Received: 12 October 2017 Accepted: 24 April 2018

DOI: 10.1111/pce.13327

ORIGINAL ARTICLE

Insight into the physiological role of water absorption via the

leaf surface from a rehydration kinetics perspective
Paula Guzmán‐Delgado1 | J. Mason Earles2 | Maciej A. Zwieniecki1

1
Department of Plant Sciences, University of
California Davis, Davis, CA 95616, USA
2
School of Forestry & Environmental Studies,
Yale University, New Haven, CT 06511, USA
Correspondence
P. Guzmán‐Delgado, Department of Plant
Sciences, One Shields Avenue, University of
California Davis, Davis, CA 95616, USA.
Email: pguzmandelgado@ucdavis.edu
Funding information
Katherine Esau; Emilio González Esparcia
Abstract
Soil water transported via the petiole is a primary rehydration pathway for leaves of
water‐stressed plants. Leaves may also rehydrate by absorbing water via their epider-
mal surfaces. The mechanisms and physiological relevance of this water pathway,
however, remain unclear, as the associated hydraulic properties are unknown. To gain
insight into the foliar water absorption process, we compared rehydration kinetics via
the petiole and surface of Prunus dulcis and Quercus lobata leaves. Petiole rehydration
could be described by a double exponential function suggesting that 2 partly isolated
water pools exist in leaves of both species. Surface rehydration could be described by
a logistic function, suggesting that leaves behave as a single water pool. Whereas full
leaf rehydration via the petiole required approximately 20 min, it took over 150 and
300 min via the surface of P. dulcis and Q. lobata, respectively. Such differences were
attributed to the high resistance imposed by the leaf surface and especially the cuti-
cle. The minimum resistance to surface rehydration was estimated to be 6.6 × 102
(P. dulcis) and 2.6 × 103 MPa·m2·s·g−1 (Q. lobata), which is remarkably higher than esti-
mated for petiole rehydration. These results are discussed in a physiological context.

KEY W ORDS

cuticle, foliar water uptake, hydraulic resistance, leaf surface, rehydration kinetics

1 | I N T RO D U CT I O N at night or during and after precipitation events. Leaves are generally

Diurnal and seasonal weather variation may lead to a temporal mis-
match between evaporative demand and water availability. Such a
mismatch can induce plant water stress that drives down net primary
productivity, eventually requiring a plant to restore its physiological
capacity via rehydration and recovery (Zwieniecki & Secchi, 2015) or
succumb to mortality (McDowell et al., 2008). Consequently, plant tol-
erance to water stress and the potential for tissue rehydration are two
critical traits that underlie a plant's ability to survive temporal disparity
in water demand and supply. Although stress tolerance is a primary
focus of multiple studies, the physiology of recovery has received
much less attention (e.g., Galmés, Flexas, Savé, & Medrano 2007,
Zarrouk et al., 2016, Skelton, Brodribb, McAdam, & Mitchell, 2017).
Recovery from water stress (either mild or severe) depends on a
plant's ability to rehydrate dehydrated tissues when transpirational
demand is lower than the effective water supply, which may occur
the major plant interface for gas exchange, and they are distally
located to the main source of water, that is, the soil. As a result, they
are the most dehydrated plant organ and ultimately control transpira-
tion rates. As leaves are also the primary sites of photosynthesis, their
rehydration is of key importance for plant physiological recovery from
stress. Leaf rehydration may take minutes to several hours, with full
recovery of leaf physiological function taking even a few days
(Blackman, Brodribb, & Jordan, 2009; Secchi & Zwieniecki, 2011).
The broad range of leaf rehydration dynamics among plant species
depends on the prior level of water stress and leaf rheology
(Zwieniecki, Brodribb, & Holbrook, 2007). Such dynamics have been
studied by supplying water to intact leaves via the petiole (e.g., Boyer,
1974; Brodribb & Holbrook, 2003; Scoffoni, Pou, Aasamaa, & Sack,
2008; Trifilò et al., 2014) assuming that rehydration occurs entirely
from soil recharge. However, several species, spanning rainforest to
desert biomes, are known to absorb atmospheric water via their leaf

1886 © 2018 John Wiley & Sons Ltd wileyonlinelibrary.com/journal/pce Plant Cell Environ. 2018;41:1886–1894.
GUZMÁN‐DELGADO ET AL.
surfaces (Burgess & Dawson, 2004; Martin & von Willert, 2000;
Moser et al., 2016; Yates & Hutley, 1995). Thus, it is interesting to
ask if the absorption of water by the leaf surface can significantly
contribute to overall plant water status and, if so, whether it has
biological relevance at the plant and ecosystem levels (Rundel, 1982;
Stone, 1957).
The petiole can be considered a primary pathway for foliar
rehydration. Rates of rehydration via the leaf vascular network (petiole
rehydration) vary substantially among species and are highly influ-
enced by the hydraulic resistance of the soil–leaf path (Sperry, Hacke,
Oren, & Comstock, 2002) and the within‐leaf connections between
vascular to nonvascular tissues (Zwieniecki et al., 2007). Angiosperm
leaf rheological design links the sites of leaf evaporation to the
vascular system and possibly hydraulically isolates the leaf palisade
mesophyll tissue, providing a buffer from sudden changes in water
availability (Nardini, Raimondo, Lo Gullo, & Salleo, 2010, Rockwell
et al. 2014). Such a design would allow the evaporative pathway to
rapidly rehydrate as evaporative demand decreases but may also
expose a leaf's hydraulic system to sudden failure due to embolism
formation. In fact, an apparent evolutionary shift from leaves with less
hydraulically efficient vasculature to a highly efficient system seems to
promote rehydration from soil recharge (Sack & Scoffoni, 2013).
The absorption of water via aerial organs can be traced back to
early land plants such as liverworts, mosses (Rundel, 1982), and ferns
(Schwerbrock & Leuschner, 2017) through more recently derived
angiosperms (Fernández et al., 2014). Similar to petiole rehydration,
water absorbed through the leaf surface can increase plant water
potential (Berry & Smith, 2014; Cassana, Eller, Oliveira, & Dillenburg,
2016), improve photosynthetic capacity (Simonin, Santiago, &
Dawson, 2009; Smith & McClean, 1989), and facilitate survival in
response to drought conditions (Eller, Lima, & Oliveira, 2013).
Although the exact pathway of foliar water absorption is unknown,
liquid water could enter the leaf via the cuticle (Riederer, 2006;
Schönherr, 1976) and stomatal pores (Burkhardt, Basi, Pariyar, &
Hunsche, 2012; Eichert & Goldbach, 2008). Similarly, the
macroproperties of water transport across leaf surfaces are not well
understood, including basic information such as hydraulic resistance,
rehydration kinetic parameters, and the relationship between leaf
water status and surface absorption properties. So far, no methodo-
logical framework exists for analysing the rehydration kinetics of
water absorbed via the surface of intact leaves. Several studies have
analysed the water permeability of leaf surfaces using isolated cuticles
(Schreiber & Schönherr, 2009), which can provide detailed information
on the mechanisms of the surface water absorption process but also
may lead to biased results as compared with absorption across intact
leaves. In addition, the results obtained in such studies are often
interpreted based on the assumption of a “hydrophobic” cuticle, that
is, composed exclusively of lipids or containing minor amounts of
polysaccharides stemming from the cell wall underneath. However,
recent studies provided evidence for the presence of polysaccharides
throughout the entire cuticle, up to the epicuticular waxes, which may
facilitate the transport of water across the leaf surface (Guzmán,
Fernández, García, et al., 2014; Guzmán, Fernández, Graça, et al.,
2014). This may have implications for the hydraulic strategies
underlying plant‐level recovery and survival following water stress.
1887
Here, we compare the rehydration kinetics of water supplied via
the leaf's vasculature (i.e., petiole) and its surface. Leaf rehydration
via the vasculature is a relatively fast process in terms of providing
water to leaf tissues, especially for cells participating in evapotranspi-
ration. For many species, the kinetics of rehydration can be described
by the combination of two capacitors charging simultaneously by two
different resistors, effectively exhibiting two distinct rehydration
kinetics (Zwieniecki et al., 2007). Such dynamics may arise from the
presence of two hydraulically distinct compartments in leaves
characterized by low and high resistance pathways to the water
supply. The compartment characterized by fast rehydration (i.e., low
hydraulic resistance) in angiosperm leaves was proposed to consist
of the epidermis and bundle sheath extension, whereas the compart-
ment characterized by slow rehydration (i.e., high hydraulic resistance)
was proposed to be formed by mesophyll cells (Zwieniecki et al.,
2007). Whether water absorbed through the leaf surface exhibits
rehydration kinetic patterns similar to the petiole has not been
studied, despite its potential relevance for understanding the restora-
tion of leaf function when recovering from water stress. In this study,
we present a framework for quantifying leaf surface hydraulic
resistance, creating a direct link between petiole and surface rehydra-
tion kinetics. Further, we discuss the biological significance of foliar
water absorption based on the observed kinetic properties.
2 | MATERIAL AND METHODS
2.1 | Plant material
Fully developed, undamaged leaves of almond (Prunus dulcis (Mill.) D.
A. Webb) and valley oak (Quercus lobata Née) were used in this study.
Leaves were selected from south‐east facing branches at sun‐exposed
locations within the canopy of trees growing at the University of
California Davis campus (Davis, USA). Branches from six trees of each
species were collected in the early morning, enclosed in dark plastic
bags containing damp paper towels and transferred to the laboratory.
Leaves from each species were then used for three sets of
measurements:
1. Determination of pressure volume curves and turgor loss points
2. Analysis of rehydration kinetics via the epidermal surface
3. Analysis of rehydration kinetics via the petiole
For rehydration kinetics measurements, branches were bench
dried to different water potentials and placed back in the bags to allow
for water potential equilibration among all leaves on the branch. In this
study, we focused on rehydration of turgid leaves. Thus, prior to
starting the rehydration experiments, leaf water potential was
confirmed to be at the selected value of 80–90% of the turgor loss
point for several leaves along the branch. If the water potential value
was lower than this cutoff value, branches were discarded. If the
water potential value was higher, branches were allowed to continue
drying. Six branches per species (one branch per tree) were finally
used for all the rehydration measurements. Six additional branches
were used for pressure–volume curve determination. In all cases,
1888
leaves were collected from the midportion of the shoots. Following a
given measurement, the leaf surface area was determined by analysing
the scanned material with ImageJ software (ImageJ1.45sNational
Institutes of Health, Bethesda, MD, USA). Then, leaves were
oven‐dried at 80 °C for 3 days, and the final dry weight was recorded.
2.2 | Hydraulic measurements
For all hydraulic measurements, we assumed that the equilibrium
chemical potential of water in the leaf is the driving force responsible
for sorption, that is, generating inward flux. The chemical potential is
often defined in terms of water potential (Ψleaf; Boyer, 1974) and used
in the Ohm's analogy flux (Q) equation as Q = −(Ψleaf − ΨH2Osource) / R
(van den Honert, 1948), where R is the quantitative representation of
hydraulic resistance to flow between the water source and
rehydrating cells. This equation permits the estimation of whole sys-
tem hydraulic resistances to flow without making any assumptions
about the specific nature of the hydraulic paths or spatial distribution
of water pools. We used resistance expressed per surface area
resulting in units (MPa·m2·s·g−1; to calculate the molar hydraulic con-
ductance [L] of the leaf surface one can apply a formula L = 18/R
[mol·MPa−1·s−1·m−2]).
2.3 | Determination of pressure–volume curves and
turgor loss points
Pressure–volume curves for each species were determined following
the bench drying technique used in previous studies (Hinckley,
Duhme, Hinckley, & Richter, 1980, Turner, 1988). In short, branches
were cut under water, placed in plastic bags to prevent transpiration,
and allowed to fully rehydrate. Final Ψleaf ranged from −0.010 to
−0.015 MPa (without any evidence of intercellular leaf space flooding,
i.e., change in colour or refractive properties, see Figure S1). Six leaves
(one leaf per branch) were collected, weighed, and placed on a bench
to desiccate at laboratory conditions (~22 °C with relative humidity at
~30%). Each leaf was reweighed, and its water potential was measured
with a pressure chamber at regular time intervals during the drying
period. The procedure was halted when 10% to 15% of the initial leaf
mass was lost. Leaves were oven‐dried to calculate their relative
water content as relative water content = (fresh weight − dry weight) /
(turgid weight−dry weight). The turgor loss point was then estimated
from the pressure–volume curves for each species (Hinckley et al.,
1980, Tyree & Hammel, 1972).
2.4 | Leaf rehydration through the epidermal surface
The kinetics of water absorption via the leaf surface was analysed
using a custom built “fog chamber”. The chamber was made of
3‐cm‐thick closed‐cell extruded polystyrene foam to provide thermal
insulation. An ultrasonic humidifier producing room temperature
submicron fog droplets was connected to the chamber. A small fan
provided continuous air movement in the chamber to ensure homoge-
neous fog distribution. The temperature inside the chamber was main-
tained at ~22 °C.
Prior to fog exposure, the initial weight of the leaves was deter-
mined. Then, the petioles were sealed with vacuum grease (Dow
GUZMÁN‐DELGADO ET AL.
Corning®, Midland, MI, USA) and wax tape (Parafilm M®, Neenah,
WI, USA), and leaves were weighed again. Leaves were individually
suspended in the fog chamber from the petiole such that no contact
between leaves existed. Fog was present inside the chamber prior
to placing the leaves. Leaves were taken out of the chamber after
20–30 min of exposure to fog and then every ~10 min afterwards. Such
an initial delay was necessary to wet the leaf surface. At every time
interval, leaf surfaces were patted dry with paper towels, and leaves
were weighed. Leaf water potential was measured in a humidified pres-
sure chamber. Only one measurement per leaf was made to avoid
potential changes in leaf absorptive properties due to the handling
process (e.g., damage to the cuticle). Measurements continued until
the water potential of the leaves collected from the chamber was close
to zero. Following the measurements, data corresponding to water
potential values higher (less negative) than −0.4 MPa were discarded
to exclude leaves which had potentially underwent flooding of the inter-
cellular airspace, as indicated from macroscopic changes in leaf refrac-
tive properties (some areas usually bordered by veins became darker).
The cumulative amount of water absorbed through the leaf
surface, ΔM, that is, the difference between initial leaf mass (time,
t = 0) and the mass after fog exposure (t = 20, 30, … [min]) was plotted
against time and fitted with a three‐parameter logistic function with
the following form:
mf
ΔM ¼ t50 −t ; (1)
1þe s
where mf is the mass of leaf when absorption stops, t50 is the time at
which 50% of the cumulative mass is reached, and s is inversely pro-
portional to the rate of water uptake. Mass values were normalized
by the leaf surface area (one leaf side). A logistic function was used
as it is phenomenologically realistic, that is, the initial leaf surface is
dry, and it does not absorb water (i.e., the leaf's mass does not initially
change), and by the end, it stops absorbing water as the driving gradi-
ent approaches zero (i.e., the leaf's mass stops changing). A three‐
parameter logistic function was used because there was no apparent
lower asymptote which led to an insignificant parameter estimate
under the assumption of a four‐parameter logistic equation.
The water potential of each leaf was also plotted against time and
fitted with a four‐parameter logistic function:
Ψf −Ψ0
Ψ ¼ Ψ0 þ t50 −t ; (2)
1þe s
where Ψ0 is the initial water potential of the leaf, and Ψf is the water
potential of the leaf when absorption stops. A four‐parameter logistic
function was used because there was clearly a lower asymptote which
led to significant estimates of all four parameters, justifying its use.
Instantaneous water flux into the leaf was determined as the first
derivative of the mass gain logistic function, and the hydraulic
resistance for a given time was calculated from Ohm's equation using
Ψleaf at time t as the driving force for the corresponding flux.
2.5 | Leaf rehydration via the petiole
Leaf petiole rehydration kinetics was quantified using the reverse
Polish guillotine method, as described by Zwieniecki et al. (2007).
GUZMÁN‐DELGADO ET AL. 1889

Briefly, a 20‐cm‐long branch segment was cut, and the cut‐ends were
sealed with vacuum grease and wax tape. The petiole of a leaf located
about halfway along the stem segment was wrapped in adhesive putty
and wax tape. Grease was added to the guillotine–petiole interface to
seal the entire system. The guillotine with the branch segment was
submerged in water to minimize the potential for introduction of
embolism to the petiole during the cut (i.e., the cut was made under
water). The guillotine razor blade was then deployed, and the leaf
was severed from the branch segment and almost instantaneously
connected to a tube linked to a water container placed on an analytical
balance. The leaf was kept under water during measurements to avoid
evaporative losses through the surface. The flow of water to the leaf
was determined at 1‐s intervals for 3 hr. We used the previously
described double‐exponential fitting method to determine rehydration
kinetics and hydraulic parameters of the studied species assuming two
rehydrating compartments (Zwieniecki et al., 2007). Flow curves were
adjusted by subtracting the mass increment at the end of the rehydra-
tion period to exclude water rehydrating intercellular leaf spaces.
Petiole rehydration measurements were not adjusted for potential sur-
face water uptake as >90% of water was absorbed within few minutes,
whereas surface rehydration required multiple hours to reach the same
level of rehydration. The apparent resistance (R) of each compartment
was estimated using the electrical circuit approach as follows:
Ψ0 τ S
R¼ ; (3)
C
where Ψ0 is the initial leaf water potential, τ is the time constant, S is
the leaf surface area (one leaf side), and C is amount of water absorbed
and stored in each leaf compartment. Due to the sample size and to
avoid the assumption of normality, a Wilconson–Mann–Whitney test
was used to compare the model parameters between species (Fay &
Proschan, 2010). Data analysis was performed using R statistical
software.
3 | RESULTS
3.1 | Rehydration kinetics via the surface
Leaves of P. dulcis and Q. lobata absorbed liquid water through their
epidermal surface, as revealed by the increase in leaf mass and water
potential after exposure to fog (Figure 1). Moreover, prolonged fog

FIGURE 1 Water potential (ψ), mass

increment (ΔM), and estimated conductance
(Ksurf) of Prunus dulcis and Quercus lobata
leaves supplied water via their surface over
time. Grey regions show 95% confidence
interval for the mean predicted value (solid
lines) at each point in time based on the
estimated parameters. Dashed lines and
lighter gray regions indicate estimates for
which no measured data exist but which
facilitate comparison between species
1890 GUZMÁN‐DELGADO ET AL.

exposure allowed leaves of both species to fully rehydrate, as derived
from final water potential values. The rehydration kinetics measured
for the leaves of the two species fit well to three‐ and four‐parameter
logistic models both in terms of mass increment and water potential
over time, respectively (Table 1, Figure 1). Based on model estimates,
P. dulcis leaves absorbed ~15.4 gH2O/m2 via their surface after
approximately 165 min of being subjected to fog, and Q. lobata leaves
absorbed up to ~9.0 gH2O/m2 after approximately 320 min (Table 1).
The absorbed amount of water corresponded well with the expected
amount required to rehydrate leaves to water potential values of
approximately −0.4 MPa (the value selected to exclude potential
flooding of intercellular spaces of the rehydrating leaves). For both
of −1.1 (P. dulcis, 46% decrease) and −1.2 MPa (Q. lobata, 36%
decrease). To facilitate comparisons, changes in conductance (inverse
of resistance) over time are shown (Figure 1). It must be noted that
trends towards zero conductance (infinity in the case of resistance)
both at the beginning and the end of the rehydration process depict
unresolved changes in flux and water potential primarily related to
the phenomena occurring during the wetting of leaf surfaces and
when leaf cells are close to full turgor, respectively.
3.2 | Leaf rehydration kinetics via the petiole
A double‐exponential function fits well to the water absorption

species, the amount of water evaporated as estimated from the bench kinetics via the leaf petiole for the two species (Table 3, Figure 2). This

drying technique and that of water absorbed through the surface to suggests that such rehydration kinetics can be described by the

near full rehydration were not significantly different for the same water sum of the rehydration of two different water pools connected to

potential change (see Figure S1). The t95 and t50 values estimated for the petiole via a low‐resistance path (fast rehydration phase) and a

P. dulcis were lower than for Q. lobata, which reflect a faster rehydration high‐resistance path (slow rehydration phase). Despite P. dulcis leaves

and water potential recovery via surface water uptake in P. dulcis leaves. showing significant variability, the water absorbed during the fast and

The maximum flux of surface absorbed water was almost four slow rehydration phase constituted, on average, 50% each, whereas

times higher across P. dulcis than Q. lobata leaves and attained approximately 75% of the water absorbed by Q. lobata was associated

values of 2.06 versus 0.52 mgH2O·m−2·s−1. Maximum water fluxes with the fast rehydration phase. The time constant (i.e., the time at

corresponded in time with water potentials of approximately which 63% of total water absorption occurred) estimated for the fast

−1.5 MPa for both species, which represents a ~21% (P. dulcis) and phase was higher for P. dulcis than Q. lobata (p < .05), as well as the

~28% (Q. lobata) decrease with respect to initial water potential amount of water absorbed and associated with the slow phase

values. The minimum estimated hydraulic resistance values were (p < .01). Leaves of both species were fully hydrated after approxi-

almost four times higher for Q. lobata than P. dulcis (Table 1). Minimum mately 20 min, with P. dulcis absorbing higher amounts of water than

resistances were reached after 100 and 170 min of fog exposure for Q. lobata (15.1 ± 1.2 vs. ~9.7 ± 0.7 gH2O/m2, p < .01, mean ± standard

P. dulcis and Q. lobata, respectively, when leaves had water potentials error). The hydraulic resistances estimated from petiole rehydration
kinetics for the two species are shown in Table 2. Leaves of Q. lobata
revealed a higher resistance associated with the slow rehydration
TABLE 1 Parameter estimates (Est.), standard errors (SE), and p phase than P. dulcis leaves.
values for logistic model relating water potential (Ψ) and mass incre-
ment (ΔM) to time for Prunus dulcis and Quercus lobata supplied water
via the surface 4 | DISCUSSION
Ψ ΔM
Est. SE p value Est. SE p value In this study, we tested and evaluated a method for analysing water
absorption via the leaf surface that allows for estimating basic hydrau-
P. dulcis
lic parameters, including the change in driving force for water absorp-
Min −2.1 0.2 <.001 ‐‐ ‐‐ ‐‐
tion (leaf water potential), water flux across the surface (change in leaf
Max −0.4 0.2 .02 15.4 1.3 <.001
mass in time), and total resistance to water flow over time. We
t50 92.3 8.0 <.001 71.7 7.7 <.001
selected two different species of importance for California agriculture
s 26.6 8.0 .002 31.2 6.4 <.001
and natural ecosystems. The proposed method is simple and can be
t95 ‐‐ ‐‐ ‐‐ 165 ‐‐ ‐‐
performed in any plant physiology lab. The experimental setup allows
Q. lobata
for prolonged exposure of any leaf to continuous deposition of water
Min −2.0 0.2 <.001 ‐‐ ‐‐ ‐‐
on the leaf surface in an amount that is permissible in nature. Future
Max −0.6 0.1 <.001 9.0 0.5 <.001
studies can apply this technique to investigate more specific mecha-
t50 154.5 22.4 <.001 119.7 11.2 <.001
nisms underlying the biophysical and molecular underpinnings of
s 60.0 18.4 <.001 72.1 12.0 <.001
water transport across leaf surfaces.
t95 ‐‐ ‐‐ ‐‐ 320 ‐‐ ‐‐

Note. For the four‐parameter logistic model, parameter estimates for min-
imum water potential (min; MPa), maximum water potential (max; MPa),
time at which 50% water potential is recovered (t50; min), and strength
of water potential recovery (s) are shown. For the three‐parameter logistic
model, parameter estimates for maximum water absorption (max; g/m2),
time at which 50% total water absorption is reached (t50; min), and the
strength of the water uptake rate (s) are shown. The mean and standard
error for the time at which 95% of total water uptake occurred, t95, is also
given for each species in minutes.
4.1 | Leaf rehydration via the surface is a slow, high‐
resistance process
The rehydration experiments revealed that the absorption of surface
deposited water and further transport towards internal leaf tissues is
much slower than the absorption and redistribution of water via
the vascular pathway (petiole). Leaves were fully hydrated after
GUZMÁN‐DELGADO ET AL. 1891

FIGURE 2 Increment in mass (ΔM) of Prunus

dulcis and Quercus lobata leaves over time
when water was supplied via the petiole. Grey
regions show 95% confidence interval for the
mean predicted value at each point in time
based on the estimated parameters

TABLE 2 Resistance to water transport estimated from the rehy- highlights the importance of soil water for plant recovery from water
dration kinetics of Prunus dulcis and Quercus lobata leaves when water stress. For each species, the initial dehydration level of the leaves used
was supplied via the surface and the petiole in vascular and surface rehydration tests was similar and the total
Resistance (MPa·m2·s·g−1) amount of water absorbed. However, the kinetics of absorption were
P. dulcis Q. lobata vastly different. Vascular rehydration could be described by a double‐
Surface ~6.6 × 10 2
~2.6 × 10 3 exponential model in relation to the presence of two pools or com-
Petiole—fast phase 1
(3.1 ± 1.4) × 10 a 8.6 ± 0.9 a partments of water linked to vascular tissue by a low‐ and high‐resis-
Petiole—slow phase (2.6 ± 1.0) × 102 a (6.3 ± 1.4) × 102 b tance pathway (Zwieniecki et al., 2007). In contrast, the rehydration
via the leaf surface could be described by a single logistic function
Note. Minimum values are shown for surface resistance, and average
values are shown for petiole resistance. Data for petiole resistance are and the occurrence of a single high resistance pathway controlling
means ± standard errors. Within rows, values marked with different letters the kinetics of rehydration. These different kinetics reflect the low
are significantly different according to Wilcoxon–Mann–Whitney test resistance to vascular rehydration versus surface rehydration (when
(p < .05).
expressed leaf area and leaf mass basis). As previously described, the
double‐exponential kinetics of vascular rehydration resulted from leaf
TABLE 3 Means and standard errors (SE) for parameter estimates for internal rheology (Zwieniecki et al., 2007). However, this rheology did
double‐exponential rehydration curves for Prunus dulcis (n = 6) and not appear to play such a significant role during leaf rehydration
Quercus lobata (n = 6) leaves supplied water via the petiole through the surface, as it is most likely masked by the high hydraulic
Mean SE p value resistance of the surface. In this regard, the estimated resistance of
P. dulcis the high‐resistance compartment was approximately 2.5 (P. dulcis)
a 10.6 1.9 <.001 and 4 (Q. lobata) times lower than the minimum value of the
1/b 107.3 47.4 <.001 resistances estimated from surface rehydration kinetics.
c 4.5 1.3 <.001 The high resistance to water absorption conferred by the leaf
1/d 539.4 179.7 <.001 surface and especially the cuticle seems to influence the entire
t95 20.8 5.7 ‐‐ rehydration process, leading to kinetics that could be described by a
Q. lobata logistic model. The slower initial rate of water absorption (the first part
a 6.2 0.8 <.001 of the logistic curves), despite the large gradient of water chemical
1/b 149.6 111.0 <.001 energy across the surface, may reflect the partial wetting of the leaf
c 3.3 1.1 <.001 surfaces and/or an imposed delay in cuticle hydration to significantly
1/d 551.4 120.4 <.001 contribute to leaf water gain. Such a delay may be affected by the
t95 21.8 2.9 ‐‐ structure of the cuticle and associated water distribution at relatively
low hydration stages that hinder water mobility across the matrix
Note. All p values for each parameter fit to data from each leaf fall below
the value given in the table. The four parameters, a (g/m2), b (s−1), c (g/ (Domínguez & Heredia 1999; Riederer, 2006). The absorbed water will
m2), and d (s−1), correspond to the following equation: M(t) = a * (1‐ eventually reach inner cell wall regions and the epidermal cell
exp(−b * t)) + c * (1‐exp(−d * t)), where t is time, and M(t) is the mass of protoplasts contributing to the rehydration of the whole epidermis.
water take up through the petiole at time t. The inverse of parameters a
and b which represents time constant values is shown. The mean and stan- Importantly, the epidermis may be a part of the low‐resistance
dard error for the time at which 95% of total water uptake occurred, t95, pathway that links bundle sheath veins to the hydraulic network
are also given for each species in minutes. allowing for fast redistribution of the water across leaf lamina (Buckley
et al., 2011, Zwieniecki et al., 2007). This redistribution could also lead
approximately 2.8 (t95; P. dulcis) and 5.3 hr (t95; Q. lobata) when water to rewetting the cuticle from its inner side causing additional rear-
was supplied via the surface versus 20 min (both species) via the rangements of its chemical constituents (Guzmán‐Delgado, Fernández,
vascular path, respectively. This order of magnitude difference Venturas, Rodríguez‐Calcerrada, & Gil, 2017). This rearrangement may
1892
favour the movement of water across the cuticle leading to a
reduction in surface hydraulic resistance and an increased flux of
water towards the leaf interior as evidenced by the changes in the
uptake rate of water (logistic curves). These hypotheses are in line
with the model of the formation of an aqueous connection
transversing the cuticle with increasing air humidity (Fernández
et al., 2017) or with changes in the hydration of the cuticle porous
structure and enhancement of vapour transport (Shahraeeni & Or,
2012). The later observed increase in hydraulic resistance (and
decreased flux) could be associated with physico‐chemical changes
in the more rehydrated cuticle, a reduction in the driving force, or
the combination of these two factors. Further, it is not entirely
possible to exclude role of stomata in the absorption process.
Although, stomata were assumed to be closed due to dark conditions,
it is possible that water could be absorbed by exposed parts of the
guard cells and contribute to total surface absorption. It is worth
considering that guard cells do not have plasmodesmatal connections
with other epidermal cells, suggesting such a contribution could be
minimal. Once water passed the cuticle, its redistribution across the
leaf might be affected by leaf rheology, the presence of aquaporins,
and by the vein network.
4.2 | Physiological implications
The amount of water absorbed by the surface over several hours of
exposure to relatively dense fog, approximately 10H2O g/m2 for the
species analysed, allowed leaves to regain their water content and
fully rehydrate. However, this amount of absorbed water is small in
the context of typical transpirational demand. As leaves can transpire
as much as 10 mmolH2O·m−2·s−1 (equivalent of 180 mgH2O·m−2·s−1),
the whole night absorption, the maximum absorption rate, observed
for almond (2.16 mgH2O·m−2·s−1) would only sustain ~400 s of midday
transpiration. The situation might be different if exposure to fog is
followed by a period of low evaporative demand suggesting that foliar
water uptake may be important for ecosystems subjected to frequent
precipitation events or during climate specific seasonal weather
patterns (e.g., spring/fall in Mediterranean climate characterized by
presence of fog and rainfall). For example, it has been suggested that
for redwoods, extended periods of fog may affect leaf gas exchange
throughout much of the day (Simonin et al., 2009). Nevertheless,
despite the low potential for impact of the absorbed volume on whole
tree water status, both studied species seemed to have the ability for
surface water absorption, and it is imperative to discuss the physiolog-
ical significance of this process.
As mentioned above, the amount of water absorbed via the sur-
face is relatively small with respect to transpiration. However, this
water is supplied locally throughout the tree, potentially allowing for
the restoration of lost hydraulic pathways due to water stress. For
example, veins constitute 5–20% of total leaf volume, and xylem itself
may constitute ~0.5% of the leaf volume, which for an average angio-
sperm leaf would be an equivalent of 2 mgH2O/m2, an amount much
smaller than the volume absorbed by the leaf during fog exposure. In
such a case, if the leaf is hydraulically disconnected from the plant,
the observed rehydration level could allow for the full restoration of
xylem transport capacity. In addition, even if xylem transport could
GUZMÁN‐DELGADO ET AL.
not be restored, in a hydraulically isolated leaf, the restoration of
water potential opposite to existing water potential gradients in the
plant could allow for increased activity of the phloem and export of
the leaf constituents to other plant organs. This might be important
for plant growth and survival by supplying leaf assimilates to, for
example, reproductive structures or the stem during leaf senescence.
The rehydration level and the collapse of the negative water potential
in all aerial parts of the plant can also allow for localized restoration of
hydraulic capacity across entire plant (Earles et al., 2016). These
potential physiologically significant aspects of water absorption via
the leaf surface could explain the need for plants to maintain this
pathway for water absorption but deserve further investigation.
From an evolutionary perspective, aerial organs of plants seemed
to have developed in successive attempts to protect chlorophyll‐bear-
ing cells from variations in micro‐environmental conditions, while
allowing for vital exchange of gas with the environment. The forma-
tion of a wettable surface that might facilitate water absorption via
the epidermis could constrain gas exchange (Brewer & Smith, 1995).
However, in many plants, especially those growing in foggy conditions,
the leaf surface with stomata has a lower wettability and droplet
adhesion as compared with the upper surface (Feild et al., 1998,
Goldsmith et al., 2017). Many nonvascular plants (e.g., many
bryophytes) almost entirely rely on absorption of water through their
aerial organs but frequently have external capillary conducting struc-
tures that hold only a well‐defined amount of water and leave much
of the surface free for gas exchange (Dilks & Proctor, 1979). The evo-
lution of vascular tissue and the development of a low‐resistance
hydraulic path to supply water to leaves (leaf venation; Zwieniecki &
Boyce, 2014, Pittermann et al., 2015) allowed for evolution of
epidermal structures, such as the cuticle (Niklas, Cobb, & Matas,
2017), stomata (Brodribb & McAdam, 2017), or trichomes (Brewer &
Smith, 1997) that impact the exchange of matter in the solid
(nutrients), liquid, and gas phase. However, more research is needed
to shed light into the significance of leaf surface features in the
context of whole‐leaf and whole‐plant function.
The leaf cuticle is typically associated with providing a fixed barrier
to excessive transpirational water loss (Yeast and Rose 2013) and, thus,
plays the role of hydraulically separating leaf internal tissue from atmo-
sphere. However, recent research on cuticle structure and chemistry
suggests that, at least for some species (e.g., Eucalyptus camaldulensis,
Eucalyptus globulus, Populus × canescens, and Pyrus communis), the
cuticle is formed in the way that maintains the ability to cross‐transport
liquid water (Guzmán, Fernández, García, et al., 2014; Guzmán,
Fernández, Graça, et al., 2014; Fernández, Guzmán‐Delgado, Graça,
Santos, & Gil, 2016). Moreover, the results obtained in this study
suggest the dynamic control of the leaf rehydration process by the
cuticle, where its chemical and structural relationships in terms of
molecular and macromolecular arrangements are likely to play a signifi-
cant role in water transport kinetics. Additional studies need to be
developed to better understand the relationship between leaf surface
structure, chemistry, and water interactions and the importance for
plant physiological performance. This study provides the first standard-
ized method for measuring hydraulic resistance to surface rehydration,
presenting the opportunity to quantitatively link molecular‐scale
studies on the leaf cuticle to whole‐plant physiological function.
GUZMÁN‐DELGADO ET AL.
ACKNOWLEDGMENTS
P. G.‐D. was supported by “Emilio González Esparcia” (Universidad
Politécnica de Madrid, Spain) and “Katherine Esau” (University of Cal-
ifornia Davis, USA) fellowships.
CONF LICT OF INT E RE ST
The authors do not have conflict of interest to declare.
ORCID
Paula Guzmán‐Delgado http://orcid.org/0000-0002-5264-2430
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SUPPORTI NG INFORMATION
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How to cite this article: Guzmán‐Delgado P, Mason Earles J,
Zwieniecki MA. Insight into the physiological role of water
absorption via the leaf surface from a rehydration kinetics per-
spective. Plant Cell Environ. 2018;41:1886–1894. https://doi.
org/10.1111/pce.13327