Tree Physiology 29, 375–388
doi:10.1093/treephys/tpn032
Seasonal effects of deficit irrigation on leaf photosynthetic traits
of fruiting and non-fruiting shoots in almond trees
PEDRO A. NORTES,1 MARIA M. GONZALEZ-REAL,1,2 GREGORIO EGEA1
and ALAIN BAILLE1
1
Universidad Politécnica de Cartagena, Escuela Te´cnica Superior de Ingenieros Agrónomos, Área de Ingenierı´a Agroforestal,
Paseo Alfonso XIII, 48, 30203, Cartagena, Spain
2
Corresponding author (mayla.gonreal@upct.es)
Received July 17, 2008; accepted October 27, 2008; published online January 13, 2009
Summary We investigated seasonal trends in, and rela-
tionships between, leaf structural properties, leaf nitrogen
concentration, and maximum (Am) and potential (Ap) leaf
net CO2 assimilation of 1-year-old fruiting (f) and current-
year non-fruiting (nf) shoots in 5-year-old almond trees
(Prunus dulcis (Mill.) D.A. Webb cv Marta). These trees
had been subjected in the previous 4 years to either full
irrigation (FI regime) or sustained deficit irrigation (DI) at
50% of standard crop evapotranspiration during the entire
growing season (DI regime) in the semiarid climate of
southeast Spain. Measurements were made during an entire
growing season on sun-exposed leaves. Leaf dry mass per
unit area (Wa), area and dry-mass-based leaf N concentra-
tions (Na and Nw, respectively), and area and dry-mass-
based Am (Ama and Amw, respectively) were lower in f-leaves
than in nf-leaves. Changes in leaf structural attributes
induced by DI were more pronounced in nf-leaves than in
f-leaves, the latter being little affected. Over the entire
growth season, Am and Ap were correlated negatively with
Wa and positively with Nw for both the leaf classes and the
irrigation regimes. When calculated with respect to total
leaf N concentration, maximum photosynthetic nitrogen-
use efficiency (PNUEm) was significantly higher in f-leaves
than in nf-leaves, with no significant differences between the
leaf classes among the irrigation regimes. However, when
PNUEm was calculated with respect to photosynthetic N,
no significant effect of leaf class or irrigation regime was
observed. Overall, our results showed that DI and FI trees
exhibited similar seasonal patterns of leaf structural
properties and maximum and potential leaf net CO2
assimilation rates, but there were distinct N-allocation
patterns between f- and nf-leaves. In the DI treatment, leaf
structural adjustments appeared to operate to maintain a
high N status in the leaves of fruit-bearing shoots, to the
detriment of N resources allocated to vegetative shoots.
Keywords: leaf N concentration, phenological stage, pho-
tosynthetic capacity, photosynthetic nitrogen-use efficiency,
Prunus dulcis, specific leaf area.
 The Author 2009. Published by Oxford University Press. All rights reserved.
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Introduction
Deficit irrigation (DI) strategies can regularize or increase,
or both, almond productivity when a limited amount of
irrigation water is available (Goldhamer et al. 2006). These
strategies, which include regulated DI (Chalmers et al.
1981), sustained DI (Girona et al. 2005, Goldhamer et al.
2006), and partial root-zone drying (Dry and Loveys
1999, Kang et al. 2003), can substantially reduce tree water
consumption without causing a significant negative effect
on tree productivity (Girona et al. 1997, Goldhamer and
Viveros 2000, Esparza et al. 2001a, 2001b, Romero et al.
2004, Girona et al. 2005). One of the prerequisites for val-
uation and further implementation of best DI management
practices is a detailed knowledge of plant physiologic
responses to abiotic stresses, while accounting for leaf
age, plant phenology (Dungan et al. 2003, Xu and Baldoc-
chi 2003, Muroaka and Koizumi 2005) and fruit load
(DeJong 1986, Palmer et al. 1997, Syvertsen et al. 2003).
Studies on the application of DI to the almond tree have
provided valuable information, especially on leaf net CO2
assimilation (DeJong 1983, Wartinger et al. 1990, Higgins
et al. 1992, Matos et al. 1998, Romero et al. 2004, Rouhi
et al. 2007) and an influence of natural or imposed
drought-induced stress on photosynthetic attributes (Klein
et al. 2001, Heilmeier et al. 2002, Romero and Botı́a
2006). Details related to nutrient deficiency (Basile et al.
2003), species, cultivar type and rootstocks (Matos et al.
1997, De Herralde et al. 2003, Rouhi et al. 2007) and leaf
ontogeny (Wartinger et al. 1990, Matos et al. 1998, Klein
et al. 2001, Romero and Botı́a 2006) are also available.
Knowledge about how water stress can modify the
functioning of different types of shoots in the almond tree
is needed, because it is likely to affect almond yield
determinants. Studies on fruit trees have shown differences
in leaf structural and physiologic traits between shoot types
(Fujii and Kennedy 1985, Syvertsen et al. 2003, Zhang et al.
2005), associated with the presence or absence of fruits on
shoots. Recently, Heerema et al. (2008) reported that the
376 NORTES, GONZALEZ-REAL, EGEA AND BAILLE
survival of almond rosette-like short shoots (spurs) over
winter was strongly related to their leaf area per unit of leaf
mass. Although a clear seasonal trend in leaf traits, which
can vary considerably over time with leaf age, has been
observed (Wilson et al. 2000), it has not been clearly estab-
lished if leaf age dominates other factors such as soil water
availability in the almond tree.
To our knowledge, the long-term effects of DI strategies
on the response of leaf structural and physiologic traits of
fruit-bearing shoots and vegetative shoots, and on the pat-
tern of N allocation to these classes of foliage, have not
been documented. Such responses might induce changes
in the pool of carbohydrate reserves, which are known to
affect the renewal of fruiting position, as well as the amount
of reserves available at the onset of a new growth cycle,
leading to subsequent yield reduction in the following year
(Esparza et al. 2001a, 2001b). In early foliating species,
growing shoots and developing fruits are strong sinks for
assimilates, which compete for the reserves stored in woody
plant parts during the previous growing season (Weinbaum
et al. 1987, Millard 1996, Nii 1997, Rosecrance et al. 1998,
Youssefi et al. 2000, Bi et al. 2003). The influence of fruits
(sinks) in attracting assimilates from the proximal leaves
(Nii 1997, Syvertsen et al. 2003) and competition for N
resources between leaves and fruits (Youssefi et al. 2000)
can lead to lower N concentration in leaves near the fruit,
as observed in citrus trees (Syvertsen et al. 2003). In walnut
trees, a net decrease in leaf N concentration as a result of
protein degradation (resorption) can occur in response to
the demands of developing fruits (Weinbaum et al. 1994),
but this pattern has not clearly been demonstrated in
almond trees (Weinbaum and Muraoka 1986). Stress con-
ditions have been reported to affect leaf N concentration
(Reich et al. 1989). Because leaves are an important source
of N for tree reserves, restricting the water supply by DI
might have a negative effect on N accumulation in woody
structures (Bi et al. 2003). It has been observed that, after
harvest, N storage in perennial tree parts depends much
more on N recycling from leaves than on soil N uptake
(Rosecrance et al. 1998, Esparza et al. 2001a), and this stor-
age can provide up to 50% of the N used over the growing
season (Weinbaum et al. 1987).
Romero et al. (2004) reported a significant reduction in
the photosynthetic capacity of young branches of almond
trees under pre-harvest conditions of mild to moderate soil
water deprivation, whereas the accumulation of dry matter
in the kernel was little affected compared with the fully irri-
gated trees. Because shoots appear to function as autono-
mous structures with respect to assimilation and use of
carbon in many tree species (Sprugel et al. 1991), the sus-
tained sink activity of fruits under conditions of water stress
might indicate that the almond tree tends to preserve the
photosynthetic capacity of fruit-bearing shoots to the detri-
ment of the vegetative shoots. This suggestion is supported
by the observed differences in nonstructural carbohydrate
TREE PHYSIOLOGY VOLUME 29, 2009
concentration between shoot types of dormant almond
trees that had previously been subjected to water stress dur-
ing the harvest period (Esparza et al. 2001a).
The results of the cited studies suggest that a DI strategy
applied for several consecutive years could lead to signifi-
cant changes in the morphologic and physiologic traits of
almond leaves. We hypothesized that vegetative and fruit-
bearing shoots exhibit a differential response to water stress
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through changes in leaf structural and physiologic traits. To
test this hypothesis, we characterized the seasonal trend in
leaf traits, and investigated whether they vary (1) with leaf
age and class (fruiting versus non-fruiting shoots) and (2)
between irrigation regimes (full irrigation (FI) versus sus-
tained DI), and investigated to what extent the relationships
between leaf traits are conserved or co-vary across leaf clas-
ses and irrigation regimes. We also examined the effect of
irrigation treatment on the photosynthetic nitrogen-use effi-
ciency (PNUE) of the two leaf classes.
Materials and methods
Site description
The study was conducted during the 2004 growing season in a
1-ha plot of 5-year-old almond (Prunus dulcis (Mill.) D.A.
Webb cv Marta) trees that had been grafted on ‘Mayor’ root-
stock and planted in December 1999 at a spacing of
6 m · 7 m in an almond orchard at the Agricultural Exper-
imental Station of the University of Cartagena (3735 0 N and
059 0 W). The climate at the site is mediterranean with low
cloudiness, high temperature, low humidity and virtually
no rainfall during the summer. The winter is mild, with a
mean air temperature of about 10 C during the coldest
month (January). Mean annual temperature was 18.2 C in
2004 and precipitation was 325 mm. These values are close
to climatic means determined over 20 years at a nearby
weather station (mean air temperature was 18.3 ± 0.9 C
and mean precipitation was 355 ± 90 mm). The corre-
sponding value of reference crop evapotranspiration (ETo),
calculated with the FAO Penman–Monteith equation (Allen
et al. 1998), is 1180 ± 60 mm. The soil texture was a deep
silt-clay-loam with low available potassium and organic mat-
ter concentrations and high phosphorus concentration. Elec-
trical conductivity (EC) of the saturation extract was
1.4 dS m1 in 2004. The bulk density of the surface layer
was 1.4 ± 0.1 g cm3 and available water-holding capacity
of soil was about 0.18 mm1.
Irrigation regimes
In 2000, two irrigation treatments, FI and sustained DI
were applied to the experimental plots until the end of
2004, following a completely randomized statistical design
with three replicates per treatment and 12 trees per
 SEASONAL CHANGES IN ALMOND PHOTOSYNTHETIC PARAMETERS 377

I II - III IV V
100

h
Relative value (%)

th

wt
80 w

ro
wth
ro leaves fall
kg

sg
un

gro

he
Tr

Flowering
Figure 1. Schematic representation of

nc
60

uit

ra
P. dulcis tree developmental Stages I–V. The

Fr

gb
un
40 curves show the relative cumulative values of

Yo
Reserves
fruit growth, young branch growth and fruit
20
dry mass accumulation (kernel-filling), with

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Kernel-filling
0 respect to their maximum, for the cultivar
studied (cv Marta) in the FI regime in
Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec
southern Spain, as a function of DOY.
Vertical lines delimit the beginning and the
DOY: 31 59 90 120 151 181 212 243 273 304 334 365 end of each developmental stage.

replicate. Trees were irrigated by a single drip line equipped Measurements of leaf N concentration and leaf area
with six pressure compensated drippers (4 l h1 per dripper)
Foliar N concentration was measured from April to
per tree.
September 2004 in the fully expanded leaves of non-fruiting
In the FI plots, an amount of water corresponding to
branches (current-year branches) and fruiting branches
100% of the standard crop evapotranspiration, ETc, was
(1-year-old branches) – hereafter nf- and f-leaves, respec-
supplied weekly. We calculated ETc as the product of
tively – in the two irrigation treatments. Samples consisted
ETo and the crop coefficient for almond trees (Doorenbos
of at least 12 leaves in each class taken from the upper third
and Pruitt 1977), and corrected with a localization factor
of sun-exposed branches. Leaf area was measured with an
(Fereres et al. 1982). During the first 3 years (2000–2002),
area meter (Delta-T Image Analysis System (DIAS),
the trees in the DI plots were irrigated following a sustained
Delta-T Devices, Cambridge, UK).
DI strategy, receiving 60% of ETc in the entire irrigation
Mean leaf N concentration was determined after Kjel-
season. In 2003 and 2004, the water supply was further
dahl digestion of leaves that had been oven-dried at
restricted such that the trees received about 40% of ETc.
80 C for 48 h, weighed and ground. For each class of
The EC of irrigation water was 1.2 dS m1 with chloride
leaves, leaf N concentration is expressed per unit leaf area
and sodium concentrations of 4.6 and 5.41 meq l1, respec-
(Na, in g m2) and per unit leaf dry mass (DW) (Nw, in g
tively. Trees were fertilized with 35, 35, 57 kg ha1 year1
g1). Leaf dry mass per unit leaf area (Wa) is expressed in
of N, P, K and were managed according to current com-
g m2.
mercial practices (i.e., a routine pesticide program was
maintained, pruning was applied manually in December,
and all weeds were removed from the orchard). Gas exchange measurements
Leaf gas exchange was measured from April to September on
Measurement period
trees in the FI and DI irrigation regimes. The criteria adopted
In 2004, leaf characteristics were measured from the begin- for selecting leaves on the two classes of branches were similar
ning of April (day of year (DOY) 97, when fruits had to those described for the leaf N concentration measure-
reached 92% of their final size) until post-harvest, corre- ments. Only healthy branches were selected from the periph-
sponding to the period of reserve accumulation in woody ery of the tree crown. Area-based leaf net CO2 assimilation
structures. Fruit harvest occurred in mid-August (DOY (A, lmol m2 s1), transpiration rate (E, mmol m2 s1),
220) for trees in the DI treatment and about 10 days later stomatal conductance to water vapor (gs, mmol m2 s1)
for trees in the FI treatment. and intercellular CO2 concentration (Ci, lmol mol1) were
Figure 1 gives a schematic representation of the stages of measured with a portable gas exchange system CIRAS2
tree development as a function of DOY for trees in the FI (PP Systems, Hitchin, Hertfordshire, UK). The desired pho-
regime. The curves of relative cumulative values of fruit tosynthetic photon flux (I, lmol m2 s1) was provided by
growth, young branch growth and fruit dry mass accumula- an internal red/blue LED light source (PC 069-1). A CIRAS2
tion (kernel-filling) were normalized against their maximum injection system controlled the ambient CO2 concentration
values. Before the experimental period (February 2004), (Ca, lmol mol1) in the chamber by adjusting the flow of
mean trunk diameter of the trees was 14.08 ± 0.28 cm in CO2 from a CO2 cylinder, and also controlled the leaf tem-
FI and 12.02 ± 0.15 cm in DI, and canopy height was perature (Tl) in the chamber. Air temperature (Ta) and actual
3.56 ± 0.09 m in FI and 3.19 ± 0.07 m in DI. At the end vapor pressure of the chamber air were continuously moni-
of the 2003 growing season, crown leaf area was tored by the CIRAS2. Measurements were made at periodic
68.11 ± 2.48 m2 tree1 in FI and 41.94 ± 3.58 m2 tree1 intervals ( 7 to 10 days) throughout the growth season on
in DI. at least six leaves (one leaf per branch, two branches per tree

TREE PHYSIOLOGY ONLINE at http://www.treephys.oxfordjournals.org

378 NORTES, GONZALEZ-REAL, EGEA AND BAILLE
and three trees per irrigation treatment). All leaf gas
exchange measurements were made between 1000 and
1300 h (local time).
Light-saturated leaf net CO2 assimilation (Am) and max-
imum stomatal conductance (gsm) were measured at I
 1400 lmol m2 s1, near constant Ca ( 360 lmol
mol1) and Tl ( 30 C). Area- and dry-mass-based values
of Am are denoted as Ama (lmol m2 s1) and Amw (lmol
g1 s1), respectively. Light-saturated, PNUEm, was com-
puted as the ratio Amw/Nw (lmol g1 s1).
Leaf potential net CO2 assimilation or maximum meta-
bolic capacity (Ap) was measured from April to September
in trees in both FI and DI regimes, at saturating I ( 1400
lmol m2 s1) and saturating Ca ( 1400 lmol mol1),
and at near optimum Tl (35 C). This Tl value was derived
in spring from the measurements of Ap in FI leaves over a
wide range of Tl (18–42 C).
Tree water status
Every 2 weeks, we measured predawn leaf water potential
(Wpd) with a pressure chamber (Model 3000, Soil Moisture
Equipment, Santa Barbara, CA) on mature leaves (12
leaves per irrigation treatment, i.e., six trees and two leaves
per tree) taken before dawn from the upper third of the
branches. Data for the f- and nf-leaves were pooled.
Statistical analysis
We analyzed (1) leaf N concentration (area- and dry-mass-
based) as a dependent variable against Wa and (2) the pho-
tosynthetic parameters (area- and dry-mass-based) as
dependent variables against Wa and against leaf N concen-
tration (area- and dry-mass-based) by using linear regres-
sion. Significant differences between slopes and non-zero
intercepts were evaluated by analyses of covariance, by com-
paring (1) FI and DI per each leaf class, (2) FI leaves and DI
leaves with pooled data for the two leaf classes, (3) the two
leaf classes with pooled data from FI and DI for each leaf
class and (4) all datasets. To discriminate the main treatment
effects on leaf structural properties and on photosynthetic
parameters analysis of variance (ANOVA) was used. All
analyses were performed using Statgraphics software
(Statgraphics Plus for Windows Version 4.1).
Results
Climate conditions
Maximum absolute values of air temperature (Ta) and
vapor pressure deficit (VPD) increased rapidly from spring
(23 C and 2.2 kPa, respectively) to summer (36.6 C and
4.4 kPa, respectively) (Figure 2A), corresponding to the
increase in the daily integral of global solar radiation over
the same period (Figure 2C). During sunny days, mean val-
ues of Ta and VPD remained below 18 C and 1.1 kPa,
respectively, in spring and below 29 C and 2.1 kPa,
TREE PHYSIOLOGY VOLUME 29, 2009
respectively, in summer (Figure 2B). The calculated refer-
ence crop evapotranspiration (ETo) was typical for the loca-
tion, with maximum daily values in summer of about
8 mm day1 (Figure 2D). Because of the high amount of
precipitation in spring 2004 (Figure 2D), irrigation started
on May 3 in all plots. Practically no rain occurred from
the end of June until post-harvest (September). For the
entire irrigation season (May until the end of November),
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the total amount of irrigation water was 410 and 170 mm
for FI and DI, respectively, and rainfall was 355 mm.
Seasonal variation in predawn leaf water potential
In both irrigation treatments, predawn leaf water potential
Wpd was 0.30 ± 0.03 MPa during the active vegetative
growth period (i.e., Stages II and III; Figures 1 and 3), indi-
cating high soil water availability during this period. The FI
trees maintained Wpd within a small range from the onset of
the kernel-filling stage (0.35 ± 0.02 MPa) until post-
harvest (0.45 ± 0.01 MPa) compared with the range of
variation observed in DI trees over the same period (from
0.47 ± 0.05 to 0.83 ± 0.06 MPa, respectively).
Seasonal trends in leaf structural traits
The seasonal trends in leaf dry mass per unit area (Wa)
(Figure 4A) and area-based nitrogen concentration (Na)
(Figure 4B) were characterized by two seasonal peaks that
were more marked in nf-leaves than in f-leaves. For both
leaf traits, the first peak occurred near DOY 140–145 (onset
of the kernel-filling stage), whereas the absolute maximum
was reached just before harvest (DOY 218–220). Dry mat-
ter accumulated in the leaf blade except for two periods
coinciding with the first half of kernel-filling and the onset
of reserve accumulation (Figure 4A). Over time, Wa
showed a greater increase in FI trees (about 36% in both
leaf classes) than in DI trees (21% and 29% in f- and nf-
leaves, respectively) (Figure 4A). Independently of irriga-
tion treatment, Wa was generally greater in nf-leaves than
in f-leaves. The FI trees maintained differences between
the two leaf classes when Wa was averaged over the growth
season (P = 0.02), whereas the corresponding differences
in DI trees were significant only across Stages IV and V
(P  0.04). Restriction of water supply moderately – but
significantly – reduced Wa in nf-leaves (about 10%) during
post-harvest compared with nf-leaves of FI trees
(P < 0.01), but the DI treatment did not influence Wa in
f-leaves during the growing season. Values of Wa were
highly correlated between f- and nf-leaves within each irri-
gation regime (R2 > 0.90, data not shown).
Area-based leaf N concentration (Na) was significantly
higher in nf-leaves than in f-leaves (Figure 4B) from the
onset of kernel-filling stage onward in both irrigation treat-
ments. In FI trees, Na averaged over the measurement per-
iod was 27% higher in nf-leaves (3.1 ± 0.3 g m2) than in
f-leaves (P < 0.01), a greater difference than that observed
for Wa (17%). In DI, the difference in Na among leaf classes
 SEASONAL CHANGES IN ALMOND PHOTOSYNTHETIC PARAMETERS 379

40 5 40 5
Tam
A B VPDm
4 4
30 30

VPDmx (kPa)

VPDm (kPa)
Tamx (ºC) 3 3

Tam (ºC)
20 20
2 2

10 10

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VPDmx 1 1

Tamx
0 0 0 0
35
8
30 C D 140
7

Rainfall (mm month 1)

120
Σ G (MJ m 2 d 1)

25 6

ETo (mm d 1)
100
20 5
80
15 4
60
10 3
40
2 ETo
5 20
1 Rainfall
0
50 100 150 200 250 300 350 50 100 150 200 250 300 350
DOY DOY

Figure 2. (A) Maximum diurnal values of air temperature (Tamx) and air vapor pressure deficit (VPDmx). (B) Mean diurnal values of
air temperature (Tam) and VPD (VPDm). (C) The integral of global solar radiation (RG). (D) The reference crop evapotranspiration
(ETo) and monthly values of rainfall for the 2004 growing season. Mean daytime values are for the period from sunrise to sunset.
Abbreviation: DOY, day of year.

0.0 the amount of N in nf-leaves was 15% higher in FI trees

FI
DI than in DI trees (P < 0.01). The N concentration in
f-leaves was not significantly affected by irrigation regime.
0.4 The seasonal variations in Wa and Na resulted in a sus-
Ψpd (MPa)

tained decrease in Nw (=Na/Wa) over time (Figure 4C).

0.8
Stages II-III Stage IV
1.2
75 125 175
DOY
Figure 3. Time course of (Wpd) in P. dulcis trees through the
2004 growing season. Vertical bars indicate standard deviations.
Each value is the mean of 12 measurements per irrigation
treatment (pooled data of f- and nf-leaves). Vertical dotted lines
delimit the beginning and the end of each developmental stage
(cf. Figure 1). Abbreviations: f- and nf-leaves were obtained
from 1-year-old fruiting (f) and current-year non-fruiting (nf)
shoots, respectively; FI, full irrigation; DI, sustained deficit
irrigation at 50% of standard crop evapotranspiration through-
out the growing season and DOY, day of year.
was much lower (about 13% higher in nf-leaves)
(P = 0.01) and was of the same order of magnitude as that
observed for Wa. Thus, on average over the growth season,
Differences in Nw between leaf classes were observed in
FI trees only during Stages IV and V (14% higher in nf-
leaves than in f-leaves, P = 0.03), i.e., about twofold lower
Stage V than those observed for Na. The nf-leaves maintained
a higher Nw over the growth cycle in the FI treatment
225 275
(0.0274 ± 0.003 g g1) than in the DI treatment
(P = 0.02), a pattern similar to that observed for Na.
Regression analyses: N versus Wa relationships
The results of the regression analyses between leaf N con-
centration (Na or Nw) and Wa are shown in Table 1, for
all combinations of leaf class and irrigation treatment over
the entire growth cycle. There were significant relationships
between Na and Wa in nf-leaves in both FI and DI treat-
ments (R2 = 0.61 and R2 = 0.43, respectively), but not
in f-leaves (R2  < 0.01) (Table 1). For the pooled dataset,
there was a highly significant relationship (P < 0.001)
between Na and Wa (R2 = 0.37), as well as between Nw
and Wa (R2 = 0.29). For each developmental stage, signif-
icant relationships (P < 0.001) were obtained when

TREE PHYSIOLOGY ONLINE at http://www.treephys.oxfordjournals.org

380 NORTES, GONZALEZ-REAL, EGEA AND BAILLE

irrespective of leaf class and irrigation regime. No significant

160 nf-FI f-FI nf-DI f- DI
A differences were observed between the slopes for the irriga-
140 tion treatments, but the non-zero intercepts differed signifi-
Wa (g m 2 )

cantly (Table 1). For the pooled data for FI and DI in

120
each leaf class, the relationships between Nw and Wa were
100 significant for both nf- (R2 = 0.41, P < 0.005) and f-leaves
80 (R2 = 0.83, P < 0.001). The data for f-leaves in the FI and
DI treatments displayed a unique relationship, suggesting
60

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Stages II-III Stage IV Stage V
that the DI treatment did not modify the partitioning of N
4 in f-leaves, whereas the nf-leaves had higher Nw in the FI
B treatment than in the DI treatment for a given Wa (Figure 6).
3
Na (g m 2 )

Seasonal trends in photosynthetic attributes

2 Light-saturated photosynthetic attributes showed a non-
uniform trend over the growth cycle (Figure 7A–C), with
1 most of the attributes tending to reach a more or less clear
Stages II-III Stage IV Stage V
maximum. Area-based Am was highest during Stages II and
0.035 III, decreasing rapidly after the onset of Stage IV (Figure
100 150 200 250
X Data
C 7A). In both irrigation regimes, the differences in Ama
0.030 between leaf classes were not significant, except in the DI
treatment during Stage IV when nf-leaves had significantly
Nw (g g 1)

0.025 higher values than f-leaves (20.0 ± 1.4 versus

0.020
Stages II-III Stage IV
0.015
100 150 200
DOY
Figure 4. Seasonal trends for (A) area-based leaf dry mass (Wa),
(B) area-based leaf N concentration (Na) and (C) mass-based
leaf N concentration (Nw) in the FI (open symbols) and DI
(closed symbols) regimes, in P. dulcis f-leaves (circles) and
nf-leaves (triangles) as a function of DOY. Pooled standard
deviations of the mean (not shown for clarity) were typically
± 5 g m2, ± 0.015 g m2 and ± 0.0012 g g1 for Wa, Na and
Nw, respectively. Vertical dotted lines delimit the beginning and
the end of each developmental stage (cf. Figure 1). Abbrevia-
tions: f- and nf-leaves were obtained from 1-year-old fruiting (f)
and current-year non-fruiting (nf) shoots, respectively; FI, full
irrigation and DI, sustained deficit irrigation at 50% of standard
crop evapotranspiration throughout the growing season.
pooling data from Stages IIIII (Figure 5A), Stage IV (Fig-
ure 5B) and Stage V (Figure 5C). Forcing the relationships
through the origin provided a slope (i.e., the mean Nw over
a given stage) of 0.0285 and 0.0211 g g1 for Stages II, III
and V, respectively, indicating a substantial decrease in Nw
for both leaf classes and in the two irrigation regimes dur-
ing the post-harvest stage. For comparison, the relationship
reported by DeJong and Doyle (1985) for Prunus persica
(L.) Batsch (Na = 0.027 Wa + 0.002) is shown in Figure
5A–C.
In contrast to Na, Nw decreased with increasing Wa
in both leaf classes (Figure 6), with significant relationships
(0.60 < R2 < 0.93) over the entire growth season,
17.6 ± 0.5 lmol m2 s1) (P < 0.01). The f-leaves had
similar Ama in both irrigation treatments, whereas the nf-
leaves had significantly lower Ama in the DI treatment than
Stage V in the FI treatment during Stages II, III and V (17% and
38%, respectively). Mean Ama was not significantly affected
250
by irrigation regime during the kernel-filling stage, regard-
less of leaf class.
Mass-based Am tended to decrease during the growing
season (Figure 7B), declining in FI leaves during Stages II
and III (from about 0.30 ± 0.05 to 0.22 ± 0.02 lmol g1
s1) and then reaching a plateau until the first half of
kernel-filling stage, coinciding with the decline in Wa
(Figure 4A). In contrast, Amw was rather constant in DI
leaves over the same period ( 0.20 lmol g1 s1);
however, after harvest, it declined to low values ( 0.10
lmol g1 s1, pooled data for both irrigation regimes).
The relative decline over the growing season was greater
for Amw ( 61%) than for Nw (37%), and the differences
in Amw between leaf classes and between irrigation regimes
were significantly reduced during the kernel-filling stage.
The seasonal trend in gsm (Figure 7C) showed maximum
absolute values during the kernel-filling stage (323 ± 35
and 273 ± 20 mmol m2 s1 for nf-leaves in FI and DI
trees, respectively). Peak values were more marked in FI
trees than in DI trees, because of lower variability in gsm
in the former. After reaching a maximum, gsm more or less
paralleled the downward trend observed for Ama. The mag-
nitude of this decrease was lower in FI trees than in DI
trees, with a mean reduction in gsm for the two leaf classes
of 85 mmol m2 s1 in the FI treatment and 153 mmol
m2 s1 in the DI treatment for the period DOY 154–
245. The greatest differences in gsm between irrigation
regimes were observed during the post-harvest stage.

TREE PHYSIOLOGY VOLUME 29, 2009

 SEASONAL CHANGES IN ALMOND PHOTOSYNTHETIC PARAMETERS 381

Table 1. Linear regression coefficients for Na versus Wa and Nw versus Wa for the two leaf classes (f- and nf-leaves) and the two
irrigation regimes (FI and DI). Significance values: ns, not significant; *P < 0.05; **P < 0.01; and ***P < 0.001. For each leaf
class · irrigation treatment, different letters within the same column indicate significant differences between the non-zero intercepts and
between the slopes (P < 0.05). Abbreviations: f- and nf-leaves were obtained from 1-year-old fruiting (f) and current-year non-fruiting
(nf) shoots, respectively; FI, full irrigation and DI, sustained deficit irrigation at 50% of standard crop evapotranspiration throughout
the growing season. Units: Wa, g m–2; Na, g m–2 and Nw, g g–1.
Na = bo + b1 Wa Nw = bo + b1 Wa
2
bo b1 R bo b1 R2

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f-leaves FI 2.131***a 0.0015ns a 0.03 ns 0.0495***a 0.00026***a 0.93***
DI 2.808**a 0.0049ns a 0.06 ns 0.0521***a 0.00029**a 0.71**
nf-leaves FI 1.744**a 0.0117**a 0.61** 0.0423***a 0.00013***a 0.78***
DI 1.467*b 0.0107*a 0.43* 0.0377***b 0.00012**a 0.60**
f + nf-leaves FI 0.952*a 0.0165***a 0.48** 0.0362***a 0.00010*a 0.26*
DI 1.508**a 0.0094*a 0.23* 0.0388***a 0.00014**a 0.43**
FI + DI f-leaves 2.338***a 0.0004ns a 0.00ns 0.0503***a 0.00027***a 0.83***
nf-leaves 1.423**b 0.0128**b 0.40** 0.0385***b 0.00011**b 0.41**
All datasets All datasets 1.105** 0.0142*** 0.37*** 0.0367*** 0.00011*** 0.29***

The ratio of intercellular CO2 concentration to ambient
CO2 concentration (Ci/Ca) was lowest during the active veg-
etative growth period (data not shown). The ratio tended to
increase from Stages II and III (mean value in FI leaves:
0.52 ± 0.07) until the second half of kernel-filling stage
and then remained more or less constant (mean value in
FI leaves beyond Stage V: 0.61 ± 0.04).
Area-based leaf potential net CO2 assimilation (Apa)
exhibited a seasonal trend similar to that of Ama, with high-
est rates during Stages II and III (Figure 7D) and a decrease
over the growing season that was independent of leaf class
and irrigation regime. The differences in Apa among all
combinations of leaf class and irrigation treatment were
similar to or lower than in Ama.
Analyses of the relationships between Am and Wa
and between Am and leaf N concentration
Regression analyses revealed highly significant relationships
between Am and Wa, and between Am and Nw (P < 0.001)
for all combinations of leaf class and irrigation treatment,
as well as for the pooled datasets. Two examples of Amw
versus Wa relationships are given in Figure 8A and B.
For the pooled data for f- and nf-leaves per irrigation treat-
ment (relationships for FI leaves and DI leaves, Figure 8A),
no significant differences between irrigation regimes were
observed in slopes or in non-zero intercepts. For the pooled
data for the FI and DI treatments per leaf class (relation-
ships for f- and nf-leaves, Figure 8B), there were significant
differences in slopes and non-zero intercepts between leaf
classes. The relationships between Amw and Wa (Figure
8B), and between Nw and Wa (Figure 6) were similar. For
all pooled datasets, Wa explained about two-thirds of the
seasonal variability in Amw (R2 = 0.65, P < 0.001). The
regression analysis for Ama versus Wa yielded lower R2
values than for Amw versus Wa.
Seasonal changes in Amw and Ama were poorly correlated
to changes in Na (R2 < 0.10), but were correlated with Nw
(0.69 < R2 < 0.90,) with P < 0.001 for the linear regres-
sions between either Amw or Ama versus Nw, for all combi-
nations of leaf class and irrigation regime. Two examples of
Am versus Nw relationships are shown in Figure 9A (Ama
versus Nw for FI leaves and DI leaves, pooled data of
f- and nf-leaves) and Figure 9B (Amw versus Nw for
f- and nf-leaves, pooled data of FI and DI).
PNUE
The seasonal trend in light-saturated PNUEm reached a
plateau from Stages II and III until the second half of
kernel-filling stage (Figure 10A) and then decreased dra-
matically, reaching a minimum after harvest. Regression
analyses of the PNUEm versus Wa, PNUE versus Na, and
PNUE versus Nw relationships indicated that PNUEm
was strongly correlated with Wa and Nw but loosely corre-
lated with Na (data not shown). The relationship between
PNUEm and Nw (Figure 10B) shows that, for a fixed value
of Nw, PNUEm was generally higher in f-leaves than in nf-
leaves, irrespective of irrigation regime. Pooling all the data
revealed that PNUEm was more closely correlated with Wa
(R2 = 0.78; Figure 10C) than with Nw (R2 = 0.49). The
three outliers in Figure 10C correspond to the post-harvest
period in the DI regime.
Nitrogen-use efficiency versus photosynthetic N
An estimate of the amount of leaf N concentration diverted
to the photosynthetic apparatus (i.e., dry-mass-based pho-
tosynthetic N, Nwp) was derived from Eq. (1) (Field and
Mooney 1983), based on the pooled data for FI and DI
for each leaf class
Amw ¼ mðN w  N wo Þ ¼ mN wp ; ð1Þ
where Nwo is the residual value of leaf N concentration
when Amw = 0 and the slope m represents the mean
PNUE related to photosynthetic leaf N concentration,

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382 NORTES, GONZALEZ-REAL, EGEA AND BAILLE

A 0.035
f+nf-leaves, FI+DI
3.5 0.030
R2 = 0.78
3.0

Nw (g g )
1
Na (g m 2 )

0.025
2.5 R2 = 0.83
0.020
2.0 stages II-III R2 = 0.60

y = 0.0285x 0.015
1.5

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R2 = 0.34 f(FI+DI) nf-FI nf-DI
1.0 0.010

B 50 75 100
Wa (g m 2 )
125 150

3.5 f+nf-leaves, FI+DI

3.0 Figure 6. Relationship between leaf N concentration (Nw) and

Na (g m 2 )

leaf dry mass per unit leaf area (Wa) in P. dulcis nf-leaves in the
2.5
FI and DI irrigation treatments and for f-leaves based on pooled
2.0 data from the FI and DI treatments. Regression parameters and
stage IV significance level of slopes and non-zero intercepts are given
1.5 y = 0.0260x in Table 1. Abbreviations: f- and nf-leaves were obtained from
R2 = 0.66 1-year-old fruiting (f) and current-year non-fruiting (nf) shoots,
1.0 respectively; FI, full irrigation and DI, sustained deficit irriga-
C tion at 50% of standard crop evapotranspiration throughout the
growing season.
3.5 f+nf-leaves, FI+DI

3.0
Na (g m 2 )

stage V between leaf classes, in contrast to what was observed for

2.5 y = 0.0211x
R2 = 0.75
2.0
1.5
1.0
25 50 75 100 125
2
Wa (g m )
Figure 5. Relationship, forced through the origin, between leaf
N concentration per unit leaf area (Na) and leaf dry mass per
unit leaf area (Wa) for different stages of development of
P. dulcis trees (A, B and C) (pooled data for f- and nf-leaves in
the FI and DI treatments). Standard deviations of the mean
(data not shown) were typically ± 5 g m2 and ± 0.015 g m2
for Wa and Na, respectively. The dashed line is the linear
relationship reported by DeJong and Doyle (1985) for P. persica
(y = 0.027x + 0.002). In (A), (B) and (C), the continuous lines
correspond to the respective linear regressions. Abbreviations:
f- and nf-leaves were obtained from 1-year-old fruiting (f) and
current-year non-fruiting (nf) shoots, respectively; FI, full
irrigation and DI, sustained deficit irrigation at 50% of standard
crop evapotranspiration throughout the growing season.
i.e., PNUEmp = Amw/Nwp. Assuming that Eq. (1) is valid
over the entire range of Nw, and that, for a given leaf
class, Nwo is constant throughout the growing season
and is independent of the irrigation regime (Figure 9B),
the regression analysis yielded for f-leaves: Nwo = 0.0127
± 0.004 g g1 and PNUEmp = 15.62 ± 1.78 lmol g1
s1; and for nf-leaves: Nwo = 0.0153 ± 0.005 g g1,
and PNUEmp = 16.14 ± 2.38 lmol g1 s1. Thus, the
values of Nwo differed significantly between leaf classes
(P < 0.05), whereas PNUEmp did not differ significantly
PNUEm (Figure 10B).
The trend in photosynthetic N as a fraction of total leaf
N concentration (fpN = Nwp/Nw) indicated that, in both
irrigation regimes, the f-leaves had a high fpN during Stages
II and III ( 0.60 on DOY 90) compared with nf-leaves
150 ( 0.50 and 0.40 in FI and DI, respectively) (Figure 11).
Thereafter, fpN tended to decline in both leaf classes, the
greatest decrease occurring during the second half of the
kernel-filling stage. In the post-harvest stage, fpN ranged
from 0.25 (nf-leaves in DI) to 0.40 (f-leaves in FI).
Discussion
Effects of irrigation treatment on leaf structural traits
in fruiting versus non-fruiting shoots
Our study highlighted two main characteristics of the sea-
sonal pattern of leaf structural traits. First, there was a close
similarity in the seasonal patterns of Wa, Na and Nw
between FI and DI trees (Figure 4A–C). The greatest sim-
ilarity was in Nw, which exhibited a decrease of about 40%
throughout the season, independently of leaf class and irri-
gation treatment. This reduction was greater than that
reported for other temperate deciduous trees (Niinemets
et al. 2004) but similar to that found in almond leaves by
Weinbaum and Muraoka (1986). There are several possible
explanations for this seasonal pattern: (1) dilution of N in
the leaf blade, because of the accumulation of non-N com-
pounds; (2) N allocation to fruits and woody structures and
(3) changes in N partitioning between metabolic and struc-
tural components within the leaf (Yasumura et al. 2006).
Second, independently of irrigation treatment, the f-leaves

TREE PHYSIOLOGY VOLUME 29, 2009

 SEASONAL CHANGES IN ALMOND PHOTOSYNTHETIC PARAMETERS 383

30 0.4
nf-FI nf-FI
25
A f-FI B f-FI
nf-DI nf-DI

Amw (μmol g s )
0.3

Ama (μmol m s )

1
1
f-DI f-DI
20

1
2
15 0.2

10
0.1
5

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Stages II-III Stage IV Stage V Stages II-III Stage IV Stage V
0 0.0
80
400 C D 70
gsm (mmol m s )

Apa (μmol m s )
60
1

1
300
2

2
50
40
200
30

100 20
10
Stages II-III Stage IV Stage V Stages II-III Stage IV Stage V
0 0
100 150 200 250 100 150 200 250

DOY DOY

Figure 7. Seasonal trends in (A) area-based (Ama) and (B) dry-mass-based (Amw) light-saturated net CO2 assimilation, (C) maximum
stomatal conductance (gsm) and (D) potential net CO2 assimilation (Apa) in FI (open symbols) and DI (closed symbols) regimes, in
P. dulcis f-leaves (circles) and nf-leaves (triangles) as a function of DOY. Pooled standard deviations of the mean (not shown for clarity)
were typically close to (A) ± 2.0 lmol m2 s1, (B) ± 0.052 lmol g1 s1, (C) ± 47 mmol m2 s1 and (D) ± 4.2 lmol m2 s1.
Vertical dotted lines delimit the beginning and the end of each developmental stage (cf. Figure 1). Abbreviations: f- and nf-leaves were
obtained from 1-year-old fruiting (f) and current-year non-fruiting (nf) shoots, respectively; FI, full irrigation and DI, sustained deficit
irrigation at 50% of standard crop evapotranspiration throughout the growing season.

0.4 0.4
A FI
DI
B f-leaves
nf-leaves
Amw (μmol g 1 s 1 )

Amw (μmol g 1 s 1 )
0.3 0.3

0.2 0.2

0.1 0.1

0.0 0.0
50 75 100 125 150 50 75 100 125 150

Wa (g m 2 ) Wa (g m 2 )

Figure 8. Relationships between mass-based maximum leaf net CO2 assimilation (Amw) and leaf dry mass per unit area (Wa) in
P. dulcis trees (A) in the FI and DI regimes, using pooled data from f- and nf-leaves and (B) for f- and nf-leaves, using pooled data from
FI and DI regimes. Regression lines in (A) y = 0.475  0.0028x (R2 = 0.73, P < 0.001) and y = 0.470  0.0030x (R2 = 0.64,
P < 0.001) for FI and DI, respectively, and in (B) y = 0.623  0.0046x (R2 = 0.87, P < 0.001) and y = 0.472  0.0027x
(R2 = 0.72, P < 0.001) for f- and nf-leaves, respectively. Abbreviations: f- and nf-leaves were obtained from 1-year-old fruiting (f) and
current-year non-fruiting (nf) shoots, respectively; FI, full irrigation and DI, sustained deficit irrigation at 50% of standard crop
evapotranspiration throughout the growing season.

had lower Wa, Na and Nw than the nf-leaves (Figure 4A–C). nf-leaves, which had significantly lower Na in DI trees than
The DI regime did not significantly affect Wa in f-leaves but in FI trees, whereas Na in f-leaves was little affected by the
decreased Wa in nf-leaves by 10–15%, mainly from pre-har- DI treatment (Figure 4B). A similar conclusion can be
vest onward (Figure 4A). Stress conditions have been drawn for Nw (Figure 4C). Variations in Na (=WaNw)
reported to affect leaf N concentration (Reich et al. may result from variations in Wa, in Nw, or in both. A sin-
1989). This was confirmed in our study, but only for the gle relationship characterized the whole dataset when the

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384 NORTES, GONZALEZ-REAL, EGEA AND BAILLE

30 0.4
FI
DI
A f-leaves
nf-leaves
B
25

Ama (μmol m 2 s 1 )

Amw (μmol g 1 s 1 )
0.3
20

15 0.2

10
0.1
5

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0 0.0
0.010 0.015 0.020 0.025 0.030 0.035 0.010 0.015 0.020 0.025 0.030 0.035

Nw (g g 1) Nw (g g 1)

Figure 9. Relationships (A) between area-based maximum leaf net CO2 assimilation (Ama) and mass-based leaf N concentration (Nw)
in P. dulcis trees in the FI and DI regimes, using pooled data from f- and nf-leaves and (B) between dry-mass-based Am (Amw) and Nw
for the f- and nf-leaves, using pooled data from FI and DI regimes. Regression lines in (A) y = 4.01 + 868.91x (R2 = 0.76,
P < 0.001) and y = 13.05 + 1217.9x (R2 = 0.80, P < 0.001) for FI and DI, respectively, and in (B) y = 0.199 + 15.62x
(R2 = 0.89, P < 0.001) and y = 0.247 + 16.04x (R2 = 0.80, P < 0.001) for f- and nf-leaves, respectively. Abbreviations: f- and
nf-leaves were obtained from 1-year-old fruiting (f) and current-year non-fruiting (nf) shoots, respectively; FI, full irrigation and DI,
sustained deficit irrigation at 50% of standard crop evapotranspiration throughout the growing season.

12 12
A nf-FI
f-FI B
PNUEm (μmol g s 1 )

10 10
PNUEm (μmol g s )
1

nf-DI
1

8 f-DI 8
Figure 10. (A) Seasonal trend in
6 6 PNUEm in P. dulcis trees. Symbols as
in Figure 4. Pooled standard devia-
4 4 tion of the mean was typically close
to ±1.0 lmol g1 s1. The (B and C)
2 2 f-leaves relationships between PNUEm and
nf-leaves
Stages II-III Stage IV Stage V Nw and between PNUEm and Wa,
0 0 respectively. Regression lines showed
100 150 200 250 0.010 0.015 0.020 0.025 0.030 0.035 in (B) for f-leaves (1.21 + 346.9Nw,
DOY Nw (g g 1 ) R2 = 0.70, P < 0.001) and nf-leaves
12 (3.8 + 396.0Nw, R2 = 0.60,
C nf-FI P < 0.001), using pooled data from
s 1)

10 f-FI FI and DI regimes, and in (C) for all

nf-DI datasets (15.14  0.080Wa,
1

8 f-DI
R2 = 0.78; P < 0.001) excluding the
PNUEm (μmol g

three outliers corresponding to the

6
post-harvest period in the DI regime.
4 Postharvest Abbreviations: f- and nf-leaves were
DI regime obtained from 1-year-old fruiting (f)
2 and current-year non-fruiting (nf)
shoots, respectively; FI, full irrigation;
0 DI, sustained deficit irrigation at 50%
50 75 100 125 150 175 of standard crop evapotranspiration
throughout the growing season and
Wa (g m 2 ) DOY, day of year.

Na versus Wa relationship was analyzed separately for each corresponding values in nf-leaves, implying that f-leaves
stage of development (Figure 5A–C), but the slope value experienced a greater decrease in Nw for a given Wa and
(giving the mean Nw over a stage) shifted as the growing had a lower nitrogen concentration at high Wa (Figure 6).
season progressed, indicating that the Na versus Wa rela- This may be ascribed to a higher depletion of N in the leaf
tionship is stage-dependent in almond trees, in contrast to blade to cope with the demand of the fruits during the
nectarine trees (Rosati et al. 1999). kernel-filling stage and that of the perennial organs after
Independently of irrigation treatment, the presence of harvest. A unique linear Nw versus Wa relationship was
fruits near the leaves increased the slope and the intercept found for f-leaves in the FI and DI treatments, whereas
of the Nw versus Wa relationship compared with the this relationship differed significantly between irrigation

TREE PHYSIOLOGY VOLUME 29, 2009

 SEASONAL CHANGES IN ALMOND PHOTOSYNTHETIC PARAMETERS
0.8
nf-FI
f-FI
0.6 nf-DI
f-DI
fpN
0.4
0.2
Stages II-III Stage IV Stage V
0.0
100 150 200 250
DOY
Figure 11. Seasonal trend in photosynthetic-N fraction, fpN
(=Nwp/Nw) in P. dulcis trees. Same symbols as Figure 4. Pooled
standard deviation of the mean (data not shown) was typi-
cally ± 0.040. Abbreviations: f- and nf-leaves were obtained
from 1-year-old fruiting (f) and current-year non-fruiting (nf)
shoots, respectively; FI, full irrigation; DI, sustained deficit
irrigation at 50% of standard crop evapotranspiration through-
out the growing season and DOY, day of year.
treatments for nf-leaves (Figure 6), suggesting that, in the
DI regime, leaf structural adjustments operated to maintain
a conservative N status in the leaves of fruit-bearing shoots,
but to the detriment of N resources allocated to vegetative
shoots.
Effects of irrigation treatment on leaf physiologic traits in
fruiting versus non-fruiting shoots
Almond species are known to exhibit high rates of photo-
synthesis among woody Rosaceae (DeJong 1983, Wullsch-
leger 1993, Higgins et al. 1992, Lakso 1994, González-Real
and Baille 2000). We observed maximum absolute Ama val-
ues close to 24 lmol m2 s1 (Figure 7A), consistent with
the values reported for almond leaves (DeJong 1983,
Higgins et al. 1992, Matos et al. 1997, De Herralde et al.
2003, Rouhi et al. 2007).
Independently of the irrigation regime, leaves of fruiting
and non-fruiting shoots exhibited high Ama during the
active vegetative growth period when the values of gsm were
below their maximum (Figure 7A–C), therefore maintain-
ing a high intrinsic water-use efficiency, as observed by
Heilmeier et al. (2002). Low Ci/Ca ratios over this period
(about 0.52) confirmed that highest photosynthetic activity
occurred in spring. The finding that the decreases in both
Ama and gsm from the second half of kernel-filling stage
onward occurred concomitantly with the increasing values
of Ci/Ca suggests that internal mesophyll limitations domi-
nated over stomatal limitations (Jifon and Syvertsen 2003).
The decreasing trend in both Am and Ap (Figure 7A–D)
during the stage of fruit dry mass accumulation was consis-
tent with the results reported by Walcroft et al. (2002) for
peach trees. However, this pattern is not a general charac-
teristic of fruit-bearing trees. Several studies have reported a
positive effect of fruit demand for assimilates on the
TREE PHYSIOLOGY ONLINE at http://www.treephys.oxfordjournals.org
385
photosynthetic rates of peach (DeJong 1986) and apple
(Palmer et al. 1997) trees.
There was a steep reduction in Amw (Figure 7B) from the
second half of kernel-filling stage onward that was indepen-
dent of irrigation regime and leaf class. However, after har-
vest, this declining trend slowed in the nf-leaves in the FI
treatment but was still marked in the DI treatment. Because
translocation of assimilates to perennial tree organs occurs
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post-harvest, the decrease in Am in DI trees during this
stage indicates that DI trees could be subjected to carbohy-
drate shortage in the following growth season (Esparza
et al. 2001a). This decrease probably explains the lower
Ama in DI trees than in FI trees, as well as their smaller
trunk diameter at the beginning of the growing season
(see section Materials and methods).
The lower Am observed in DI trees compared with FI
trees, along with the sustained decrease in Am observed in
trees in both treatments after Stages II and III, should be
analyzed to determine to what extent gs (Figure 7C) deter-
mines this pattern by restricting CO2 availability in the
mesophyll. Comparison of the seasonal trends in area-
based Ap (Figure 7D) and Am (Figure 7A) indicated a
rather similar behavior. Maximum Ap occurred in the
spring but declined during the remainder of the growing
season, independently of irrigation treatment. These results
provided indirect evidence that, in both irrigation treat-
ments, gs was not the main determinant of the decrease in
photosynthetic capacity (Jifon and Syvertsen 2003)
observed in our almond trees.
A negative correlation was observed between Am (area-
and dry-mass-based values) and Wa in both classes of
leaves, which seemed to be independent of irrigation regime
(Figure 8A and B). Higher values of Am in FI trees than in
DI trees could be related to differences in Wa (Figure 4A),
i.e., to differences in leaf thickness (Wa = leaf thick-
ness · leaf density). Thicker leaves are likely to have more
chloroplasts per unit of leaf area (Rieger and Duemmel
1992, Evans and Porter 2001) and therefore higher photo-
synthetic capacity.
Our results demonstrated that maximum assimilation
varied considerably over time with leaf age (Wilson et al.
2000), and that this seasonal effect dominated over other
factors such as leaf class and soil water availability. This
finding is supported by the data showing that Nw, which
is often used to describe the effects of leaf age (Field and
Mooney 1983, Reich et al. 1991), explained between 70%
and 92% of the seasonal variability in both Ama and Amw
(Figure 9A and B).
The marked decline of PNUEm throughout the growth
season (Figure 10A), irrespective of the irrigation treat-
ment, and the concomitant decrease in Ap (Figure 7D)
together suggest the seasonal dependence of N allocation
within the leaf. It is known that the allocation patterns of
chlorophyll and Rubisco can vary with leaf ontogeny
(Porter and Evans 1998, Rey and Jarvis 1998, Wilson
et al. 2000). The finding that seasonal changes in Ama and
386 NORTES, GONZALEZ-REAL, EGEA AND BAILLE
Na were poorly correlated, whereas both area- and mass-
based values of Am and leaf N concentration were strongly
related (Figure 9A and B), could be ascribed to changes in
the attribution of N within the leaf (Kull et al. 1998,
Dungan et al. 2003, Yasumura et al. 2006). The finding that
PNUEm was highly negatively correlated to Wa (Figure
10C) tends to support the hypothesis of a seasonal decrease
in mesophyll conductance.
The tight positive correlations between PNUEm and Nw
(Figure 10B), and between PNUEm and Wa (Figure 10C)
suggest that both leaf classes had an adequate N concentra-
tion and were functionally developed (Reich et al. 1991) in
both irrigation treatments. Because non-photosynthetic N
(Nwo) was lower in f-leaves, PNUEm was significantly
higher in f-leaves than in nf-leaves (Figure 10A) even
though there were no significant differences between leaf
classes in the irrigation regimes. However, when PNUEm
was calculated with respect to photosynthetic N (Nwp), no
significant effect of leaf class or irrigation regime was
observed. We found that the fraction of photosynthetic N
(fpN) decreased over the growing season (Figure 11), which
might support the hypothesis advanced by Wilson et al.
(2000) of a seasonally dependent fractional allocation of
leaf N to Rubisco.
Effect of soil water status imposed by the irrigation strategy
Lower Ama and Apa in nf-leaves of DI trees (17% and 15%,
respectively) than in FI trees prevailed throughout the
spring (Stages II and III) despite several heavy rainfall
(Figure 2) that maintained soil water near field capacity
in both irrigation treatments. This pattern, along with the
lower crown leaf area of DI trees compared with FI trees
(see section Materials and methods), suggested a carryover
effect of DI on whole-tree potential for CO2 uptake and
resource acquisition. Thus, after 4 years of sustained DI,
the DI trees appear to have acclimated to water stress but
at the cost of a lower photosynthetic capacity at the start
of the active vegetative growth stage.
We found that the photosynthetic capacity of almond
leaves was unaffected by a  100% decline in predawn leaf
water potential, likely indicating acclimation to the sus-
tained conditions of mild water stress (minimum value of
Wpd about 1 MPa) imposed during the previous 4 years.
The mild conditions of soil water deficit imposed on the
DI trees were not responsible for the seasonal reduction
in photosynthetic capacity. Although the interaction
between leaf class and irrigation regime was limited, with
both leaf classes exhibiting rather similar patterns of Am
and Nw in both irrigation regimes, the nf-leaves were more
responsive to soil water deficit than the f-leaves, with the
responses being mediated through leaf structural changes
(see Figure 6).
We conclude that, under the DI regime we applied, leaf
structural adjustments appeared to operate to maintain a
similar allocation of N resources in the leaves of fruit-bearing
TREE PHYSIOLOGY VOLUME 29, 2009
shoots, but to the detriment of N resources allocated to veg-
etative shoots. Foliar structural changes, along with changes
in nitrogen partitioning within the leaf – which might be
related to concomitant decreases in photosynthetic leaf N
concentration and the fractional allocation of leaf N to
Rubisco – were probably responsible for the decrease in pho-
tosynthetic capacity throughout the growth cycle.
Downloaded from https://academic.oup.com/treephys/article/29/3/375/1694688 by Universidade de Evora user on 09 March 2023
Acknowledgments
This study was supported by the Spanish Ministry of Science and
Technology (MCYT, grant AGL2002-04048-C03-02) and Euro-
pean Commission (IRRIQUAL project, FP6-2004-FOOD-3B-
C023120). Gregorio Egea was granted a research fellowship from
the FPU programme of the Spanish Ministry of Education.
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