Patterns of leaf surface wetness for montane and subalpine plants

The frequency and duration of water on leaf surfaces have important consequences for plant growth and photosynthetic gas exchange. The objective of the present study was to compare the frequency and duration of leaf wetness under natural field conditions among species and to identify variation in structural features of leaves that may reduce surface wetness. During June–September 1992 in the central Rocky Mountains (USA), natural leaf wetting due to rain and dewfall was observed on 79 of 89 nights in open meadow habitats compared to only 29 of 89 nights in the understorey. Dew formation occurred at relative humidities that were often well below 100% because of radiational heat exchange with cold night skies and low wind speeds (< 0.5 m s^?1). A survey of 50 subalpine/montane species showed that structural characteristics associated with the occurrence and duration of leaf surface wetness differed among species and habitats. Both adaxial and abaxial surfaces accumulated moisture during rain and dewfall events. Leaf surfaces of open-meadow species were less wettable (P= 0.008), and had lower droplet retention (P= 0.015) and more stomata P= 0.017) than adjacent understorey species. Also, leaf trichomes reduced the area of leaf surface covered by moisture. Ecophysiological importance is suggested by the high frequency of leaf wetting events in open microsites, influences on growth and gas exchange, and correspondence between leaf surface wettability and habitat.     

Characterization of the Topography and Wettability of English Weed Leaves and Biomimetic Replicas

The topography and wettability of the underside of English weed （Oxalis pes-caprae） leaves and of their biomimetic replicas are investigated. Polyvinyl siloxane molds were cast from the leaves and then filled with an epoxy pre-polymer to produce replicas. The particular topographical structures of leaves and replicas were evaluated by optical microscopy and Scanning Electron Microscopy （SEM） analysis. The static wettability of leaves and replicas was assessed by contact angle measurements, while the dynamic wettability was characterized by estimating contact angle hysteresis and studying the dynamic behavior of impacting water droplets. A smooth glass slip and its replica were used as control surfaces. The replica moulding method used was able to transfer the characteristic pattern of irregular 100 μm - 200 μm × 60 μm convex papillae interspersed with stomata of the original leaf to the epoxy replicas. The static contact angle of 143°± 3° and the contact angle hysteresis of 2~ indicate that the underside of the English weed leaf is close to superhydrophobic. The lower contact angles （130° ± 4°） and higher hysteresis （31°） observed for the replica when compared with the original leaves were associated to an inaccurate replication of the chemistry and structures of the three-dimensional wax projections covering the plant surface. Also, trichomes in the original leaves could not be accurately reproduced due to their flexibility and fragility. Differences in wetting behavior were also evident from droplet impact experiments, with rebound regimes prevailing in the original leaves and regimes characterized by higher adhesion and larger dissipation predominating in the replicas. Nevertheless, the morphological features of the leaf transferred to the replica were sufficient to promote a clear hydrophobic behavior of the replica when compared with the smooth epoxy reference surface.     

Seasonal changes of leaf surface contamination in beech, oak, and ginkgo in relation to leaf micromorphology and wettability

The leaf surfaces of beech, oak and ginkgo have been investigated with respect to contamination with particles during one growing season. Based on the observation that particles are removed from water-repellent leaves by rain (Lotus effect) the three species were selected because they differ in leaf surface micromorphology and wettability. Leaves of beech are smooth, lacked wax crystals and were ±wettable. Those of ginkgo were rough because their cells were convex and were densely covered by wax crystals, resulting in permanent water repellency. Leaves of oak were covered by waxes and were water repellent when young, but, a few weeks after leaf expansion had ceased the waxes were rapidly eroded. These differences in wettability resulted in different amounts of contamination. Ginkgo collected a very small number of particles during the whole vegetation period. In beech the contamination was significantly higher, but fairly constant, whereas oak leaves accumulated particles with age.     

Visual estimation of leaf water stress inMercurialis perennis L.

Leaf inclination of three upper leaf pairs of a plant was used as a visual parameter for estimation of leaf (plant) water status inM. perennis. Negative correlation was found between leaf angle and leaf water saturation deficit (WSD). Large angles (between 130 and 80–90°) indicated WSD below approx. 12%, narrow angles expressed negative water balance of the plant and indicated usually strong or moderate water stress. The correlation may be expressed by two regression lines differing in slope. Some differences were observed between three leaf pairs investigated: At the same water deficits, leaves of the first pair (from apex) were inclined to a less degree than leaves of the other two leaf pairs. Leaves of the third pair wilted the most rapidly, therefore their WSD were highest at moderate and strong water stress of the plant. The approximation method tested is suitable for judgment of the plant water status inM. ptrennis and may be used in ecological investigations in forest ecosystems.     

Leaf Surface Wetness and Morphological Characteristics of Valeriana Jatamansi Grown Under Open and Shade Habitats

The present study was carried out to investigate the degree of leaf wetness and its capacity to retain water droplets in relation to leaf morphological characteristics of Valeriana jatamansi J. grown under open and shade habitats. Leaves developed in open habitats had less wettability but higher capacity to retain water droplets and more number of stomata than shade leaves. A significant positive correlation of contact angle () were noticed with trichome length, droplet retention and wax content.     

Die Wasserbenetzbarkeit von Blattoberflächen und ihre submikroskopische Wachsstruktur

Summary The wettability of a leaf surface is defined by the contact angle between a water-droplet and the surface of the leaf.Contact angles of 60–80° were measured on easily wettable leaves. These leaves have no wax on the outer cuticular layer.Contact angles of 130–160° were measured on leaves with a low wettability. These leaves have wax on the outer cuticular layer, which shows submicroscopic structures characteristic of the particular plants.A comparison of the wettability and the different structures of the wax showed no true distinctions.The wax does not adhere strongly to the outer cuticular layer and is cast off in the case of old leaves. Therefore the wetability alters with the age of the leaves. If the wax on young leaves is destroyed or removed by outer influences it can be produced again within a few hours.The rebuilt structure of the wax is not always similar to the original one. When the wax was dissolved by organic solvents no new wax formation was observed.Cigarette smoke greatly increases the wettability of leaves of Tropaeolum majus, parochetus communis and Chelidonium majus without causing a visible alteration of the submicroscopic structure. In this case the leaves recover their water-repellency within a few hours, but not if they were treated with Diesel smoke.When the wax structure was destroyed by fungus or Aleurodina no new wax formation was observed.

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Herrn Professor Dr. Walter Rentschler zum 60. Geburtstag     

Periodicity of leaf growth and leaf dry mass changes in the evergreen and deciduous species of Southern Assam,India

The study described patterns of leaf dry mass change, leaf mass per area (LMA), relative growth rate and leaf life span (LL) for 14 evergreen and 7 deciduous species of a tropical forest of Southern Assam, India. Leaf expansion in both the groups was, in general, completed before June (i.e. well before the onset of monsoon rains). Although leaf dry mass during leaf initiation phase was significantly higher (P < 0.01) in evergreen species than in deciduous species, at the time of full leaf expansion, average leaf dry mass relative to the peak leaf dry mass, realised by the evergreen species was lower (66 %) than for deciduous species (76 %). Leaf dry mass increase in both groups continued after leaf full expansion. Evergreen species had a longer leaf dry mass steady phase than deciduous species (2–6 vs 2–3 months). Average LMA of mature leaves for evergreen species (77.43 g m^?2) was significantly greater than that of deciduous species (48.43 g m^?2). LL ranged from 165 days in Gmelina arborea (deciduous) to 509 days in Dipterocarpus turbinatus (evergreen). LMA was correlated positively with LL, indicating that evergreen species with higher leaf construction cost retain leaves for longer period to pay back. The average leaf dry mass loss before leaf shedding was greater (P < 0.01) for deciduous species (30.29 %) than for evergreen species (18.31 %). Although the cost of leaf construction in deciduous species was lower than for evergreen species, they replace leaves at a faster rate. Deciduous species perhaps compensate the cost involved in faster leaf replacement through higher reabsorption of dry mass during senescence, which they remobilise to initiate growth in the following spring when soil resources remain limiting.     

Leaf rosette closure in the alpine rock species Saxifraga paniculata Mill.: significance for survival of drought and heat under high irradiation

The significance of leaf rosette closure for survival of drought and heat under high irradiation on alpine rock sites was investigated in the cushion forming rosette plant, Saxifraga paniculata Mill. With decreasing water content the leaves fold over the rosette centre reducing reversibly the evaporative leaf surface area by 80%. Internal water redistribution driven by an osmotic gradient from older to younger leaves occurs. The oldest leaves dry out to promote the survival of the individual. Leaf temperatures above 45 °C (which match heat tolerance limits 45–57 °C; LT₅₀) co-occurred with low substrate water potentials (less than – 0·5 MPa) on 11·3% of summer days. Shading by leaves can be crucial to surviving high temperatures as it keeps the rosette centre up to 10 °C colder. Mutual shading prevented sustained drought-induced photoinhibition in upper leaf surfaces at relative water contents below 60%. In exposed lower leaf surfaces restoration of photosystem II took several days. Leaf temperatures above 40 °C (21·3% of summer days) induced photoinhibition in situ. Periods with sufficient water supply can be fully utilized as rehydration is fast ( < 12 h) and exposes the upper leaf surfaces that showed only minor photoinhibition. By reversible leaf rosette closure environmental extremes that otherwise could exceed tolerance are efficiently avoided.     

LEAF TEMPERATURES

Ansari , A. Q., and W. E. Loomis . (Iowa State University, Ames.) Leaf temperatures. Amer. Jour. Bot. 46(10): 713–717. Illus. 1959.—Leaf temperatures were measured with a thermocouple and potentiometer. Readings were taken on leaves of varying thickness, under varying environmental and plant conditions, and during alternating heating and cooling cycles in sun and shade. Leaves tended to assume air temperature. Sunshine heated thin leaves 6–10°C. above the air in about 1 min. Very thick leaves were heated 20°C. above air in 20–30 min. Cooling in still air in shade was at the same rate as heating in sunshine, and the product of this rate times leaf mass in g./cm.² was constant for all leaves tested. Wind at 5 m.p.h. lowered leaf temperature in the sun about half way to air temperature. This cooling effect can result in a reduction of transpiration by wind. Transpiration had a minor effect on leaf temperature. Wilted leaves showed nearly the same temperature response as turgid ones. Dried leaves heated less and cooled faster in shade than transpiring leaves. Vaselined leaves were 1–3°C. warmer than transpiring leaves but showed similar heating and cooling curves.     

EFFECT OF LEAF ANGLE AND ORIENTATION ON PHOTOSYNTHESIS AND WATER RELATIONS IN SILPHIUM TEREBINTHINACEUM

Leaf angle and orientation were measured for 217 leaves from two populations of Silphium terebinthinaceum Jacq., a prairie forb with large, unlobed leaves. Seventy-five percent of leaves measured had an angle of deviation from horizontal of more than 60°, and 60% were oriented within 15° of North. Incident Photon Flux Density (PFD), leaf temperature, photosynthesis, stomatal conductance to CO₂, internal CO₂ concentration, transpiration, and water use efficiency (WUE) were measured on 67 pairs of leaves with the axes oriented in either a North-South (N–S) or East–West (E–W) direction. Leaves with axes oriented N–S intercepted higher levels of PFD during morning and afternoon and exhibited higher diurnal rates of photosynthesis and WUE. Leaf temperature was reduced in N–S leaves during midday as compared to E–W leaves, and was lower in vertical leaves than in those in a horizontal position. Therefore, it was concluded that leaf orientation and verticality enhance carbon gain and minimize water loss—characteristics which may have adaptive significance in a hot, stressful prairie environment.     

How Tough are Sclerophylls?

Fracture toughness was estimated for a 'least tough' path inthe leaves of woody species from three sclerophyllous plantcommunities. Most of the species from Mediterranean, tropicalheath forest and lowland tropical rain forest habitats had verytough leaves, with toughness generally 600-1300 J m^-2, whichis two to four times higher than soft-leaved tropical pioneertrees. The toughest leaf (2032 J m^-2), Parishia insignis, camefrom the canopy of the lowland rain forest. Leaves from theshaded understorey of the rain forest did not appear any lesstough than those from the canopy.Copyright 1993, 1999 AcademicPress Leaf fracture toughness, sclerophylly, Mediterranean vegetation, tropical forest     

Variability of stomatal conductance, leaf anatomy, and seasonal leaf wettability of young and adult European beech leaves along a vertical canopy gradient

This study assessed the variation of leaf anatomy, chlorophyll content index (CCI), maximal stomatal conductance (g _s ^max ) and leaf wettability within the canopy of an adult European beech tree (Fagus sylvatica L.) and for beech saplings placed along the vertical gradient in the canopy. At the top canopy level (CL_28m) of the adult beech, CCI and leaf anatomy reflected higher light stress, while g _s ^max increased with height, reflecting the importance of gas exchange in the upper canopy layer. Leaf wettability, measured as drop contact angle, decreased from 85.5°?±?1.6° (summer) to 57.5°?±?2.8° (autumn) at CL_28m of the adult tree. At CL_22m, adult beech leaves seemed to be better optimized for photosynthesis than the CL_28m leaves because of a large leaf thickness with less protective and impregnated substances, and a higher CCI. The beech saplings, in contrast, did not adapt their stomatal characteristics and leaf anatomy according to the same strategy as the adult beech leaves. Consequently, care is needed when scaling up experimental results from seedlings to adult trees.     

Leaf dynamics of seedlings of rain forest species in relation to canopy gaps

Summary Leaf dynamics of eight tropical rain forest species seedlings was studied in three environments: the shaded forest understorey, a small gap of ±50 m², and a large gap of ±500 m². Leaf production rate and leaf loss rate were enhanced in gaps, and a large gap resulted in larger increases than a small gap. For most species net leaf gain rate was larger in gaps, although this rate was not always largest in the large gap. Leaf loss decreased, and leaf survival percentages increased with increasing shade tolerance of species, indicating a slower leaf turnover for more shade tolerant species. Leaf area growth rate was only partly determined by net leaf gain rate. Ontogenetic effects on leaf size were also important, especially in the large gap. Species which possessed leaves with high specific leaf weight (SLW) showed lower leaf loss rates and higher leaf survival percentages than species with low SLW leaves. Leaf life span seemed to be related to leafcost per unit area. The relation of specific patterns in leaf production and leaf loss to the regeneration mode of the species is briefly discussed.     

The Temperatures of Leaves in Assimilation Chambers, and in the Open

The temperatures of apple leaves in assimilation chambers wereup to 17° C above the temperature of the outside air insunlight. Except when in deep shade, enclosed leaves had temperatureshigher than ambient. Leaves in the open in the sun were often2–3° C above ambient, but the greatest differencemeasured was 4.3° C. In shade, leaves in the open were asmuch as 0.8° C below the air temperature. Laboratory experimentswith an incandescent lamp showed that the temperature differencebetween an enclosed leaf and the outside air increased linearlywith increasing light intensity above a certain value. Belowthis value it is believed that changes in leaf permeabilitywere sufficiently large to affect the rate of transpirationand therefore the leaf excess temperature-light intensity relationship.Under field conditions leaves may not be in a steady state;this gives rise to more variable measurements which may indicatea non-linear relation between leaf excess temperature and lightintensity. Methods of cooling leaves in chambers were examined.Impractically high rates of flow of air at the ambient temperatureare necessary to reduce the temperature of enclosed leaves appreciably.Some reduction of the leaf excess temperature can be obtainedby filtering the infra-red from the incident light, or by usinga chamber made of material which transmits far infra-red, thoughcondensation reduces the effectiveness of the latter measure.Leaves exhibit rapid changes in temperature, so the heatingproblem cannot be circumvented by brief enclosure. The mosteffective of the techniques examined is to use a water-cooledchamber, though the temperatures of the leaf and water differby several degrees centigrade in bright light. A simple solutionto the heating problem for field assimilation measurements hasnot been found.     

Variation in leaf nitrogen and phosphorus stoichiometry in the nitrogen-fixing Chinese sea-buckthorn (Hippophae rhamnoides L. subsp. sinensis Rousi) across northern China

Nitrogen (N) and phosphorus (P) concentrations and N:P ratios in terrestrial plants and their patterns of change along environmental gradients are important traits for plant adaptation to changes. We determined the leaf N and P concentrations of Chinese sea-buckthorn (Hippophae rhamnoides L. subsp. sinensis Rousi), a non-legume species with symbiotic N fixation (SNF), at 37 sites across northern China and explored their geographical patterns in relation to climate and soil factors. (1) The mean leaf N, P, and N:P ratio were 36.5, 2.1 mg g^?1, and 17.6, respectively, higher than the mean values of most shrub species in the region. (2) Leaf N was correlated with soil mineral N in cool areas (mean annual temperature MAT <3 °C) but with temperature in warm areas (MAT >3 °C). The high leaf N and divergent leaf N–soil N relationship suggested the importance of SNF in plant N uptake; SNF increases with temperature and is probably the major N source in warm areas. (3) Leaf P was positively related to mean annual precipitation. Leaf N:P ratio was primarily driven by changes in leaf P. The high leaf P reflected the greater requirements of the N-fixing species for P. Our results represent a major advance in understanding the elemental stoichiometry of non-legume N-fixing plants, indicating high P and N requirements and a shift in N source from SNF to soil as temperature declines. This knowledge will help in assessing the habitat suitability for the species and predicting the species dynamics under environmental changes.     

Leaf dynamics and profitability in wild strawberries

Summary Leaf dynamics and carbon gain were evaluated for two species of wild strawberry, Fragaria virginiana and F. vesca. Five populations on sites representing a gradient of successional regrowth near Ithaca, N.Y., U.S.A., were studied for two or three years each. A computer-based model of plant growth and CO₂ exchange combined field studies of leaf biomass dynamics with previously-determined gas exchange rates to estimate carbon balances of leaves and whole plants in different environments.Leaves were produced throughout the growing season, although there was usually a decline in rate of leaf-production in mid-summer. Leaves produced in late spring had the largest area and longest lifespan (except for overwintering leaves produced in the fall). Specific Leaf Weight (SLW) varied little with time of leaf production, but differed greatly among populations; SLW increased with amount of light received in each habitat. The population in the most open habitat had the least seasonal variation in all leaf characters. F. vesca produced lighter, longer-lived leaves than F. virginiana.Simulations showed that age had the largest effect on leaf carbon gain in high-light environments; water stress and temperature had lesser effects. Leaf carbon gain in lowlight environments was relatively unaffected by age and environmental factors other than light. Leaves in high-light environments had the greatest lifetime profit and the greatest ratio of profit to cost. Increasing lifespan by 1/3 increased profit by 80% in low-light leaves and 50% in high-light leaves. Increasing the number of days during which the leaf had the potential to exhibit high photosynthetic rate in response to high light led to little change in profit of low-light leaves while increasing profit of high-light leaves by 49%.     

Effects of litter types, microsite and root diameters on litter decomposition in Pinus sylvestris plantations of northern China

Background and aims

Litter decomposition is a major process in the carbon (C) flow and nutrient cycling of terrestrial ecosystems, but the effects of litter type, microsite, and root diameter on decomposition are poorly understood.

Methods

Litterbags were used to examine the decomposition rate of leaf litter and roots at three soil depths (5, 10 and 20 cm) over a 470-day period in Pinus sylvestris plantations in northern China.

Results

Leaves and the finest roots decomposed more quickly at 5 cm depth and coarser roots (>1-mm) decomposed more quickly at 10 and 20 cm depth. Roots generally decomposed more quickly than leaf litter, except at 5 cm deep; leaves decomposed more quickly than the coarsest roots (>5-mm). Root decomposition was strongly influenced by root diameter. Leaves experienced net nitrogen (N) immobilization and coarse roots (>2-mm) experienced more N release than fine roots. Significant heterogeneity was seen in N release for fine-roots (<2-mm) with N immobilization occurring in smaller (0.5–2-mm) roots and N release in the finest roots (<0.5-mm).

Conclusions

Soil depth of litter placement significantly influenced the relative contribution of the decomposition of leaves and roots of different diameters to carbon and nutrient cycling.     

Differential leaf traits of a neotropical tree Cariniana legalis (Mart.) Kuntze (Lecythidaceae): comparing saplings and emergent trees

Cariniana legalis is an emergent tree that reaches the upper canopy in Brazilian Semideciduous Forest. Spatial contrasts in microclimatic conditions between the upper canopy and understorey in a forest may affect morpho-physiological leaf traits. In order to test the hypothesis that the upper canopy is more stressful to leaves than a gap environment we compared emergent trees of C. legalis, 28–29 m in height to gap saplings, 6–9 m in height, for the following parameters: leaf area, leaf mass area (LMA the dry weight:leaf area ratio), leaf thickness, leaf anatomical parameters, stomata conductance, and chlorophyll a fluorescence. Leaves from emergent trees had smaller leaf areas but greater LMA compared to saplings. Leaf thickness, palisade layer thickness, and stomatal density were higher for emergent trees than for saplings. The opposite pattern was observed for spongy layer thickness and spongy/palisade ratio. Stomatal conductance was also higher for emergent tree leaves than for sapling leaves, but the magnitude of depression on stomatal conductance near midday was more pronounced in emergent trees. The potential quantum yield of photosystem II, as determined by the F _v/F _m ratio was lower for leaves from saplings. The lower values of stomatal conductance, indicating restriction in CO₂ diffusion into the mesophyll can be related to higher photoinhibition observed in the saplings. Leaves from emergent trees and saplings exhibited similar values for apparent electron transport rates and non-photochemical quenching. Our results suggest that changes in leaf traits could be associated to dry conditions at the upper canopy as well as to the ontogenetic transition between sapling/emergent tree life stages.     

Variability in leaf optical properties among 26 species from a broad range of habitats

Leaves from 26 species with growth forms from annual herbs to trees were collected from open, intermediate, and shaded understory habitats in Mississippi and Kansas, USA. Leaf optical properties including reflectance, transmittance, and absorptance in visible and near infrared (NIR) wavelengths were measured along with leaf thickness and specific leaf mass (SLM). These leaf properties and internal light scattering have been reported to vary with light availability in studies that have focused on a limited number of species. Our objective was to determine whether these patterns in leaf optics and light availability were consistent when a greater number of species were evaluated. Leaf thickness and SLM varied by tenfold among species sampled, but within-habitat variance was high. Although there was a strong trend toward thicker leaves in open habitats, only SLM was significantly greater in open vs. understory habitats. In contrast, leaf optical properties were strikingly similar among habitats. Reflectance and reflectance/transmittance in the NIR were used to estimate internal light scattering and there were strong relationships (r1 > 0.65) between these optical properties and leaf thickness. We concluded that leaf thickness, which did not vary consistently among habitats, was the best predictor of NIR reflectance and internal light scattering. However, because carbon allocation to leaves was lower in understory species (low SLM) yet gross optical properties were similar among all habitats, the energy investment by shade leaves required to achieve optical equivalence with sun leaves was lower. Differences in leaf longevity and growth form within a habitat may help explain the lack of consistent patterns in leaf optics as the number of species sampled increases.     

The Effect of Leaf Water Potential, Leaf Temperature and Light Intensity on Leaf Diffusion Resistance and the Transpiration of Leaves of Malus sylvestris

The relationship between leaf resistance to water vapour diffusion and each of the factors leaf water potential, light intensity and leaf temperature was determined for leaves on seedling apple trees (Malus sylvestris Mill. cv. Granny Smith) in the laboratory. Leaf cuticular resistance was also determined and transpiration was measured on attached leaves for a range of conditions. Leaf resistance was shown to be independent of water potential until potential fell below — 19 bars after which leaf resistance increased rapidly. Exposure of leaves to CO₂-free air extended the range for which resistance was independent of water potential to — 30 bars. The light requirement for minimum leaf resistance was 10 to 20 W m^?2 and at light intensities exceeding these, leaf resistance was unaffected by light intensity. Optimum leaf temperature for minimum diffusion resistance was 23 ± 2°C. The rate of change measured in leaf resistance in leaves given a sudden change in leaf temperature increased as the magnitude of the temperature change increased. For a sudden change of 1°C in leaf temperature, diffusion resistance changed at a rate of 0.01 s cm^?1 min^?1 whilst for a 9°C leaf temperature change, diffusion resistance changed at a rate of 0.1 s cm^?1 min^?1. Cuticular resistance of these leaves was 125 s cm^?1 which is very high compared with resistances for open stomata of 1.5 to 4 s cm^?1 and 30 to 35 s cm^?1 for stomata closed in the dark. Transpiration was measured in attached apple leaves enclosed in a leaf chamber and exposed to a range of conditions of leaf temperature and ambient water vapour density. Peak transpiration of approximately 5 × 10^?6 g cm^?2 s^?1 occurred at a vapour density gradient from the leaf to the air of 12 to 14 g m^?3 after which transpiration declined due presumably to increased stomatal resistance. Leaves in CO₂-free air attained a peak transpiration of 11 × 10^?6 g cm^?2 s^?1 due to lower values of leaf resistance in CO₂ free air. Transpiration then declined in these leaves due to development of an internal leaf resistance (of up to 2 s cm^?1). The internal resistance was masked in leaves at normal CO₂ concentrations by the increase in stomatal resistance.