Field Crops Research 58 (1998) 97107

Validity of various physiological traits as screening

criteria for salt tolerance in barley
R. Isla, R. Aragues*, A. Royo
Unidad de Suelos y Riegos, Servicio de Investigacion Agroalimentaria (D.G.A.), Laboratorio Asociado de Agronoma y Medio Ambiente
(DGA-CSIC), Apdo. 727, 50080 Zaragoza, Spain
Received 29 October 1997; received in revised form 25 December 1997; accepted 28 February 1998

Abstract

Limited knowledge of the physiological basis of the detrimental effects of soil salinity on growth and yield of barley and the
consequent lack of suitable screening traits are two reasons for the limited success of plant breeding in saline environments.
We assessed the relationships between grain yield, carbon isotope discrimination (
), canopy temperature, stomatal
conductance, and grain ash content in a set of barley cultivars grown in a soil salinity gradient imposed by a triple-line-source
sprinkler system. A linear increase in soil salinity produced signicant (P<0.05) and linear decreases in grain yield and
, and
increases in midday differential canopy temperature (T). For unit increase in soil solution electrical conductivity (ECss),

decreased by 0.2% and T increased by 0.38C. Increasing soil salinity from the control (non-saline) to a highly saline level
(ECss22 dS m1) decreased leaf stomatal conductance by 65% and increased grain ash content by 1129%. Grain yield and
grain
of 34 barley cultivars were signicantly (r0.58; P<0.01) correlated in non-saline conditions, suggesting that
is a
useful indicator of yield potential in barley. However, there was no signicant correlation between grain yield and grain

under highly saline conditions (r0.29; P>0.05). A multiple linear regression of
, stomatal conductance and grain ash
content on grain yield was highly signicant (R20.77, P<0.001) in non-saline conditions, but not in highly saline conditions.
We concluded that none of the studied characters would be useful in screening for high yield under saline environments, and
that grain yield under salt stress remains the only reliable means of identifying higher salt tolerance in barley. # 1998 Elsevier
Science B.V. All rights reserved.

Keywords: Ash content; Barley; Canopy temperature; Carbon isotope discrimination; Grain yield; Salt tolerance; Stomatal conductance

1. Introduction is a good target crop for breeding programs in such

Soil salinity is one of the principal abiotic factors
affecting crop yields in arid and semi-arid irrigated
areas (Szabolcs, 1989). In temperate climates, barley
*Corresponding author: Fax: 34 976 575501; e- mail:
raragues@syrsig.mizar.csic.es
salt-affected areas because of its inherently high sali-
nity tolerance (Maas and Hoffman, 1977) and, there-
fore, may offer a means for improving the productivity
of these environments. Since the classical selection of
materials based on their yield performance under
saline conditions has been largely unsuccessful, par-
tially due to the high variability of natural saline soils
(Richards, 1983), several authors (Noble and Rogers,

0378-4290/98/$19.00 # 1998 Elsevier Science B.V. All rights reserved.

PII S0378-4290(98)00088-4
98 R. Isla et al. / Field Crops Research 58 (1998) 97107
1992; Flowers and Yeo, 1995) have suggested the use
of physiological traits as alternatives to screening for
yield.
Richards (1992) found that, of the various crop
species evaluated in saline conditions, barley was
the most water-use efcient crop, presumably due
to its higher leaf area and faster leaf area growth,
as indicated also from the work of Lopez-Castaneda
and Richards (1994) in rainfed conditions. These
authors concluded that an increase in the water-use
efciency (WUE: the total aboveground dry matter
over total water transpired per unit of ground surface)
of crops grown in saline soils that are not frequently
irrigated could increase their productivity in these
conditions.
Farquhar and Richards (1984) showed that carbon
isotope discrimination (
) is linearly related to the
ratio (pi/pa) of the intercellular (pi) and atmospheric
(pa) partial pressures of CO2 in C3 plants. The pi/pa
ratio is determined by leaf stomatal conductance and
photosynthetic capacity and, therefore, by genetic and
environmental factors. Low
has been proposed as an
indicator of high WUE in C3 plants, since a negative
correlation between WUE and
has been reported for
wheat (Farquhar and Richards, 1984; Condon et al.,
1990; Ehdaie and Waines, 1993), rice (Dingkuhn
et al., 1991), and various other crops (Knight et al.,
1994).
However, the relationship between
and yield in
barley, and hence its utility as an indicator of yield, has
often been found to be positive, indicating that high
yield is associated with low WUE. Febrero et al.
(1994) and Romagosa and Araus (1991) found a
positive correlation between grain
and grain yield
among a set of barley cultivars. Austin et al. (1990)
and Craufurd et al. (1991) also found signicant
positive correlations between grain
and grain yield,
and that the correlations were strongest under water
stress conditions. These authors found a signicant
negative correlation between
and days from sowing
to heading, and between grain yield and days from
sowing to heading. They suggested that the grain

grain yield correlation was largely a consequence of
the genotypic variation in heading date, the earlier
cultivars (with high
and high yield) having an
advantage under terminal water stress conditions rela-
tive to the later ones (with low
). All the above
results imply a negative correlation between grain
yield and WUE, the opposite of what was suggested
by Richards (1992).
Pakniyat et al. (1997) reported that some barley
mutants grown in hydroponic culture had simulta-
neously a higher
, a lower Na shoot content, and
that their shoot growth was less depressed under saline
conditions than their respective parental lines. If it can
be conrmed that
and salinity tolerance are corre-
lated, a rapid advance in breeding barley for increasing
salinity tolerance may be expected. Handley et al.
(1994) have reported that the genes from chromosome
4 control
in barley. As genetic variability in
could
be easily determined through molecular marker tech-
niques these may offer a more efcient alternative to
screening for
.
The passive accumulation of silica through the
xylem in barley grown under eld conditions may
be also related to WUE (Walker and Lance, 1991).
Masle et al. (1992) and Mayland et al. (1993) proposed
leaf ash content as a trait correlated with WUE since it
was positively correlated with
. For barley, Febrero
et al. (1994) and for wheat Araus and Nachit (1996)
suggested the combined use of
and ash content of
grain as indicators of grain yield in rain-fed condi-
tions. Although the physiological relationship
between ash content and WUE is not as consistent
as that between WUE and
, further research on this
trait could be of interest to cereal breeders due to its
low cost and simplicity of measurement.
Farquhar et al. (1989) concluded that low
is
generally associated with low stomatal conductance.
A decrease in stomatal conductance under salinity
stress has been reported for barley by Shen et al.
(1994) and Benes et al. (1996), but the contribution
of decreased stomatal conductance to the reduction of
assimilation and overall growth rates among cultivars
grown under saline conditions remains uncertain.
Despite the similarities between water and salt
stress in their effects on plant growth, few attempts
have been made to quantify the effects of salinity on

as well as its potential utility as a breeding character
aimed at increasing grain yield under saline condi-
tions. Brugnoli and Lauteri (1991) with cotton and
bean, and Johnson (1991) with Agropyron found that
the
of plants subjected to salt stress was less than
that in non-stress conditions. The results of Pakniyat
et al. (1997), who reported that some barley mutants
grown in a hydroponic culture showed simultaneously
 R. Isla et al. / Field Crops Research 58 (1998) 97107 99

a higher
, lower Na shoot content, and lower shoot

growth depression under saline conditions than their
respective parental lines are of interest, but need to be
veried under eld salinity conditions and with a
larger set of barley cultivars.
The objectives of the work reported here were (1) to
study the effects of soil salinity on carbon isotope
discrimination, stomatal conductance, canopy tem-
perature and grain ash content in a set of barley
cultivars grown in controlled eld conditions, and
(2) to seek relationships among these traits and salinity
tolerance of barley assessed by its grain yield.
Fig. 1. Averaged values of soil salinity (ECa, electromagnetic
sensor readings) and irrigation water salinity (ECiw) measured in
each saline treatment of the TL93 and TLS94 experiments. The
2. Materials and methods bars are one standard deviation of the mean. The `EC-saline
treatment' regression lines are also shown.
2.1. Experimental arrangement

A triple-line-source sprinkler (TLS) system, Table 1 shows the general characteristics of the eld
described in detail by Aragues et al. (1992), was trials. Before sowing of the 1993 and 1994 experi-
installed during the 1992/1993 and 1993/1994 barley ments the eld was fertilised with 75 kg ha1 of N,
growing seasons at the experimental eld station of the P2O5 and K2O each. At tillering, 66 kg ha1 of N was
Agronomic Research Service-Diputacion General de also added.
Aragon (SIA-DGA, Zaragoza, Spain) located in the
central part of the Ebro river basin (08490 W, 418440 N).
The soil is a Typic xerouvent with a siltyclayloam
Table 1
texture. Briey, the TLS system consists of three General characteristics of field trials in years 1993 and 1994
parallel sprinkler lines spaced 15 m apart, a distance
1993 1994
equivalent to the wetted radius of a sprinkler. Sprink-
lers along the laterals are spaced 4.5 m apart. A saline Number of barley cultivars 16 18
solution made up of NaCl and CaCl2 (1:1, w/w) is Sowing date 27 Nov. 1992 19 Nov. 1993
injected in the centre line while the outer lines dis- Harvest date 2 July 1993 22 June 1994
Plot size 0.8 m2 1.6 m2
charge fresh water (EC<2 dS m1). This arrangement rows: number 3 6
produces a linear salinity gradient between the centre lengthwidth (m) 1.260.21 1.260.21
and the two outer laterals while providing an even Number of saline treatments 10 10
distribution of water (measured irrigation unifor- Number of replicates 1 1
mity>90%). Before and after each saline irrigation, Plants established 22017 20625
(number m2)
a 3 min fresh water irrigation is applied through each Number of saline irrigations 31 28
of the three laterals to reduce foliar salt absorption and First saline irrigation 3 Feb. 1993 31 Jan. 1994
injury. Last saline irrigation 31 May 1993 31 May 1994
For each barley cultivar, one strip with ten indivi- Seasonal saline irrigation 399 351
dual saline treatment areas (T9-highest salinity to T0- (mm)
Seasonal rain 128 118
lowest salinity) was designated between each lateral non-saline irrigation (mm)
pair. Pluviometers were installed in each treatment EC irrig. water a 3.2 to 19.9 1.9 to 17.3
area for measuring the volume and the EC of the (dS m1) T0 to T9
applied water during each irrigation. Fig. 1 shows that Seasonal evapotranspiration 241 271
both in 1993 and 1994 the average irrigation water (mm)
salinity (ECiw) gradients were linear (P<0.001). a
Average EC of all saline irrigations.
100 R. Isla et al. / Field Crops Research 58 (1998) 97107
Sixteen barley cultivars were sown in 1993: Alba-
cete, Barbarrosa, Begona, Mogador, Igri, Reinette,
Martin, Tunis, Pen, CA-54, Dpche-18, KVL-468,
Mut-4210, RPB7078, Atlas-76 and CM67. Eighteen
cultivars were sown in 1994: Albacete, Alpha, Bar-
barrosa, Cameo, Criter, Dpche-18, Igri, Kvl-468,
Malta, Mogador, Pen, Reinette, Acsad-60, Acsad-
176, Aglou, Annoceur, Asni and Merzaga. These
materials are commercial cultivars and breeding lines
from Spain and Morocco. Since we were seeking
relationships among characters rather than statistical
differences among cultivars, there were no replication
for a given salinitycultivar combination.
2.2. Soil water content and soil salinity
Soil samples were taken at various depths periodi-
cally in 1993 and 1994 in the control (T0), medium
(T4) and high salinity (T8) treatments and the gravi-
metric soil water content was measured in the labora-
tory. The values were fairly constant throughout the
growing season, close to eld capacity and without
statistical differences among the three salinity treat-
ments (data not given).
Soil salinity (apparent electrical conductivity, ECa)
in each plot (i.e., the saline treatments of each barley
cultivar) was periodically measured using a portable
electromagnetic sensor (EM-38, Geonics, Ont.,
Canada) placed on the ground in its horizontal dipole
position. In this conguration the EM-38 integrates
the salinity from the soil surface to a depth of approxi-
mately 1 m. Fig. 1 shows the ECa values measured in
1993 and 1994 in each saline treatment. These ECa
values are time-averages of seven (in 1993) and six (in
1994) sampling dates, and space-averages of 16 (in
1993) and 18 (in 1994) strips. A linear (P<0.001) soil
salinity gradient was obtained in both years. The mean
CVs (%) of the time-averaged ECa readings were 19%
in 1993 and 13% in 1994 and the mean CVs of the
space-averaged ECa readings were 10% in 1993 and
9% in 1994. These results indicate that the temporal
and, in particular, the spatial variability of soil salinity
in each saline treatment were low and would enable
valid comparisons to be made among the barley
cultivars.
Soil samples were taken periodically in the 1993
and 1994 eld trials for calibration of the EM-38
sensor and conversion of ECa to ECe (soil saturation
extract) through linear regression. From the pooled
data of 1993 and 1994 the calibration curve obtained
was
ECe050 cm 8:31 ECa 4:21 n 35; R2 0:75
Based on these ECe estimates, the measured
050 cm soil saturation percentage (mean55.5%)
and the measured 050 cm gravimetric soil water con-
tent (mean of 21.9% in 1993 and 20.8% in 1994), we
estimated the soil solution EC (ECss) to be 8.2 and
10.2 dS m1 (control treatment T0, in 1993 and 1994,
respectively), 14.2 and 16.5 (intermediate saline treat-
ment T4, in 1993 and 1994), and 23.1 and 22.7 (high
saline treatment T8, in 1993 and 1994). These values
indicate that the soil solution salinity gradients were
appropriate and reasonably similar in both years.
2.3. Carbon isotope discrimination

The effect of a continuous and linear soil salinity
gradient on
of leaves and grain was analysed by
sampling in each salinity treatment 20 ag leaves of
cvs. Albacete and Igri in 1993 and the grain from 20
ears of cv. Alpha in 1994. The effect of soil salinity on
the grain
of each barley cultivar was analysed by
sampling at maturity 20 ears in the control (T0) and
the high (T9) saline treatments in 1993, and in the
control (T0), medium (T4) and the high (T8) saline
treatments in 1994. The samples were oven dried at
608C, ground to a ne powder, and the carbon isotope
ratios (13 C=12 C) were determined in duplicate samples
by mass spectrometry (Isotope Services, Los Alamos,
NM, USA). The d13 C (p, relative to Pee Dee Belem-
nite, the international standard) values were converted
into carbon isotope discrimination (
, expressed as
%) as described by Farquhar et al. (1989), assuming
that the d13 C for atmospheric CO2 was 7.85%
relative to PDB.
2.4. Infrared thermometry
Canopy temperatures at the tillering (late March)
and booting (early May) stages were measured on 12
barley cultivars grown in 1994 in three salinity treat-
ments (T0, T4 and T8) and in cv. Alpha at the booting
stage in 1994 in ten salinity treatments (T0 to T9).
Replicated measurements were made with a Raynger
II-PAG model infrared thermometer (Rayteck Santa
 R. Isla et al. / Field Crops Research 58 (1998) 97107
Cruz, CA) by holding the thermometer about 0.7 m
above the canopy and aiming it to the south at an angle
of about 208 from the horizontal. Under these viewing
conditions a canopy area of 0.2 m in diameter com-
pletely lled the instruments eld of view. Canopy
temperatures were expressed as differences from the
air temperatures (T: canopy temperature minus air
temperature).
2.5. Stomatal conductance
Midday abaxial stomatal conductance measure-
ments were taken with a portable porometer LI-
1600 (Licor, Inc. Nebraska, USA) on 29 April 1994
on the ag leaves of twelve barley cultivars. The
measurements were made in triplicate in the control
(T0) and high (T8) salinity treatments.
2.6. Grain ash content
Duplicate samples of grain were taken in 1993 and
1994 from all the barley cultivars in the control (T0)
and high salinity (T8) treatments. The grain samples
were milled and a subsample of 3.0 g was oven-dried
at 1058C for 24 h, burnt in a furnace at 5508C for 8 h,
and the ash residue weighed. The ash content was
expressed as a percentage of kernel dry weight.
2.7. Grain yield
At the end of the 1993 and 1994 growing seasons
(Table 1), the plants in each plot were harvested and
Fig. 2. Effect of soil salinity (ECa) on grain yield of barely cultivars in 1993 (n16) and 1994 (n18). The regression lines and the
coefficients of determination are also shown.
101
the grain threshed out. Yields of grain were expressed
in g m2.
2.8. Statistical analysis
All the statistical analysis were performed using
SAS (SAS Inc., 1988) statistical software. Compari-
sons of means were performed using the t-test. Com-
parisons of slopes were made using a F-test, taking the
root mean error (RME) of the overall regression (all
cultivars) as the error for all pairwise comparisons.
When the data of the two years were pooled, all
variables were normalised to have a mean of zero
and a standard deviation of one for each year. The
selection of the variables for the multiple linear
regressions was performed using the stepwise method
with the normalised variables.
3. Results and discussion
3.1. Salinity effects on grain yield and carbon
isotope discrimination
In both 1993 and 1994, grain yield declined
linearly with increasing soil salinity (P<0.001;
Fig. 2). Grain yields were higher (especially at low
salinities), decreased more sharply with salinity, and
tted the linear model better in 1994 than in 1993.
This was probably because most of the cultivars
tested in 1994 were commercial cultivars, whereas
ve of those tested in 1993 were breeding lines
102 R. Isla et al. / Field Crops Research 58 (1998) 97107

Fig. 3. Effect of soil salinity (ECa) on carbon isotope discrimination (
) (a,b) and grain yield (c,d) of three barely cultivars. For each cultivar,
the regression lines of
and grain yield on soil salinity are also shown. Samples were from flag leaf in 1993 and from grain in 1994.

with very low grain yields under low or non-saline
conditions.
The linear soil salinity gradients imposed by the
TLS system produced a linear and signicant
(P<0.001) decrease in
of Albacete, Igri and Alpha
(Fig. 3(a) and (b)). These results are in agreement with
those of Pakniyat et al. (1997) with barley, those of
Brugnoli and Lauteri (1991) with cotton and beans and
those of Johnson (1991) with Agropyron, who also
found a decrease of
with salinity. The maximum
values of
were around 18% for the least saline
treatments for both the ag leaves (1993) and the grain
(1994), and the minimum values were around 15.5%
for the ag leaves (1993) and 13.5% in the grain
(1994) for the most saline treatments. Grain
mea-
sured in the saline treatments were lower than those
found by Austin et al. (1990) and Febrero et al. (1994)
for barley grown under rainfed conditions. These
differences may be attributed to differences in vapour
pressure decit and the continuous saline stress (from
early tillering to maturity) imposed in our experiments
in contrast to the predominantly terminal water stress
imposed in the experiments of the above-cited authors.
The two cultivars tested in 1993 behaved similarly
(Fig. 3(a) and (c)) in
and grain yield responses to
soil salinity (slopes not signicantly different,
P>0.05), although leaf
of Igri showed an initial
plateau and a threshold salinity of about
ECa1.4 dS m1. Leaf
of Albacete and grain

of Alpha decreased from ECa values of around
0.9 dS m1. Because some environmental conditions
were different and different tissues were sampled
in both years, the comparison between the 1993 and
1994 cultivars is not appropriate. The results of
1994 (Fig. 3(b) and (d)) reveal a parallelism between
the effect of soil salinity on grain yield and on
grain
.
Table 2 shows the mean  standard deviation, the
coefcient of variation (CV) and the minimum and
maximum values of grain
measured in barley
cultivars grown in two (1993) and three (1994) saline
treatments. Although the cultivars were not the same
in the two years, the
values in both the control (T0)
and in the high saline (T8) treatments were similar in
1993 and 1994. Based on these values and the soil
solution salinity intervals between T0 and T8 (ECss in
Table 2), we calculated that
decreased by 0.17% (in
1993) and 0.22% (in 1994) per unit increase in ECss.
In 1994,
decreased by 0.21% per unit increase in
ECss both between T0 and T4 and T4 and T8, con-
 R. Isla et al. / Field Crops Research 58 (1998) 97107 103

Table 2
Carbon isotope discrimination (
): Mean  standard deviation (SD), coefficient of variation (CV) and minimum and maximum values
measured on the grain from the ears of n barley cultivars grown in two (1993) and three (1994) saline treatments

Year Saline treatment n ECssa(dS m1)
%

meanstd min max CV (%)

1993 C (T0) 16 8.3 18.20.65 17.0 19.7 3.6
H (T9) 16 23.1 15.70.57 15.1 16.8 3.7
C (T0) 18 10.2 18.10.71 16.2 19.1 3.9
1994 M (T4) 12 16.5 16.80.48 15.9 17.6 2.8
H (T8) 18 22.7 15.40.58 14.4 16.4 3.8
a
Mean soil solution EC estimated from ECa readings, the ECeECa calibration curve and the measured mean gravimetric soil water content.

rming that the
-salinity response functions were trials where water stress was absent, differ from those
linear as shown in Fig. 3. obtained under terminal water stress by Austin et al.
The variability of
among cultivars was low as (1990) and Craufurd et al. (1991), who found that
indicated by the CV values of the cultivar means, earlier cultivars had higher
because their growth
which were in all cases less than 4%. However, the occurred when the vapour pressure decit was less
range between extreme cultivars was similar to that than during the corresponding growth stages of the
found in many other studies. The differences between later ones.
the maximum and minimum
values were close to
3% in T0, 1.7% in T4 and 1.7 to 2.0 in T8. Since 3.2. Canopy temperature and stomatal conductance
Farquhar and Richards (1984) concluded that each
unit decrease in
implies a 15% increase in WUE, Linear increases in soil salinity induced linear
these differences correspond to differences in WUE of increases in midday differential canopy temperatures
around 30%. (T) measured in cv. Alpha at its booting stage
The rankings (Spearman correlation) of the eight (Fig. 4). The differences in T between the lowest
cultivars common to 1993 and 1994 for their
values and highest salinity treatments were close to 58C,
were similar (rs0.75 in T0, and rs0.76 in T8, both indicating that the imposed salinity stress increased
signicant at P<0.05) suggesting that there were stomatal closure and induced large rises in leaf tem-
consistent differences among these cultivars in
. perature. Based on the regression equation shown in
Similar results were obtained by Condon et al.
(1987) with wheat and Handley et al. (1994) with
barley.
Averaging over 1993 and 1994, the
values of the
cultivars in T0 were signicantly correlated with those
in T8 (P<0.05), indicating that there is little culti-
varsalinity interaction. This suggests, in agreement
with the conclusion of Pakniyat et al. (1997), that
selection for
could best be performed in non-stress
conditions, where its range of variation is higher.
Date of owering recorded for all the cultivars
grown in T0 and T8 was not signicantly correlated
(P>0.05) with
. Also, no differences were found in

among the earliest and the latest cultivars (data not
Fig. 4. Effect of soil salinity (ECa) on differential canopy
given). These results suggest that the date of owering temperature (T: canopy temperatures minus air temperature) of
was, at most, only a minor factor inuencing
in our the cultivar Alpha measured at the booting stage. The regression
experimental conditions. Our results, obtained in eld line and the coefficient of determination are also shown.
104 R. Isla et al. / Field Crops Research 58 (1998) 97107
Table 3
Differential canopy temperature (T), stomatal conductance, and grain ash content (meanSD) obtained at the control (T0), intermediate (T4),
and high saline treatment (T8) for the two field experiments. The number of cultivars (n) is also presented
Character Date
T (8C) March, 23 1994
T (8C) May, 5 1994
T (8C) May, 6 1994
Stomatal conductance (mmol m2 s1) April, 29 1994
Grain ash content (% of dry weight) 1993
Grain ash content (% of dry weight) 1994
Means with the same letter were not significantly different (P>0.05).
Fig. 4, the ECe-ECa calibration curve, and the mea-
sured mean gravimetric soil water content, each unit
increase in ECss led to an increase of 0.168C in T.
The average T of the 12 barley cultivars measured
on three sampling dates in 1994 increased with salinity
(T0, T4 and T8) (Table 3). The differences in canopy
temperatures, averaged over cultivars, between treat-
ments were clearly signicant (P<0.05) at tillering,
but not at booting. For these sampling dates, the
average increase in T per unit increase in ECss
was similar to that found in cv. Alpha (i.e., increases
between 0.13 and 0.218C per dS m1 increase in
ECss).
Differences in canopy temperature among the 12
cultivars increased with increases in soil salinity.
Thus, the average T range for the three sampling
dates was 4.08C in T0 and increased to 6.48C in T4 and
7.08C in T8. These differences in T could be partially
attributed to differences in phenology and senescence
among cultivars caused by soil salinity and by cultivar
differences in leaf colour.
Midday stomatal conductance was signicantly
reduced (P<0.05) by soil salinity (Table 3). The aver-
age stomatal conductance of the 12 cultivars measured
in the high salinity treatment was 65% lower than that
measured in the control. Similar results were obtained
with barley by Shen et al. (1994). Since the photo-
synthetic apparatus is quite tolerant to salinity (Robin-
son et al., 1983), the observed decline in assimilation
rate under salinity should mainly be attributed to the
decreases in stomatal conductance.
Stomatal conductance varied substantially among
cultivars, as indicated by the high CV of the means in
the control (CV32%) and, especially, in the high
salinity treatment (CV62%). This variability could
n Control (T0) Intermediate (T4) High (T8)
12 0.080.44 a 1.281.75 b 4.085.71 c
12 1.383.12 a 2.084.60 a 2.985.59 a
12 0.441.90 a 2.043.88 ab 2.944.64 b
12 92.329.9 a 32.920.5 b
16 2.680.24 a 2.960.64 a
18 2.240.43 a 2.900.40 b
potentially be exploited in screening, although stoma-
tal conductance are necessarily `spot' measurements
and can be substantially affected by micro-environ-
mental conditions.
3.3. Grain ash content
The average ash content of grain in 16 (1993) and
18 (1994) cultivars was signicantly (P<0.05) affected
by soil salinity in 1994, but not in 1993 (Table 3).
Average grain ash content measured in the high sali-
nity treatment was 11% (1993) and 29% (1994) higher
than that in the control treatment. These ash content
values were similar or slightly lower than those found
by Febrero et al. (1994) in non-saline conditions,
indicating that the grains do not accumulate salts in
substantial amounts, even though they were wetted by
sprinkling saline waters. The variability in ash content
among barley cultivars was relatively low, as shown by
the CV of the means, which were 15% in 1993 and
22% in 1994. This low variability limits the use of this
measurement as a tool for screening purposes.
3.4. Relationships among measured parameters
The correlation coefcients between
and grain
yield among cultivars for each year and salinity treat-
ment were not signicant (P> 0.05), with the excep-
tion of the control treatment in 1994 (P<0.01).
Averaging over 1993 and 1994, the correlation
between normalised
and normalised grain yield
was signicant (P<0.01) in the control treatment
(Fig. 5). This positive relationship between
and
grain yield found in non-stress conditions accords
with results of Romagosa and Araus (1991) with
 R. Isla et al. / Field Crops Research 58 (1998) 97107
Fig. 5. Relationships between pooled 1993 and 1994 normalised
grain yield and carbon isotope discrimination (
) in the control
(T0) and high saline (T8) treatments. The regression lines and the
correlation coefficients are also shown.
barley and Kirda et al. (1992) with durum wheat, and
shows that
may be a good indicator of yield
potential in barley. Since
and WUE are negatively
correlated (Farquhar and Richards, 1984; Knight et al.,
1994), our results suggest that grain yield and leaf
WUE are negatively correlated in non-stress condi-
tions, implying that a maximisation of yield occurs via
a maximisation of stomatal opening, despite the appar-
ent decrease in WUE. We consider that a major reason
of the lack of signicant relationships among yield and
the characters studied under the high saline treatment
may be the small genetic variability among the culti-
vars since the standard deviation of grain yield was
98 g m2 (16 cultivars) in 1993 and 69 g m2 (18
cultivars) in 1994, corresponding to coefcients of
variation of 43% and 25% respectively. This could be
one reason by which the correlation between normal-
ised
and normalised grain yield data of pooled 1993
Table 4
Summary of results of the stepwise multiple linear regression analysis of 1994 data obtained in the control (T0) and high saline (T8)
treatments. The model is: grain yieldf(
, stomatal conductance, grain ash content, average canopy temperature)
Independent Variable a Control (T0)
Parameter estimate
**

107.3
*
Stomatal conductance 85.2
*
Grain ash content 59.7
a
Only the variables with a significance of P<0.15 were included in the model. The variables were previously standardized.
Significance: NS, P>0.05; *, P< 0.05; **, P< 0.01.
105
and 1994 years for the high saline treatment, although
positive, was only signicant at P0.1 (Fig. 5).
The results obtained with 12 barley cultivars grown
in 1994 indicate that, for a given salinity treatment,
stomatal conductance and canopy temperature were
not signicantly correlated (P>0.05), and neither vari-
able was correlated with
. Likewise, no signicant
correlations were found between grain yield and
canopy temperature or stomatal conductance in any
saline treatment. These results do not agree with those
obtained under water stress conditions by Blum et al.
(1982) in wheat or by Garrity and O'Toole (1995) in
rice, who found signicant negative correlations
between canopy temperature and grain yield.
Pooled 1993 and 1994
and grain ash content
values for 34 barley cultivars were not signicantly
correlated (P>0.05), either in control or in saline
conditions. This result suggests that mineral accumu-
lation in kernels is not related to the integrated WUE.
A similar result was obtained by Febrero et al. (1994)
with barley under rainfed conditions. This lack of
correlation supports the view of these authors that
mineral accumulation in kernel occurs actively
through the phloem rather than passively through
the xylem.
Stepwise multiple linear regression (MLR) per-
formed for 12 cultivars grown in 1994 in non-saline
conditions (T0) between grain yield as the dependent
variable and
, stomatal conductance, grain ash con-
tent, and canopy temperature as the independent
variables indicated that
, stomatal conductance
and grain ash content partially explain the variability
in yield among these barley cultivars (Table 4). Based
on this analysis, grain yield will be positively and
signicantly correlated with
(at P<0.01) and sto-
matal conductance (at P<0.05), and negatively corre-
High saline (T8)
2
Partial R (%) Parameter estimate Partial R2
39 NS
21 NS
17 NS
106 R. Isla et al. / Field Crops Research 58 (1998) 97107
lated (at P<0.05) with grain ash content. The coef-
cient of determination of the multiple linear regression
was 0.77 (signicant at P<0.001), indicating that grain
yield could be estimated with high precision (standard
error of the estimation77 g m2) from the combina-
tion of these independent variables. However, it
should be emphasised that
and stomatal conduc-
tance are not really independent in the physiological
sense.
The similar MLR analysis performed for 12 barley
cultivars grown in 1994 in the high saline treatment
(T8) indicates that grain yield is not signicantly
correlated (P>0.05) with the independent variables
included in the analysis. These results are in general
accordance with those reported previously. Our con-
clusion is that selection of high yielding cultivars for
saline environments may not be performed by using
these traits as screening criteria.
4. Conclusions
A linear increase in soil salinity resulted in a linear
and signicant (P<0.05) decrease in grain yield and
carbon isotope discrimination (
) and a linear
increase in midday differential canopy temperatures
(T) in barley. For each unit increase in soil solution
EC,
decreased by 0.2% while T increased
by 0.38C.
Increasing soil salinity reduced leaf stomatal con-
ductance and increased grain ash content. Thus, the
average stomatal conductance of 12 barley cultivars
measured in the high salinity treatment was 65% lower
than that measured in the control, whereas the average
grain ash content measured in the high salinity treat-
ment was, depending on year, 11% to 29% higher than
that measured in the control.
Normalised grain yield and
of 34 barley cultivars
grown in non-saline conditions were signicantly
correlated (r0.58; P<0.01), indicating that
may
be a good indicator of yield potential, and suggesting
that grain yield and water-use efciency are negatively
related. However, under saline conditions, the correla-
tion was not signicant. The combination of
, sto-
matal conductance and grain ash content as
statistically independent variables regressed on grain
yield as the dependent variable gave a highly signi-
cant coefcient of determination (R20.77, P<0.001).
These traits could be used in screening of barley
cultivars for yield potential (i.e., in non-saline condi-
tions). On the other hand, the same multiple regression
performed for the barley cultivars grown under salinity
stress was not signicant.
Our results obtained under salinity stress conditions
are different to those reported by several authors for
barley grown under water stress conditions, where
signicant correlations were obtained among grain
yield and various of the characters studied. Therefore
barley behaves differently under these two stresses.
Finally, from our data, we were not able to identify one
or a combination of the characters studied which
would be useful in selecting high yielding barley
cultivars in saline environments.
Acknowledgements
This research was supported by a CICYT (Ministry
of education, Spain) research project founds and a
CONAI (Government of Aragon, Spain) fellowship to
R. Isla. Our thanks are due to J. Gaudo, M. Izquierdo,
T. Molina, and L. Naval for technical assistance and to
Dr. R. Austin for his critical review.
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