There was, however, an increase in Pro uptake at low ψw, which suggests increased Pro transport. Taken together, the data indicate that increased transport of Pro to the root tip serves as the source of low-ψw-induced Pro accumulation. The possible significance of Pro catabolism in sustaining root growth at low ψw is also discussed.