Environmental and Experimental Botany 54 (2005) 193201

Evaluation of some nutritional and biochemical indicators in

selecting salt-resistant tomato cultivars
Melchor Juan, Rosa M. Rivero, Luis Romero, Juan M. Ruiz
Department of Plant Physiology, Faculty of Sciences, University of Granada, 18071 Granada, Spain
Accepted 8 July 2004

Abstract
In the present work, in order to identify the most reliable nutritional and biochemical indicators for improving salt tolerance in
one of the most important horticultural crops worldwide tomato, 10 commercial cultivars were subjected to salinity stress. Here,
we show that the most salt-resistant tomato cultivars (cv. Brillante and cv. Jaguar) are characterized by reduced uptake and foliar
accumulation of Na+ and Cl , increased K+ uptake, and greater synthesis of sucrose, carotenoids, and thiol groups, with the
consequent reduction in lipid peroxidation and therefore, oxidative damage; all this translates as greater biomass production in
the two cultivars with respect to the other tomato cultivars examined. In addition, our results reveal the validity and effectiveness
of certain nutritional and biochemical indicators of salt stress, such as the K+ :Na+ ratio and sucrose, indicating the importance
of lipid peroxidation as the determinant physiological process in the selection of salinity-tolerant tomato plants.
2004 Elsevier B.V. All rights reserved.
Keywords: Lycopersicon esculentum L.; Plant indicators; Salinity stress; Salt tolerance

1. Introduction
The detrimental effects of salts on plants result not
only from a water deficit with relatively high solute
concentrations in the soil but also from specific Cl
Abbreviations: DMRT, Duncans multiple range test; DTNB,
5-5-dithiobis(2-nitrobenzoic acid); DTT, dithiothreitol; EDA,
ethylenediamine tetraacetic acid; GSH, glutathione; LOX, lipoxygenase; MDA, malondialdehyde; MES, morpholinoethanesulfonic
acid; ROS, reactive oxygen species; SH, sulfhydril
Corresponding author. Tel.: +349 58 243255;
fax: +349 58 248995.
E-mail address: jmrs@ugr.es (J.M. Ruiz).
0098-8472/$ see front matter 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.envexpbot.2004.07.004

and Na+ stress. The result is a wide variety of physiological and biochemical changes in plants that inhibit
growth and development, reduce photosynthesis, respiration, and protein synthesis, and disrupt nucleic acid
metabolism (Levine et al., 1990; Zhang and Blumwald,
2001; Sairam et al., 2002).
Corrections of the salinity problems are usually expensive and generally considered only temporary solutions. Selection and breeding of the cultivars that can
grow and provide economic yield under the saline conditions constitute more permanent and complementary
solutions to minimize the repercussions of the salinity
(Dasgan et al., 2002; Ashraf and McNeilly, 2004). A

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M. Juan et al. / Environmental and Experimental Botany 54 (2005) 193201

better understanding of the physiological and biochemical mechanisms conferring salinity tolerance is key for
developing selection and breeding strategies. However,
despite widespread research into the salinity tolerance
of plants, mainly on water relationships, photosynthesis, and accumulation of various inorganic ions and organic metabolites, little is known on the metabolic sites
at which salt stress damages plants and, conversely, the
adaptive mechanisms in plants to survive salinity stress
(Munns, 1993, Munns et al., 2002).
Currently, for a number of important agricultural
crops, no well-defined plant indicators for salinity tolerance are available to plant breeders to improve salt
tolerance. Most screening techniques to assess such tolerance in plants use proline accumulation as one of the
most important physiological parameters (Petrusa and
Winicov, 1997; Ali et al., 1999; Garg et al., 2002; Hsu
et al., 2003; Abraham et al., 2003; Ashraf and Harris, 2004). However, proline accumulation cannot be
regarded as a reliable marker for salt tolerance, as it
accumulates under various stress conditions such as
temperature and drought (Kuznetsov and Shevyakova,
1997; Ruiz et al., 2002; Ashraf and Harris, 2004). In
addition, under saline conditions, the accumulation of
other compounds such as sugars, which at higher temperatures are non-toxic to cytoplasmic functions, allowing turgor maintenance and/or protection of macromolecular structural against the destabilizing effect of
the decrease in water activity, have also been reported
in many species (Popp and Smirnoff, 1995; Ashraf and
Fatima, 1995; Murakeozy et al., 2003).
In addition to the study on the behavior of these
compatible compounds, the analysis of the K+ :Na+
ratio is a useful indicator of the degree of plant resistance to salinity. In general, salinity-sensitive plants undergo Na+ and K+ depletion (Cuartero and FernandezMunoz, 1999; Maathuis and Amtmann, 1999; PerezAlfocea et al., 1996; Sairam et al., 2002), lowering yield
and biomass productivity (Asch et al., 2000). Coupled
with this nutritional parameter, the activity and content
in photosynthetic pigments are also amply used in these
types of studies. Thus, lower chlorophyll contents due
to salinity have been reported and therefore, chlorophyll contents have been proposed as one of the attributes of salt tolerance in crops (Abd el Samad, 1993;
Hernandez et al., 1995; Romero et al., 1997; Sairam
et al., 2002). Also, reductions in carotenoid contents
under salinity stress have also been reported (Abd el

Samad, 1993; Romero et al., 1997). Carotenoids are

responsible for quenching singlet oxygen, and hence
their comparative levels in a cultivar may determine relative stress tolerance (Knox and Dodge, 1985; Sairam
et al., 2002).
It is now widely accepted that reactive oxygen
species (ROS) are responsible for diverse stressinduced damage to macromolecules and ultimately to
cell structure (Mittler, 2002; Vranova et al., 2002).
Some researchers have examined the generation of
ROS and lipid peroxidation in plants in relation to
salinity stress (Hernandez et al., 1993; Gossett et
al., 1994; Gueta-Dahan et al., 1997; Sairam et al.,
2002). Dionisio-Sese and Tobita (1998) demonstrated
increased lipid peroxidation and electrolyte leakage as
well as Na+ accumulation in the leaves of the saltsensitive rice varieties Hitomebore and IR28 under
salinity stress.
Given that tomato is a major crop worldwide, and
that its cultivation is concentrated in semi-arid regions,
where saline waters are frequently used for irrigation
(Santa-Cruz et al., 2002). In the present work, we study
the behavior of some of the nutritional and biochemical attributes, cited above in 10 tomato cultivars subjected to a concentration of 100 mM NaCl in order to
determine most reliable indicators for distinguishing
salt-sensitive from salt-tolerant plants.
2. Material and methods
2.1. Plant material and growth conditions
Ten cultivars of tomato plants (Lycopersicon esculentum L.) used in the present study were Cencara,
Tesoro, Sesenta, Marimba, Caramba, Zinac, Daniela,
Royesta, Brillante and Jaguar. Seeds of the 10 cultivars were germinated and grown for 30 days in cell
flats (cell size, 3 cm 3 cm 10 cm) filled with peatlite mixture, and the flats were placed on benches under the experimental greenhouse in southern Spain
(Granada, Sanivet S.L.). The 30-day-old seedlings
were transferred to a cultivation chamber under controlled environmental conditions with relative humidity
of 6080%, temperature 25 C/15 C (day/night) and
16 h/8 h photoperiod at a PPFD of 350 mol m2 s1
(measured at the top of the plants with a 190 SB
quantum sensor, LI-COR Inc., Lincoln, NE, USA).
The plants were grown in pots (25 cm upper diame-

M. Juan et al. / Environmental and Experimental Botany 54 (2005) 193201

ter, 17 cm lower diameter, 25 cm in height) each pot

containing one plant. Pots were filled with vermiculite. The plants were grown for 15 days (from day
30 until day 45 after sowing). Before the start of salt
treatment, the plants were grown in a nutrient solution and each received: 6 mM KNO3 , 2 mM NaH2 PO4 ,
1.25 mM CaCl2, 1.5 M MgSO4, 10 M Fe-EDDHA,
2 M MnSO4 , 1 M ZnSO4, 0.25 M CuSO4 , 0.1 M
(NH4 )6 Mo7 O24 and 25 M H3 BO3 . The nutrient solution (pH 5.56.0) was renewed after every 3 days and
the vermiculite partly rinsed with Millipore-filtered water in order to avoid nutrient accumulation.
Forty five days after sowing, NaCl treatment
(100 mM) was started and maintained for 21 days,
when most of the plants showed clear symptoms of
NaCl toxicity. For each tomato variety, the experimental design was randomized into a complete block with
three replicates of six plants each. The experiment was
completely repeated three times (n = 9) under the same
conditions as above.
2.2. Plant sampling
Leaves of these plants were sampled on day 66 after
sowing. Leaf samples were standardized by using only
fully expanded leaves from the middle part of plants
in each replicate, as these reflect most clearly, from
the nutritional and metabolic standpoint, the effects of
the treatment applied. About 20 leaves per plant were
sampled during the middle of the daylight photoperiod.
The material was rinsed three times in distilled water
after disinfection with non-ionic 1% detergent (Wolf,
1982), then blotted on filter paper. At each sampling,
fresh leaf matter was used fresh for the analysis of pigments (chlorophyll a, b and carotenoids), proline, sucrose, total phenols, lipid peroxidation (lipoxygenase
(LOX, EC 1.13.11.12) activity and malondialdehyde
(MDA) content) and total SH contents. Another subsample was dried in a force-air oven at 70 C for 24 h,
ground in a Wiley mill and then used for the analysis
of the nutrient concentration (Na+ , K+ and Cl ). Dry
weight was recorded and expressed as grams of dry
weight (DW) per leaf.
2.3. Biochemical determinations
To measure chlorophylls and carotenoids, the procedure described by Wellburn (1994) was followed. To

195

determinate the foliar area, leaf discs of 7 mm diameter and 0.125 g were collected. The chlorophylls and
carotenoid contents were expressed as microgram per
centimetre square, using Wellburn (1994) equations.
Free proline and sucrose were determined in 95%
ethanol extracts from leaves. Samples of 0.5 g of tissues freshly harvested were crushed in 5 ml 95% (v/v)
ethanol. The insoluble fraction of the extract was
washed twice with 5 ml of 70% ethanol. All soluble
fractions were centrifuged at 3500 g for 10 min.
The supernatants were collected and stored at 4 C
for proline and sucrose determination (Irigoyen et al.,
1992). The free proline content was measured according to the method described by Paquin and Lechasseur
(1979). Sucrose content was determined spectrophotometically at A650 , using the colorimetric assay with
anthrone reagent (Irigoyen et al., 1992).
Phenolics of plant material were extracted with
methanol. Total phenolic content was assayed quantitatively by A675 with FolinCiocalteau reagent
(Singlenton and Rossi, 1965).
For the assay of MDA, leaves were homogenized with 5 ml of 50 mM buffer solution, which
contained 0.07% of NaH2 PO4 2H2 O and 1.6%
Na2 HPO4 12H2 O, triturated with a mortar and pestle,
and centrifuged at 20,000 g for 25 min in a refrigerated centrifuged. The supernatant was used for the
determination of MDA content. For measurement of
MDA content, 4 ml of 20% trichloroacetic acid containing 0.5% thiobarbituric acid was added to a 1-ml
aliquot of the supernatant. The mixture was heated at
95 C for 30 min and then quickly cooled in an ice bath.
After the tube was centrifuged at 10,000 g for 10 min,
the absorbance of the supernatant was read at A532 . The
value for the nonspecific absorption at A600 was subtracted from the A532 reading. The concentration of
MDA was calculated using MDAs extinction coefficient of 155 mM1 cm1 (Heath and Packer, 1968; Fu
and Huang, 2001).
LOX activity was measured according to MinguezMosquera et al. (1993) using 50 mM K-phosphate
buffer (pH 6.0) for extraction. Neither the addition of
Triton X-100 to improve the solubility of the enzyme
nor the addition of dithiothreitol (DTT) to protect SHgroups from oxidation improved the assay results. The
reaction mixture consisted of 20 l crude extract and
0.5 mM linoleic acid in 50 mM K-phosphate buffer (pH
6.0). LOX activity was calculated following the rise in

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M. Juan et al. / Environmental and Experimental Botany 54 (2005) 193201

the extinction at 234 nm using an extinction coefficient

of 25,000 M1 cm1 (Axelrod et al., 1981; Egert and
Tevini, 2002). The protein content of the extract was
measured according to Bradford (1976) with BSA as
the standard. The activity was based on the protein concentration of the extract and expressed in units (U) per
milligram protein, where 1 U is defined as an amount
of enzyme the utilizes 1 mol linoleic acid min1 .
Total sulfhydryl (SH) content was measured accoding to De Kok et al. (1988). Leaf tissue was extracted in a mixture of 80 mM sulfosalicylic acid, 1 mM
ethylenediamine tetraacetic acid (EDTA) and 0.15%
sodium ascorbate (w/v). The homogenate was centrifuged at 30,000 g for 15 min. Total SH content
was determined by adding 1 ml of supernatant extract
to a mixture of 1 ml 50 mM morpholinoethanesulfonic
acid (MES), pH 5.8. After 10 min incubation at 30 C,
0.1 ml 5-5 -dithiobis(2-nitrobenzoic acid) (DTNB) (in
20 mM potassium phosphate, pH 7.0) and 1 ml 200 mM
TrisHCl (pH 8.0) were added and the yellow colour
developed, was measured at A415 .
2.4. Nutrient determinations
For the determination of the total amounts of Na+
and K+ , dry leaf (0.1 g) was subjected to sulfuric acid
digestion in the presence of H2 O2 (Wolf, 1982), and
diluted with distilled water. Total Na+ and K+ contents
were directly measured by flame spectrophotometry
(Lachica et al., 1973). Cl was analyzed after aqueous
extraction of 0.2 g of dried and ground leaf material
in 10 ml of distilled water. Cl content was measured
according Diatloff and Rengel (2001).
2.5. Statistical analysis
Analysis of variance was used to assess the significance of treatment means. Differences between plant
cultivars means were compared using the LSD and
Duncans multiple range test (DMRT). A correlation
analysis was also conducted to determine the relations
among different variables.

3. Results and discussion

One of the harmful effects of salinity on plant growth
involves the excessive accumulation of Na+ and Cl

in the leaves (Zhang and Blumwald, 2001; Munns et

al., 2002; Ashraf and Harris, 2004). This accumulation
under saline conditions depends on the plants capacity
to limit the uptake of these elements (Koval and Koval,
1996). In this regard, we can use the term excluders
for plants that block or reduce the uptake and accumulation of Na+ and Cl , and includers for plants that
take up large quantities of salts (Romero et al., 1997).
In the breeding programs, the selection of plants that
exclude these ions is essential for improving salinity
tolerance in a number of agricultural crops (Ashraf,
1994).
In tomato plants subjected to salinity, the genotype determines Na+ and Cl accumulation in leaves
(Table 1). For both Na+ and Cl the cultivars Brillante and Jaguar (excluders plants) presented the lowest foliar concentrations in these ions, while Royesta
(includers plant) registered the highest concentrations.
By studying these differences among cultivars, we formulated a valid experimental model to analyze the responses of different nutritional and biochemical indicators under saline conditions.
Plants growing under saline conditions accumulate more of Na+ , resulting in ionic imbalance, specific ion effects, and nutrient-deficiency symptoms in
plants. Decreased K+ , Ca2+ , Mg2+ uptake reportedly
depresses growth at higher Na+ concentrations (Sairam
et al., 2002). The control of Na+ accumulation and
high shoot K+ :Na+ ratios may enhance salt tolerance
in tomato crops (Perez-Alfocea et al., 1996; Cuartero
and Fernandez-Munoz, 1999; Al-Karaki, 2000; Dasgan
et al., 2002). In our experiment, the results for K+ and
the K+ :Na+ ratio were similar to those indicated by
other authors. Table 1 reflects that the cultivars capable
of accumulating lower foliar Na+ concentrations (cv.
Brillante and cv. Jaguar, Table 1) presented higher foliar
K+ concentrations and higher K+ :Na+ ratios (Table 1).
By contrast, cv. Royesta registered the highest foliar
Na+ concentrations.
The excess foliar accumulation of Na+ and Cl inhibits plant growth and development (Munns et al.,
2002; Ashraf and Harris, 2004). One of the mostly
widely used agricultural indices to define stress tolerance are data for plant biomass and yield (Sairam et al.,
2002). The effect of the lower foliar concentrations of
Na+ and Cl in the cultivars Brillante and Jaguar was
evident in the leaf biomass, as these cultivars presented
the highest production values (Table 1). On the con-

M. Juan et al. / Environmental and Experimental Botany 54 (2005) 193201

197

Table 1
Concentration of Cl , total Na+ , total K+ , K+ /Na+ ratio and foliar biomass in 10 commercial cultivars of tomato plants subjected to salinity
stress (100 mM NaCl)
Cultivar

Cl

Total Na+

Total K+

K+ /Na+ ratio

Foliar biomass

Cencara
Tesoro
Sesenta
Marimba
Caramba
Zinac
Daniela
Royesta
Brillante
Jaguar
LSD0.05
Significanceb

10.36 ba
10.91 b
10.62 b
10.94 b
10.79 b
11.08 b
10.12 b
14.77 a
7.73 c
8.21 c
2.14
***

7.32 b
7.55 b
7.18 bc
7.73 b
7.01 bc
7.46 b
7.55 b
13.19 a
5.22 d
6.55 c
0.70
***

5.11e
4.74 e
4.83 e
5.48 e
6.12 cd
6.53 c
5.22 e
5.65 de
9.32 a
7.93 b
0.72
***

0.70 cd
0.63 cd
0.67 cd
0.71 c
0.87 c
0.87 c
0.69 cd
0.43 d
1.78 a
1.21 b
0.27
***

0.012 b
0.014 b
0.012 b
0.015 b
0.015 b
0.012 b
0.013 b
0.006 c
0.021 a
0.018 ab
0.003
**

Foliar concentration of Cl , total Na+ and K+ expressed in mg g1 DW. Foliar biomass expressed as g DW/leaf.
a Values followed by the same letters within each column are not different using the Duncans multiple range test at 95%.
b Levels of significance are represented by (*) at P < 0.05, (**) at P < 0.01, (***) at P < 0.001 and NS: not significant.

trary, the lowest production of leaf biomass was found

in Royesta, defining it as the most salt-sensitive of all
the cultivars studied (Table 1). The regression analysis
between the foliar concentrations of Na+ and Cl and
foliar biomass confirmed the phytotoxicity of excess
accumulation in these ions (Na+ concentrationfoliar
biomass, r = 0.83 ***; Cl concentrationfoliar
biomass, r = 0.84 ***), clearly defining the cultivars
most tolerant to this type of stress (cv. Brillante and cv.
Jaguar) as well as the most sensitive (cv. Royesta).
As indicated in the Section 1, the K+ :Na+ ratio
has been used as a nutritional indicator by a number
of authors to select salt-tolerant plants, given the directly proportional relationship of this attribute with
biomass production (Perez-Alfocea et al., 1996; Cuartero and Fernandez-Munoz, 1999; Maathius and Amtmann, 1999; Asch et al., 2000; Sairam et al., 2002).
In the present work, we define the K+ :Na+ as reliable
indicator of salt tolerance, in view of its close and significant relationship with foliar biomass production in
tomato (r = 0.82 ***).
Decreased pigment contents due to salinity have previously been reported, with chlorophyll and carotenoid
contents being suggested as one of the parameters of salt tolerance in crop plants (Hernandez
et al., 1995; Romero et al., 1997; Srivastava and
Sharma, 1998; Sairam et al., 2002), prompting their
wide use in screening programs. The lower foliar
Na+ and Cl concentrations in the cultivars Brillante and Jaguar (Table 1) did not result in the

Table 2
Foliar concentration of pigments (microgram per centimetre square)
in 10 commercial cultivars of tomato plants subjected to salinity
stress (100 mM NaCl)
Cutivar

Chl a

Chl b

Carotenoids

Cencara
Tesoro
Sesenta
Marimba
Caramba
Zinac
Daniela
Royesta
Brillante
Jaguar
LSD0.05
Significanceb

8.14 ba
7.58 b
9.45 a
6.39 c
8.88 ab
10.19 a
6.56 c
5.96 c
8.97 ab
7.19 b
0.83
***

6.77 bc
4.83 d
6.99 b
5.47 cd
7.49 b
8.45 a
5.52 cd
5.16 d
5.99 c
6.16 c
0.72
**

3.31 bc
3.01 c
3.56 b
2.89 c
3.82 b
4.01 b
3.70 b
2.44 c
5.22 a
5.01 a
0.60
***

a Values followed by the same letters within each column are not
different using Duncans multiple range test at 95%.
b Levels of significance are represented by (*) at P < 0.05, (**) at
P < 0.01, (***) at P < 0.001 and NS: not significant.

expected increase in Chl a and Chl b contents

(Table 2), reflecting the weak relationship found between these parameters (Na+ concentrationChl a, r
= 0.53 NS; Na+ concentrationChl b, r = 0.31
NS; Cl concentrationChl a, r = 0.33 NS; Cl
concentrationChl b, r = 0.15 NS). These data suggest that, under our experimental conditions in the
tomato cultivars tested, Chl a and Chl b would not be
good indicators for salt stress tolerance, as reflected by
the simple-regression analysis between these pigments

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M. Juan et al. / Environmental and Experimental Botany 54 (2005) 193201

and foliar biomass production (Chl afoliar biomass, r

= 0.20 NS; Chl bfoliar biomass, r = 0.016 NS).
On the contrary, Table 2 shows that the highest
levels of carotenoids corresponded to the cultivars
Brillante and Jaguar, and the lowest to cv. Royesta,
presenting an inverse relationship between this pigment and the foliar concentrations of Na+ and Cl
(Na+ concentrationcarotenoids, r = 0.78 **; Cl
concentrationcarotenoids, r = 0.86 ***). According
to these results, the analysis of carotenoids could be
used as a simple and effective method to select salttolerant tomato cultivars, since the relationship with
foliar biomass production was significant (r = 0.756
**).
Among the compounds considered compatible, perhaps the most thoroughly studied in relation to the
response to saline stress in plants is proline. Petrusa
and Winicov (1997) found that salt-tolerant alfalfa
plants rapidly doubled their proline content in the roots,
whereas in salt-sensitive plants the increase was slow.
However, the role of the proline in osmoregulation and
salt tolerance has generally been questioned. For example, higher proline accumulation was found in salttolerant Brassica juncea plants with better growth than
the control (Kirti et al., 1991). Also, in relatively salttolerant plants of B. juncea showed a higher degree of
osmotic adjustment in the leaves and a higher critical
point concentration on NaCl, at which the endogenous
level of free proline rose sharply, than did the relatively
salt-sensitive genotypes (Jain et al., 1991). In contrast,
Tal et al. (1979) reported more proline accumulation
in salt-sensitive species of tomato than in tolerant wild
relatives. Aziz et al. (1998) reported a negative relationship between proline accumulation and salt tolerance in
tomato plants. In the present study, despite that the most
salt-tolerant cultivars, Brillante and Jaguar, registered
the highest foliar concentrations in proline (Table 3),
the relationship of this with biomass production indicates its low reliability for us in selection and breeding
programs against salt stress (r = 0.60 NS).
Like proline, sugars have raised controversy in recent years regarding their role in saline stress. Although
the accumulation of soluble carbohydrates in plants
has been widely reported as a response to salinity or
drought, despite a significant decrease in net CO2 assimilation rate (Murakeozy et al., 2003; Ashraf and
Harris, 2004), other studies do not support this response
(Ashraf, 1997). In different tomato cultivars studied in

Table 3
Foliar concentration of proline, sucrose and total phenols in 10
commercial cultivars of tomato plants subjected to salinity stress
(100 mM NaCl)
Cultivar
Cencara
Tesoro
Sesenta
Marimba
Caramba
Zinac
Daniela
Royesta
Brillante
Jaguar
LSD0.05
Significanceb

Proline
aba

630.92
499.70 b
345.64 c
434.04 b
176.71 e
268.99 d
474.73 b
272.98 d
730.65 a
693.33 a
70.22
***

Sucrose

Total phenols

5.35 cd
5.82 c
5.51 cd
5.57 cd
5.18 cd
4.95 d
5.66 cd
5.04 d
8.97 a
7.52 b
0.75
***

826.54 ab
786.10 bc
784.74 bc
829.57 ab
763.43 bc
719.28 b
695.73 c
892.87 a
814.40 ab
757.38 bc
89.42
**

Concentration of proline and total phenols expressed as g g1 FW.

Concentration of sucrose expressed as mg g1 FW.
a Values followed by the same letters within each column are not
different using the Duncans multiple range test at 95%.
b Levels of significance are represented by (*) at P < 0.05, (**) at
P < 0.01, (***) at P < 0.001 and NS: not significant.

our work, the highest sucrose concentration appeared

in the most salt-resistant cultivars (Table 3), whereas
the lowest were presented by the most salt-sensitive
variety (Table 3). The relationship of sucrose with the
concentrations of Na+ and Cl as well as with biomass
indicate, that this compatible compound can be used as
an indicator of salt tolerance (sucroseNa+ concentration, r = 0.77 **; sucroseCl concentration, r =
0.80 **; sucrosefoliar biomass, r = 0.83 ***).
Another group of compounds that has been related
to the response of diverse types of biotic and abiotic
stress are phenols (Ruiz and Romero, 2001). The foliar concentrations of these secondary compounds in
the present experiment varied with respect to Na+ and
Cl (Na+ concentrationtotal phenols, r = 0.54 NS;
Cl concentrationtotal phenols, r = 0.48 NS) as the
highest values were presented by the cv. Cencara and
the lowest in cv. Daniela (Table 3), thus due to inconsistent relationship between these compounds and foliar
biomass (r = 0.24 NS), the use of phenols as a selection criterion for salt tolerance be with caution.
Finally, in recent research, the decrease in lipid peroxidation and the increase in thiols such as glutathione
(GSH) have proven significance in responses to different types of stresses(Hofgen et al., 2001; Mittler,

M. Juan et al. / Environmental and Experimental Botany 54 (2005) 193201

Table 4
LOX activity, MDA content and total SH content in 10 commercial
cultivars of tomato plants subjected to salinity stress (100 mM NaCl)
Cultivar

LOX activity

MDA

Total SH content

Cencara
Tesoro
Sesenta
Marimba
Caramba
Zinac
Daniela
Royesta
Brillante
Jaguar
LSD0.05
Significanceb

0.33 ca
0.32 c
0.31 c
0.38 c
0.59 b
0.61 b
0.32 c
0.12 a
0.21 d
0.19 d
0.07
***

42.82 b
44.73 b
43.78 b
44.56 b
42.45 b
43.47 b
43.88 b
54.90 a
33.67 c
35.92 c
6.74
***

0.33 b
0.26 c
0.31 bc
0.31 bc
0.27 c
0.29 c
0.28 c
0.16 d
0.47 a
0.39 b
0.07
***

LOX activity expressed as U mg1 protein; MDA content expressed

as nmol g1 FW. Total SH content expressed as mol g1 FW.
a Values followed by the same letters within each column are not
different using the Duncans multiple range test at 95%.
b Levels of significance are represented by (*) at P < 0.05, (**) at
P < 0.01, (***) at P < 0.001 and NS: not significant.

2002). Several studies with different plant species have

concluded that lipid peroxidation in salt-sensitive lines
increased more than in the salt-tolerant lines under salt
stress. In addition, some studies indicate that high levels of antioxidants and an active ascorbate-glutathione
cycle are associated with salt tolerance (Gossett et al.,
1994; Dionisio-Sese and Tobita, 1998; Bor et al., 2003).
Specifically, in tomato, Shalata and Tal (1998) assessed
the possible involvement of the antioxidant system in
salt tolerance. In the present work, lipid peroxidation
measured by the activity of LOX and the quantity
of MDA were lowest in tolerant cultivars with lower
concentrations of Na+ and Cl (cv. Brillante and cv.
Jaguar), while greater lipid peroxidation and therefore,
more oxidative stress occurred in the most sensitive
variety (cv. Royesta; Table 4).
In our experiment, we found the harmful effect of
excessive accumulation of Na+ and Cl under saline
conditions to be accompanied by increasing lipid peroxidation (Na+ concentrationLOX activity, r = 0.91
***; Na+ concentrationMDA content, r = 0.90 ***;
Cl concentrationLOX activity, r = 0.89 ***; Cl
concentrationMDA content, r = 0.96 ***), thereby
inhibiting biomass production and thus reduced plant
adaptation to stress (LOX activityfoliar biomass, r =
0.85 ***; MDA contentfoliar biomass, r = 0.87
***).

199

The quantity of thiol groups (SH-), mainly GSH,

may be determinant in tolerance of stressed plants.
Metabolites reportedly increase under various types
of environmental stresses, their main function being
to reduce oxidative damage under such stress conditions (Hernandez et al., 1993, 1995; Gueta-Dahan et
al., 1997; Hofgen et al., 2001). Table 4 reveals that the
most tolerant tomato cultivars had the least lipid peroxidation (cv. Brillante and cv. Jaguar). They also had the
greatest capacity to synthesize these compounds, registering the highest concentrations in total thiol groups.
By contrast, the most sensitive cultivar, with the highest lipid peroxidation (cv. Royesta), presented the lowest thiol concentrations (Table 4). The relationship between biomass production and concentration of the total thiol groups (r = 0.86 ***) reflects the great reliability of this bioindicator in selection and breeding
programs for enhanced salt tolerance.
Similarly, the increase in carotenoids in tolerant
plants (cv. Brillante and cv. Jaguar) could protect these
cultivars against salinity stress, given that these pigments are responsible for quenching singlet oxygen and
thereby avoiding lipid peroxidation and consequent oxidative damage (Knox and Dogge, 1985; Sairam et al.,
2002).
In conclusion, in the present work, we propose the
use of certain nutritional and biochemical indicators
for salt tolerance in tomato, such as la reduction in
uptake and foliar accumulation of Na+ and Cl , increasing K+ uptake, and greater synthesis of sucrose,
carotenoids, and thiol groups, as well as the reduction
fundamentally of lipid peroxidation. This reflects the
importance in tomato plants of oxidative damage as
an effective physiological process in the selection of
salinity-tolerant tomato plants.

Acknowledgements
This work was supported by the project CO1-139
(Junta de Andaluca).

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