Hydraulic analysis of water flow through leaves of sugar maple and red oak

Leaves constitute a substantial fraction of the total resistance to water flow through plants. A key question is how hydraulic resistance within the leaf is distributed among petiole, major veins, minor veins, and the pathways downstream of the veins. We partitioned the leaf hydraulic resistance (R(leaf)) for sugar maple (Acer saccharum) and red oak (Quercus rubra) by measuring the resistance to water flow through leaves before and after cutting specific vein orders. Simulations using an electronic circuit analog with resistors arranged in a hierarchical reticulate network justified the partitioning of total R(leaf) into component additive resistances. On average 64% and 74% of the R(leaf) was situated within the leaf xylem for sugar maple and red oak, respectively. Substantial resistance-32% and 49%- was in the minor venation, 18% and 21% in the major venation, and 14% and 4% in the petiole. The large number of parallel paths (i.e. a large transfer surface) for water leaving the minor veins through the bundle sheath and out of the leaf resulted in the pathways outside the venation comprising only 36% and 26% of R(leaf). Changing leaf temperature during measurement of R(leaf) for intact leaves resulted in a temperature response beyond that expected from changes in viscosity. The extra response was not found for leaves with veins cut, indicating that water crosses cell membranes after it leaves the xylem. The large proportion of resistance in the venation can explain why stomata respond to leaf xylem damage and cavitation. The hydraulic importance of the leaf vein system suggests that the diversity of vein system architectures observed in angiosperms may reflect variation in whole-leaf hydraulic capacity.     

Effects of light intensity and duration on leaf hydraulic conductance and distribution of resistance in shoots of silver birch (Betula pendula)

Variation in leaf hydraulic conductance (K(L)) and distribution of resistance in response to light intensity and duration were examined in shoots of silver birch (Betula pendula Roth). K(L) was determined on detached shoots using the evaporative flux method (transpiration was measured with a porometer and water potential drop with a pressure chamber). Although K(L) depended on light duration per se, its dynamics was largely determined by leaf temperature (T(L)). Both upper-crown leaves and branches developed in well-illuminated environment exhibited higher hydraulic efficiency compared with the lower crown, accounting for vertical trends of apparent soil-to-leaf hydraulic conductance in canopy of silver birch revealed in our previous studies. K(L) varied significantly with light intensity, the highest values for both shade and sun foliage were recorded at photosynthetic photon flux density of 330 micromol m(-2) s(-1). Light responses of K(L) were associated evidently with an irradiance-mediated effect on extravascular tissues involving regulation of cell membrane aquaporins. Effects of irradiance on K(L) resulted in changes of Psi(L), bringing about considerable alteration in partitioning of the resistance between leaves and branch (leafless shoot stem): the contribution of leaves to the shoot total resistance decreased from 94% at -1.0 MPa to 75% at -0.2 MPa. Treatment with HgCl2 decreased hydraulic conductance of both leaves and branches, implying that condition of bordered pit membranes or shoot living tissues may be involved in responses of xylem conductance to Hg2+.     

Sequential leaf senescence and correlatively controlled increases in xylem flow resistance

In bean, Phaseolus vulgaris L. (Contender), the directly measured hydraulic resistance of the xylem pathway between roots and primary leaf pulvinal junctions increased rapidly and progressively from 21 to 28 days after planting. These increases in xylem resistance (+390%) were specifically located in the pulvinal junction of the primary leaf. Moreover, they occurred just prior to the onset of primary leaf yellowing. Developmental increases in xylem hydraulic flow resistance and stomatal resistance, as well as subsequent primary leaf yellowing, were completely prevented by detopping the shoots above the primary leaves at 21 days. Thus, the onset of these senescence-associated symptoms was correlatively controlled. In short-term investigations of the mechanisms involved, flow between petiole and cut tip of excised leaves was rapidly reduced by infiltration of 20 picomoles of soluble dextran into the xylem. Moreover, imbibition of approximately 120 picomoles of dextran by excised leaves increased stomatal resistances. A programmed secretion of hormonal concentrations of similar polysaccharides into specific xylem sections in vivo might provide a mechanism for regulating the partitioning of essential xylem supplies between leaves, thus inducing sequential leaf senescence.     

Leaf hydraulic architecture correlates with regeneration irradiance in tropical rainforest trees

The leaf hydraulic conductance (K(leaf)) is a major determinant of plant water transport capacity. Here, we measured K(leaf), and its basis in the resistances of leaf components, for fully illuminated leaves of five tree species that regenerate in deep shade, and five that regenerate in gaps or clearings, in Panamanian lowland tropical rainforest. We also determined coordination with stomatal characters and leaf mass per area. K(leaf) varied 10-fold across species, and was 3-fold higher in sun- than in shade-establishing species. On average, 12% of leaf hydraulic resistance (= 1/K(leaf)) was located in the petiole, 25% in the major veins, 25% in the minor veins, and 39% outside the xylem. Sun-establishing species had a higher proportion of leaf resistance in the xylem. Across species, component resistances correlated linearly with total leaf resistance. K(leaf) correlated tightly with indices of stomatal pore area, indicating a coordination of liquid- and vapor-phase conductances shifted relative to that of temperate woody species. Leaf hydraulic properties are integrally linked in the complex of traits that define differences in water use and carbon economy across habitats and vegetation zones.     

SECONDARY GROWTH IN NEEDLE LEAVES OF PINUS LONGAEVA (BRISTLECONE PINE) AND OTHER CONIFERS: QUANTITATIVE DATA

Needle leaves of Pinus longaeva can be accurately dated and remain alive on branches for 30 or more yrs, making this species ideal to study secondary growth in leaves. Field observations and regression analysis of needle age versus mean needle length both indicate primary (elongation) growth of needles is completed in the first year. Statistical analysis of cell counts for one- to 33-yr-old needles indicate production along the entire length of needles of 1.0-1.7 cell layers per year of secondary phloem, but no secondary xylem. Microscopic measurements and cell counts reveal that with advancing needle age phloem increases radially and transfusion tissue buckles, but the number of endodermal cells and xylem width do not change. Living phloem remains constant in amount (ca. 9 layers) with advancing needle age, indicating yearly replacement of old by new phloem. For comparison to P. longaeva, needle leaves were also studied for ten other conifer taxa with maximum needle longevities ranging from 3 to 19 yrs. In needles of each taxon no secondary xylem is produced, but secondary phloem production occurs throughout the post-elongation lifespan of the needles regardless of maximum needle longevity.     

Vein recovery from embolism occurs under negative pressure in leaves of sunflower (Helianthus annuus)

Leaf veins undergo cavitation at water potentials (Psi(leaf)) commonly experienced by field-growing plants. Theoretically, embolism reversal should not be possible until xylem pressures rise by several kilopascals of atmospheric pressure, but recent evidence suggests that embolized conduits can be refilled even when surrounded by others at substantial tension (novel refilling). The present study reports 'novel refilling' occurring in leaf veins of sunflower (Helianthus annuus L.) while at Psi(leaf) = -0.33 MPa. Sixty per cent loss of vein hydraulic conductance (K(vein)) was recorded at Psi(leaf) < -0.65 MPa, while stem hydraulic conductance (K(stem)) was unaffected even at Psi(leaf) = -1.1 MPa. Loss of K(vein) was accompanied by stomatal closure. Water-stressed plants (Psi(leaf) = -1.1 MPa) were rehydrated overnight to different target water potentials achieved by using PEG at different concentrations as irrigation medium. K(vein) recovered by 50% at Psi(leaf) = -0.47 MPa and vein refilling was complete at Psi(leaf) = -0.33 MPa, i.e. well below the theoretical limit for conduit refilling (-0.05 MPa as calculated for sunflower minor veins). Mercurials supplied to detached leaves had no effect on the refilling process. Upon rehydration, recovery of K(vein) was not paralleled by recovery of whole-plant hydraulic conductance or leaf conductance to water vapour (g(L)), as a likely consequence of hydraulic failure of other components of the water pathway (root system or extravascular leaf compartments) and/or root-to-leaf chemical signalling. This is the first study providing experimental evidence for 'novel refilling' in a herbaceous dicot and highlighting the importance of this process in the leaf.     

Are needles of Pinus pinaster more vulnerable to xylem embolism than branches? New insights from X‐ray computed tomography

Plants can be highly segmented organisms with an independently redundant design of organs. In the context of plant hydraulics, leaves may be less embolism resistant than stems, allowing hydraulic failure to be restricted to distal organs that can be readily replaced. We quantified drought‐induced embolism in needles and stems of Pinus pinaster using high‐resolution computed tomography (HRCT). HRCT observations of needles were compared with the rehydration kinetics method to estimate the contribution of extra‐xylary pathways to declining hydraulic conductance. High‐resolution computed tomography images indicated that the pressure inducing 50% of embolized tracheids was similar between needle and stem xylem (P_{50 needle xylem} = ?3.62 MPa, P_{50 stem xylem} = ?3.88 MPa). Tracheids in both organs showed no difference in torus overlap of bordered pits. However, estimations of the pressure inducing 50% loss of hydraulic conductance at the whole needle level by the rehydration kinetics method were significantly higher (P_{50 needle} = ?1.71 MPa) than P_{50 needle xylem} derived from HRCT. The vulnerability segmentation hypothesis appears to be valid only when considering hydraulic failure at the entire needle level, including extra‐xylary pathways. Our findings suggest that native embolism in needles is limited and highlight the importance of imaging techniques for vulnerability curves.     

Relative rates of delivery of xylem solute to shoot tissues: Possible relationship to sequential leaf senescence

The rates of delivery of regulatory solutes such as cytokinins and mineral ions from the roots to competing shoot tissues can influence rates of metabolism and development. A 15 min pulse of a synthetic xylem mobile and phloem-immobile solute, acid fuchsin, was used to quantify relative rates of solute delivery to competing organs on excised transpiring bean shoots (Phaseolus vulgaris L. cv. Contender) at different stages of development. Stem, flower and fruit tissues received comparatively low rates of solute delivery. The relative rate of solute delivery to newly opened leaves was initially low, but increased during rapid leaf expansion and then declined progressively as the leaves exceeded 70% of their final area. The relative rate of solute delivery to tissues of the basal primary leaves declined progressively from 2 weeks onwards. This decline appeared to be caused by progressive internally regulated increases in both stomatal resistances and hydraulic resistances to xylem flow up to and into the leaf blade. Thus combined abaxial and adaxial stomatal resistance values in the primary leaves (Rs) increased from 3 to ≥ 7 s cm^?1 between 2 and 5 weeks. Similarly, mean values for the connection resistances (Rc) to hydraulic flow into the primary leaves rose from 7 to 13 TPa · s · m^?1 between 2 and 4 weeks. In the same period pathway resistance from stem to primary leaf petioles (Rp), as determined by direct pressure flow assay, increased from 7 to 15 TPa · s · m^?1. Senescence-associated declines in protein and chlorophyll levels in the primary leaves were initiated in parallel with, or after, declines in relative rates of solute delivery. The provision of extra illumination at the basal leaf level between 2 and 5 weeks did not prevent declines in chlorophyll and soluble protein or increases in stomatal resistance. We suggest that internally programmed changes in the hydraulic architecture of the plant progressively divert xylem-transported root supplies of nutrients and cytokinins from basal to more apical leaves and thus regulate the progressive senescence of leaves from the base upwards.     

Height-related trends in leaf xylem anatomy and shoot hydraulic characteristics in a tall conifer: safety versus efficiency in water transport

Hydraulic vulnerability of Douglas-fir (Pseudotsuga menziesii) branchlets decreases with height, allowing shoots at greater height to maintain hydraulic conductance (K shoot) at more negative leaf water potentials (Psi l). To determine the basis for this trend shoot hydraulic and tracheid anatomical properties of foliage from the tops of Douglas-fir trees were analysed along a height gradient from 5 to 55 m. Values of Psi l at which K shoot was substantially reduced, declined with height by 0.012 Mpa m(-1). Maximum K shoot was reduced by 0.082 mmol m(-2) MPa(-1) s(-1) for every 1 m increase in height. Total tracheid lumen area per needle cross-section, hydraulic mean diameter of leaf tracheid lumens, total number of tracheids per needle cross-section and leaf tracheid length decreased with height by 18.4 microm(2) m(-1), 0.029 microm m(-1), 0.42 m(-1) and 5.3 microm m(-1), respectively. Tracheid thickness-to-span ratio (tw/b)2 increased with height by 1.04 x 10(-3) m(-1) and pit number per tracheid decreased with height by 0.07 m(-1). Leaf anatomical adjustments that enhanced the ability to cope with vertical gradients of increasing xylem tension were attained at the expense of reduced water transport capacity and efficiency, possibly contributing to height-related decline in growth of Douglas fir.     

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.     

Hydraulic efficiency and coordination with xylem resistance to cavitation, leaf function, and growth performance among eight unrelated Populus deltoidesxPopulus nigra hybrids

Tests were carried out to determine whether variations in the hydraulic architecture of eight Populus deltoides×Populus nigra genotypes could be related to variations in leaf function and growth performance. Measurements were performed in a coppice plantation on 1-year-old shoots under optimal irrigation. Hydraulic architecture was characterized through estimates of hydraulic efficiency (the ratio of conducting sapwood area to leaf area, A(X):A(L); leaf- and xylem-specific hydraulic conductance of defoliated shoots, k(SL) and k(SS), respectively; apparent whole-plant leaf-specific hydraulic conductance, k(plant)) and xylem safety (water potential inducing 50% loss in hydraulic conductance). The eight genotypes spanned a significant range of k(SL) from 2.63 kg s(-1) m(-2) MPa(-1) to 4.18 kg s(-1) m(-2) MPa(-1), variations being mostly driven by k(SS) rather than A(X):A(L). There was a strong trade-off between hydraulic efficiency and xylem safety. Values of k(SL) correlated positively with k(plant), indicating that high-pressure flowmeter (HPFM) measurements of stem hydraulic efficiency accurately reflected whole-plant water transport efficiency of field-grown plants at maximum transpiration rate. No clear relationship could be found between hydraulic efficiency and either net CO(2) assimilation rates, water-use efficiency estimates (intrinsic water-use efficiency and carbon isotope discrimination against (13)C), or stomatal characteristics (stomatal density and stomatal pore area index). Estimates of hydraulic efficiency were negatively associated with relative growth rate. This unusual pattern, combined with the trade-off observed between hydraulic efficiency and xylem safety, provides the rationale for the positive link already reported between relative growth rate and xylem safety among the same eight P. deltoides×P. nigra genotypes.     

Changes in leaf hydraulic conductance correlate with leaf vein embolism in<Emphasis Type="Italic"> Cercis siliquastrum</Emphasis> L.

The impact of xylem cavitation and embolism on leaf (K _leaf) and stem (K _stem) hydraulic conductance was measured in current-year shoots of Cercis siliquastrum L. (Judas tree) using the vacuum chamber technique. K _stem decreased at leaf water potentials (Ψ_L) lower than ?1.0 MPa, while K _leaf started to decrease only at Ψ_L L K _leaf changes. Field measurements of leaf conductance to water vapour (g _L) and Ψ_L showed that stomata closed when Ψ_L decreased below the Ψ_L threshold inducing loss of hydraulic conductance in the leaf. The partitioning of hydraulic resistances within shoots and leaves was measured using the high-pressure flow meter method. The ratio of leaf to shoot hydraulic resistance was about 0.8, suggesting that stem cavitation had a limited impact on whole shoot hydraulic conductance. We suggest that stomatal aperture may be regulated by the cavitation-induced reduction of hydraulic conductance of the soil-to-leaf water pathway which, in turn, strongly depends on the hydraulic architecture of the plant and, in particular, on leaf hydraulics.     

Water stress deforms tracheids peripheral to the leaf vein of a tropical conifer

Just as a soggy paper straw is prone to yielding under the applied suction of a thirsty drinker, the xylem tracheids in leaves seem prone to collapse as water potential declines, impeding their function. Here we describe the collapse, under tension, of lignified cells peripheral to the leaf vein of a broad-leaved rainforest conifer, Podocarpus grayi de Laub. Leaves of Podocarpus are characterized by an array of cylindrical tracheids aligned perpendicular to the leaf vein, apparently involved in the distribution of water radially through the mesophyll. During leaf desiccation the majority of these tracheids collapsed from circular to flat over the water potential range -1.5 to -2.8 MPa. An increase in the percentage of tracheids collapsed during imposed water stress was mirrored by declining leaf hydraulic conductivity (K(leaf)), implying a direct effect on water transport efficiency. Stomata responded to water stress by closing at -2.0 MPa when 45% of cells were collapsed and K(leaf) had declined by 25%. This was still substantially before the initial indications of cavitation-induced loss of hydraulic conductance in the leaf vein, at -3 MPa. Plants droughted until 49% of tracheids had collapsed were found to fully recover tracheid shape and leaf function 1 week after rewatering. A simple mechanical model of tracheid collapse, derived from the theoretical buckling pressure for pipes, accurately predicted the collapse dynamics observed in P. grayi, substantiating estimates of cell wall elasticity and measured leaf water potential. The possible adaptive advantages of collapsible vascular tissue are discussed.     

Evidence for xylem embolism as a primary factor in dehydration-induced declines in leaf hydraulic conductance

Hydraulic conductance of leaves (K(leaf)) typically decreases with increasing water stress and recent studies have proposed different mechanisms responsible for decreasing K(leaf) . We measured K(leaf) concurrently with ultrasonic acoustic emissions (UAEs) in dehydrating leaves of several species to determine whether declining K(leaf) was associated with xylem embolism. In addition, we performed experiments in which the surface tension of water in the leaf xylem was reduced by using a surfactant solution. Finally, we compared the hydraulic vulnerability of entire leaves with the leaf lamina in three species. Leaf hydraulic vulnerability based on rehydration kinetics and UAE was very similar, except in Quercus garryana. However, water potentials corresponding to the initial decline in K(leaf) and the onset of UAE in Q. garryana were similar. In all species tested, reducing the surface tension of water caused K(leaf) to decline at less negative water potentials compared with leaves supplied with water. Microscopy revealed that as the fraction of embolized xylem increased, K(leaf) declined sharply in Q. garryana. Measurements on leaf discs revealed that reductions in lamina hydraulic conductance with dehydration were not as great as those observed in intact leaves, suggesting that embolism was the primary mechanism for reductions in K(leaf) during dehydration.     

Hydraulic properties of rice and the response of gas exchange to water stress

We investigated the role of xylem cavitation, plant hydraulic conductance, and root pressure in the response of rice (Oryza sativa) gas exchange to water stress. In the field (Philippines), the percentage loss of xylem conductivity (PLC) from cavitation exceeded 60% in leaves even in watered controls. The PLC versus leaf water potential relationship indicated diurnal refilling of cavitated xylem. The leaf water potential causing 50 PLC (P(50)) was -1.6 MPa and did not differ between upland versus lowland rice varieties. Greenhouse-grown varieties (Utah) were more resistant to cavitation with a 50 PLC of -1.9 MPa but also showed no difference between varieties. Six-day droughts caused concomitant reductions in leaf-specific photosynthetic rate, leaf diffusive conductance, and soil-leaf hydraulic conductance that were associated with cavitation-inducing water potentials and the disappearance of nightly root pressure. The return of root pressure after drought was associated with the complete recovery of leaf diffusive conductance, leaf-specific photosynthetic rate, and soil-leaf hydraulic conductance. Root pressure after the 6-d drought (61.2 +/- 8.8 kPa) was stimulated 7-fold compared with well-watered plants before drought (8.5 +/- 3.8 kPa). The results indicate: (a) that xylem cavitation plays a major role in the reduction of plant hydraulic conductance during drought, and (b) that rice can readily reverse cavitation, possibly aided by nocturnal root pressure.     

Winter at the alpine timberline. Why does embolism occur in norway spruce but not in stone pine?

Conifers growing at the alpine timberline are exposed to frost drought and freeze-thaw cycles during winter-stress factors known to induce embolism in tree xylem. The two dominant species of the European Central Alps timberline were studied: Norway spruce (Picea abies [L.] Karst) and stone pine (Pinus cembra), which usually reaches higher altitudes. We hypothesized to find embolism only at the timberline and to observe less embolism in stone pine than in Norway spruce due to avoidance mechanisms. Seasonal courses of embolism and water potential were studied at 1,700 and 2,100 m during two winter seasons and correlated to vulnerability (to drought-induced embolism), leaf conductance, and micrometeorological data. Embolism was observed only at the timberline and only in Norway spruce (up to 49.2% loss of conductivity). Conductivity losses corresponded to low water potentials (down to -3.5 MPa) but also to the number of freeze-thaw events indicating both stress factors to contribute to embolism induction. Decreasing embolism rates-probably due to refilling-were observed already in winter. Stone pine did not exhibit an adapted vulnerability (50% loss of conductivity at -3.5 MPa) but avoided critical potentials (minimum -2.3 MPa): Cuticulare conductance was 3.5-fold lower than in Norway spruce, and angles between needles and axes were found to decrease in dehydrating branches. The extent of conductivity losses in Norway spruce and the spectrum of avoidance and recovery mechanisms in both species indicates winter embolism to be relevant for tree line formation.     

Hydraulic design of pine needles: one-dimensional optimization for single-vein leaves

Single-vein leaves have the simplest hydraulic design possible, yet even this linear water delivery system can be modulated to improve physiological performance. We determined the optimal distribution of transport capacity that minimizes pressure drop per given investment in xylem permeability along the needle for a given length without a change in total water delivery, or maximizes needle length for a given pressure difference between petiole and needle tip. This theory was tested by comparative analysis of the hydraulic design of three pine species that differ in the length of their needles [Pinus palustris (Engl.) Miller, approximately 50 cm; Pinus ponderosa Lawson & Lawson, approximately 20 cm and Pinus rigida Miller, approximately 5 cm]. In all three species, the distribution of hydraulic permeability was similar to that predicted by the optimum solution. The needles of P. palustris showed an almost perfect match between predicted and actual hydraulic optimum solution, providing evidence that vein design is a significant factor in the hydraulic design of pine leaves.     

Water balance in Hemizonia luzulifolia: the role of extracellular polysaccharides

Abstract. The role of extracellular water in ameliorating drought stress was examined in Hemizonia luzulifola , an annual composite that comprises two subspecies differing significantly in the amount of extracellular polysaccharide within basal leaves. Rosette leaves of the high polysaccharide (HP) ssp were more than 30% pectin on a dry weight basis in contrast to only 4% in the low polysaccharide (LP) ssp. Concomitant with this difference in polysaccharide content was a significant difference in the dehydration response of leaves. Near full hydration, relative capacitances of HP leaves (0.7 MPa^-1) were an order of magnitude greater than the LP leaves (0.08 MPa^-1). Relative capacitance of the polysaccharide alone was 1.5 MPa^-1. The weight of water per unit leaf area was not significantly different in the two taxa. Nevertheless, the pectin-like polysaccharides significantly altered the distribution of metabolically available water from primarily cell-stored water in the LP ssp to apoplasmic and symplasmic capacitors in leaves of the HP ssp. The consequence of this apoplasmic water store with colloidal properties is that the Ψ-dependence of leaf capacitance ceases to be linked directly to cell water relations. Transfer resistances for water movement between capacitors and the xylem near full hydration (0 to -0.5 MPa) were significantly larger in the HP leaf. This difference in transfer resistance was interpreted to be the result of a large resistance to water movement between the polysaccharide and the xylem. Because of these large transfer resistances, the apoplasmic capacitor probably buffers cells at lower water potentials under transient water fluxes than expected from laboratory measurements made during slow desiccation. Field measurements support this conclusions; HP leaves were better buffered than LP leaves at midday water potentials.     

Heterologous array analysis in Pinaceae: hybridization of Pinus taeda cDNA arrays with cDNA from needles and embryogenic cultures of P. Taeda, P. Sylvestris or Picea abies

Hybridization of labelled cDNA from various cell types with high-density arrays of expressed sequence tags is a powerful technique for investigating gene expression. Few conifer cDNA libraries have been sequenced. Because of the high level of sequence conservation between Pinus and Picea we have investigated the use of arrays from one genus for studies of gene expression in the other. The partial cDNAs from 384 identifiable genes expressed in differentiating xylem of Pinus taeda were printed on nylon membranes in randomized replicates. These were hybridized with labelled cDNA from needles or embryogenic cultures of Pinus taeda, P. sylvestris and Picea abies, and with labelled cDNA from leaves of Nicotiana tabacum. The Spearman correlation of gene expression for pairs of conifer species was high for needles (r(2) = 0.78 - 0.86), and somewhat lower for embryogenic cultures (r(2) = 0.68 - 0.83). The correlation of gene expression for tobacco leaves and needles of each of the three conifer species was lower but sufficiently high (r(2) = 0.52 - 0.63) to suggest that many partial gene sequences are conserved in angiosperms and gymnosperms. Heterologous probing was further used to identify tissue-specific gene expression over species boundaries. To evaluate the significance of differences in gene expression, conventional parametric tests were compared with permutation tests after four methods of normalization. Permutation tests after Z-normalization provide the highest degree of discrimination but may enhance the probability of type I errors. It is concluded that arrays of cDNA from loblolly pine are useful for studies of gene expression in other pines or spruces.     

Stem and leaf hydraulic properties are finely coordinated in three tropical rain forest tree species

Coordination of stem and leaf hydraulic traits allows terrestrial plants to maintain safe water status under limited water supply. Tropical rain forests, one of the world's most productive biomes, are vulnerable to drought and potentially threatened by increased aridity due to global climate change. However, the relationship of stem and leaf traits within the plant hydraulic continuum remains understudied, particularly in tropical species. We studied within‐plant hydraulic coordination between stems and leaves in three tropical lowland rain forest tree species by analyses of hydraulic vulnerability [hydraulic methods and ultrasonic emission (UE) analysis], pressure‐volume relations and in situ pre‐dawn and midday water potentials (Ψ). We found finely coordinated stem and leaf hydraulic features, with a strategy of sacrificing leaves in favour of stems. Fifty percent of hydraulic conductivity (P₅₀) was lost at ?2.1 to ?3.1 MPa in stems and at ?1.7 to ?2.2 MPa in leaves. UE analysis corresponded to hydraulic measurements. Safety margins (leaf P₅₀ – stem P₅₀) were very narrow at ?0.4 to ?1.4 MPa. Pressure‐volume analysis and in situ Ψ indicated safe water status in stems but risk of hydraulic failure in leaves. Our study shows that stem and leaf hydraulics were finely tuned to avoid embolism formation in the xylem.