EurAsian Journal of BioSciences

Eurasia J Biosci 6, 47-59 (2012)

DOI:10.5053/ejobios.2012.6.0.6

Selection of a salt tolerant Tunisian cultivar of chili

pepper (Capsicum frutescens)
Kaouther Zhani1*, Mohamed Aymen Elouer1, Hassan Aloui2, Cherif Hannachi1
1Department of Horticulture and Landscape, Sousse University, High Institute of Agronomy,

4042 Chott Mariem, Tunisia

2Carthage University, Faculty of Science, Jarzouna 7021, Bizerte, Tunisia

*Corresponding author: kaoutherzheni@yahoo.fr

Abstract
Background: Salinity affects germination and seedling growth and yield of several crop species,
such as pepper. That is why this study was carried to evaluate the effects of NaCl on seed
germination, seedling growth and ionic balance of three Tunisian chili pepper (Capsicum frutescens)
cv: Tebourba, Korba and Awlad Haffouz.
Materials and Methods: The percentage of germination, the growth and the mineral contents
were measured in the three Tunisian chili pepper cv watered with water containing 0, 2, 4, 6 or 8 g
L-1 NaCl.
Results: Results showed that different salinity stress levels had significant effect on germination
percentage and germination time. In pot experiment, increasing NaCl concentration, for all cv,
induced a significant decrease on plant height, root length, leaves number, leaf area and chlorophyll
amount. The fresh and dry weights are also affected. In addition, salinity increased Na+ and Cl levels
but decreased K+ level in roots and shoots.
Conclusions: Awlad Haffouz cv had the highest K+/Na+ ratio compared to cv Korba and Tebourba
and it has showed the best response under salt stress during germination and growth stage which
lets it to be the most tolerant cv.
Keywords: Capsicum frutescens, germination, mineral nutrition, salinity, shoot.
Zhani K, Elouer MA, Aloui H, Hannachi C (2012) Selection of a salt tolerant Tunisian cultivar of chili
pepper (Capsicum frutescens). Eurasia J Biosci 6: 47-59.
DOI:10.5053/ejobios.2012.6.0.6

INTRODUCTION
Salinity is one of the most important abiotic
stresses limiting crop production in arid and
semiarid regions, where soil salt content is high and
precipitation is low (Neumann 1995). Transpiration
and evaporation from the soil surface, salt load in
irrigation water, over use of fertilizers and lack of
proper drainage can be the main factors that
contribute to this problem. Around 930 million ha of
land world-wide, 20% of total agricultural land, are
affected by salinity (Munns 2002). Salinity limits
crops production, especially the sensitive ones
(Zadeh and Naeini 2007) and reduces the yield of
major crops by more than 50% (Bray et al. 2000). It
affects
morphological,
physiological
and
biochemical processes, including seed germination,
plant growth and water and nutrient uptake
(Willenborg et al. 2004). These effects can be due to
low osmotic potential of soil solution, specific ion
effects, nutritional imbalance or a combined effect
of all these factors (Marschner 1995). NaCl is the
predominant salt causing salinization and it is
expected that plants have involved mechanisms to

EurAsian Journal of BioSciences

regulate its accumulation (Munns and Tester 2008).

Pepper is widely cultivated for its fruits which
have a recognized nutritional value. In fact, they are
an excellent source of various antioxidant
compounds like flavonoids, carotenoids and vitamin
C (Chuah et al. 2008). This later protects human body
against oxidative damage and prevents various
diseases such as cancer and cardiovascular diseases
(Oboth and Rocha 2007). In Tunisia, Pepper is the
major cultivated plant and its fruits are mainly
consumed either fresh or dry. It is cultivated on open
air and under greenhouse. However, pepper is
exposed to many biotic (virus, fungi) and abiotic
stress, especially salinity, which has a negative effect
on pepper growth and yield (Ibn Maaouia-Houimli et
al. 2011).
The objective of this research was to study the
effect of salt stress on some characteristics of three
Tunisian chili pepper cv: Tebourba, Korba and Awlad
Haffouz by measuring seed germination, seedling
Received: April 2012
Accepted: April 2012
Printed: June 2012

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EurAsian Journal of BioSciences 6: 47-59 (2012)

growth and ionic balance at various concentrations

of NaCl (0, 2, 4, 6 and 8 g L-1) in order to select salt
tolerant cultivars.
MATERIALS AND METHODS
Seed germination assay
Seeds of three chili pepper cv (Tebourba, Korba
and Awlad Haffouz) were collected from plants
cultivated one year ago in the experimental station
of Agronomic High Institute of Chott Mariem. The
seeds were sterilized for 20 min in sodium hypochloride (5%) and then they were rinsed 3 times with
distilled water for 2 min. After sterilization, under
laminar flow, 10 seeds of each cv were transferred
into sterile Petri dishes (100x100 mm dimensions)
between two layers Watman filter paper and then
wetted with 10 mL distilled water (control) or saline
solution containing 2, 4, 6 and 8 g L-1 NaCl and left to
germinate at 25C. Germinated seeds were recorded
during 20 days.
Germination (%)= n/N x 100
n: number of germinated seeds on the nth day
N: total number of seeds
Treatments were assessed in factorial
experimental based on a completely randomized
design at 3 replications. Each replication includes
one Petri dish (ten seeds per Petri dish).
Growth assay
Seeds were sterilized for 20 min in sodium hypochloride solution (5%) and then rinsed 3 times with
distilled water. Five seeds from each cv were sowed
in plastic pot (12 cm diameter and 22 cm height)
containing gravel and fertilized peat (1/4: 3/4) at 1
cm depth. Pots were put in greenhouse under
25/18C day/night temperature and natural light.
After emergence, one seedling per pot was
conserved. For 60 days, plants were watered with
water (control) or a saline solution containing 2, 4, 6
and 8 g L-1 NaCl. The plant height (cm), root length
(cm), leaves number per plant, leaf area (cm), fresh
weight for both shoot and root were measured. Dry
weights were measured after drying into oven at
80C for 48 h.
Leaf area was measured by planimeter (Area
Meter 3100). Chl (a and b) were determined

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Zhani et al.

according to Arnon (1949) method. Samples of fresh

leaves (0.1 g) were ground with sand and 10 mL of
acetone in a mortar. The absorbance of the extracts
was measured by spectrophotometry at 645 and 663
nm.
The chl amounts were calculated according to the
following equations:
Chl a (g g-1 F.W.): 12.7 (OD 663) - 2.63 (OD 645)
Chl b (g g-1 F.W.): 22.9 (OD 645) - 4.86 (OD 663)
Chl (a+b) (g g-1 F.W.): 8.02 (OD 663) - 20.2 (OD
645)
K+, Na+ and Cl- root and shoot content were
analyzed by flame spectrophotometer. Pots were
disturbed in completely randomized design with 3
replications.
Data analysis
All data were analyzed by SPSS software 13.00
and Duncans multiple range tests were used to
determine significance between variables (P<0.05).
RESULTS AND DISCUSSION
Seed germination percentage and germination
time
Effect of NaCl concentration on the percentage
of germination in the three cv during 20 days is
shown in Figs. 1-3. In control treatment, seeds
germinated after two (Korba cv) or three days
(Tebourba and Awlad Haffouz cv) and total
germination ended after 13 (Korba and Awlad
Haffouz cv) or 14 days (Tebourba cv). Salt stress
modified germination process by increasing
germination time and decreasing germination
percentage. Under the highest concentration of
NaCl, seeds germinated after six (Korba cv) to ten
days (Tebourba cv). The maximum germination
percentage was observed in Korba cv (50%) and the
lowest one in Tebourba cv (10%). Zhani (2009) had
obtained similar results in other Tunisian pepper cv
where NaCl decreased germination percentage to
70 and 20% respectively in Starter and Chergui cv at
6 g L-1 NaCl. Keshavarzi (2011) and Keshavarzi et al.
(2011) have found the same result respectively on
some savory and spinach cv.
NaCl affected seed germination by creating
external osmotic potential which causes difficulties

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EurAsian Journal of BioSciences 6: 47-59 (2012)

Fig. 1. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on the percentage of germination of Tebourba ch ili pepper cultivar.

Fig. 2. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on the percentage of germination of Korba ch ili pepper cultivar.

in absorption of the necessary water quantities for

the germination process (Abdelly 1992). Guerrier
(1984) reported that salt stress causes also specific
toxicity by higher accumulation of Na+ and Cl- ions in
the embryo in addition to a mineral imbalance. In
extreme case, death of embryo can take place due to
an inhibition of metabolic process (Bliss et al. 1986).
Plant height
The mean of plant height varied between 36.5
(control) to 17.9 cm (8 g L-1 NaCl) (Table 1). The
longest plant height was observed in the control of
Tebourba cv (38.2 cm). When NaCl concentration
increased, plant height of Awlad Haffouz, Tebourba

and Korba cv decreased significantly at NaCl 8 g L-1

respectively to 37, 57 and 58%. The same result was
obtained in canola (Bybordi 2010) where the height
of plants declines from 56.25 cm in control to 28.19
cm at NaCl 6 g L-1.
Root length
Mean of root length was between 15.7 (control)
and 7.4 cm (8 g L-1 NaCl). As expected, control
condition and the highest NaCl level (8 g L-1) induced
the longest and the shortest root length
respectively (Table 1). Generally, root length
decreased as salt stress increased. In the control, it
varied from 14.1 (Korba cv) to 18.8 cm (Awlad
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EurAsian Journal of BioSciences 6: 47-59 (2012)

Fig. 3. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on the percentage of germination of Awlad Haffouz ch ili pepper cultivar.
Table 1. Plant height (cm) and root length (cm) of three ch ili pepper cultivars watered during 60 days with water
containing NaCl 0, 2, 4, 6 or 8 g L-1.

Means followed by the same letter(s) are not significantly different at P= 0.05 according to Duncan test.

Haffouz cv). At the highest stress level, Korba and

Awlad Haffouz cv showed a respective decrease of
58 and 50% where root length was 7.02 and 9.3 cm
respectively. Kerkeni (2002) obtained a similar result
in potato.
Leaf characteristics
Mean number of leaves per plant (Fig. 4) showed
a decrease with the increase of salt stress in all chili
pepper cv. At highest NaCl concentration (8 g L-1),
pepper plant didnt produce more than 9 leaves
(Korba cv) which correspond to 81% decrease
compared to control (47 leaves per plant). The result
agrees with the report of Mensah et al. (2006) in
groundnut where it was observed that salinity at 17
mS/cm enhanced the production of leaves in RMP91
cv from 42.7 (control) to 19.3 leaves.
50

According to Fig. 5, when NaCl increased, leaf

area decreased to 78% for Awlad Haffouz cv with
NaCl 8 g L-1. Studies done on five cultivars of canola
(Bybordi 2010) gave similar results, leaf area of
canola decreased from 256.25 cm in control to
107.31cm with NaCl 6 g L-1.
Chl a, b and a+b amounts in leaves were the
highest in control (Figs. 6-8). Tebourba cv leaves
were the richest (2.875 g g-1 F.W.) whereas Awlad
Haffouz cv leaves were the poorest (2.282 g g-1
F.W.). NaCl decreased chl (a+b) synthesis in the three
chili pepper cv and this decrease was the most
important at the highest NaCl concentration (8 g L-1);
chl a+b decrease was 55% in Tebourba cv and 66% in
Awlad Haffouz cv. The same trend was obtained for
chl a and chl b amounts but the response of the cv

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Zhani et al.

Fig. 4. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on the number of leaves per 60 days old plant of three ch ili pepper cultivars.

Fig. 5. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on leaf area (cm2) of three 60 days old ch ili pepper cultivars.

was different. At NaCl 8 g L-1, leaves of Tebourba cv

had the highest amounts of both chl a (0.84 g g-1
F.W.) and chl b (0.4 g g-1 F.W.) while the lowest

amounts were observed in Korba cv (0.38 g g-1 F.W.)

and Awlad Haffouz cv (0.21 g g-1 F.W.) respectively
for chl a and chl b. Biricolti and Pucci (1995) observed
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Zhani et al.

Fig. 6. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on the chlorophyll a content (g/g FW) in three 60 days old ch ili pepper cultivars.

Fig. 7. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on the chlorophyll b content (g/g FW) in three 60 days old ch ili pepper cultivars.

such result in peach where chl a synthesis was

reduced in Readheaven cultivar by salt treatment.
Agastian et al. (2000) reported that at higher salinity
(12 mS/cm) chl a was totally eliminated in mesophyll
52

of Mulberry because of total destruction of

chloroplast structure (Blumenthal-Goldschidt and
Poljakoff-Mayber 1968).

EurAsian Journal of BioSciences 6: 47-59 (2012)

Zhani et al.

Fig. 8. Effect of NaCl (0, 2, 4, 6 or 8 g L-1) on the chlorophyll a+b content (g/g FW) in three 60 days old ch ili pepper cultivars.

Fresh and dry weights

Fresh and dry weights of aerial parts and roots of
three chili pepper cv grown in 0 to 8 g L-1. NaCl are
presented in Table 2. The shoot and root fresh and
dry biomass of the three studied cv were
significantly reduced with increasing NaCl
concentration. At the highest salinity, cv Awlad
Haffouz had the highest biomass and Tebourba cv
had the lowest ones. Thus, at the highest salt
concentration, the dry weight of Tebourba cv
decreased till 88 and 92% for root and shoot
respectively.
Al Thabet et al. (2004) working on canola, Ben
Said (2004) on melon, Ibriz et al. (2005) on luzerne
and Singh et al. (2007) on groundnut indicated that
under salinity stress plant growth was inhibited
because salinity exerted low water potential, ion
toxicity and ion imbalance (Greenway and Munns
1980). In the three chili pepper cv, shoots were more
affected by NaCl than roots. Those results are similar
to those reported by Hajlaoui (2003) in chick pea,
Akinci et al. (2004) in eggplant and Saboora et al.
(2006) in wheat plants. However, Bybordi et al.
(2010) have showed that root length was the most
affected in the five studied canola cultivars.
El-Bassiouny and Bekheta (2005) have shown that
accumulation of ions in wheat plants grown in the
presence of salt (14 dS/cm) environment causes
osmotic and pseudo-drought stresses leading to
decrease of water absorption. The decrease of

tissue water content resulted in reduction of cellular

growth and development. Therefore, restriction of
water absorption was one of the most important
causes of stem and root growth decrease. Farhoudi
and Tafti (2011) reported that root cells have a much
less turgor threshold pressure than that of stem
cells thus root growth was more than stem growth
under salt and drought stresses. Therefore, root was
significantly less affected by salt stress in
comparison to stem (Sadeghi 2009).
Mineral analysis
Results in Table 3 show that in aerial parts and
roots low concentrations of Na+ and Cl- were
observed in control plants. Values for roots were
lower than those for shoots. Increasing NaCl
concentration amplified Na+ and Cl- contents in
shoots and roots in the three cv and decreased at
the same time K+ content. The present result was in
agreement with the work of Mezni et al. (2002) in
luzene, Kaya et al. (2002) in strawberry, Sahloul
(2002) in tomato, Ben Dkhil and Denden (2010) in
okra and Akbarimoghaddam et al. (2011) in wheat,
those authors observed that high saline
concentration increased Na+ and Cl- contents and
decreased K+ content in the affected crops. Bybordi
et al. (2010) showed that potassium content
decreased due to salinity in sensitive canola cv. It
seems that the decrease in potassium content is due
to an antagonistic effect between sodium and
potassium. Greenway and Munns (1980) had

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Zhani et al.

Table 2. Plant biomass of three chili pepper cultivars watered during 60 days with water containing NaCl 0, 2, 4, 6 or 8 g L-1.

Means followed by the same letter(s) are not significantly different at P= 0.05 according to Duncan test.

Table 3. Na+, Cl- and K+ concentrations (meq/g DW) in roots and shoots of three ch ili pepper cultivars watered during 60
days with water containing NaCl 0, 2, 4, 6 or 8 g L-1.

Means followed by the same letter(s) are not significantly different at P= 0.05 according to Duncan test.

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highlighted the antagonistic effect between these

two elements.
Na+ content in shoots of Awlad Haffouz cv was
significantly lower than Na+ content in the other chili
pepper cv. According to these results, it was
concluded that Awlad Haffouz cultivar was the most
salt stress tolerant due to its less Na+ absorption and

Zhani et al.

more K+ accumulation in roots compared with the

two other studied cv. Ashraf and Harris (2004)
reported that Na+ and Cl- accumulation in tolerant cv
was lower than in sensitive cv and K+ concentration
was higher in tolerant cv. Additionally, according to
results showed in Figs. 9 and 10, K+/Na+ ratio was the
highest in this cv, especially at the highest salt stress

Fi g. 9. Effec t of NaCl (0, 2, 4, 6 or 8 g L-1) on shoot K+/Na+ r at i o in t hree 60 days old chil i pepper cult ivars.

Fi g. 10. Effec t of NaCl (0, 2, 4, 6 or 8 g L-1) on r oot K+/Na+ r at i o in t hree 60 days old chil i pepper cult ivars.
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EurAsian Journal of BioSciences 6: 47-59 (2012)

concentration, which was 0.08 in shoots and 0.07 in

roots. Thus, this result explains the advantage of this
cv during its germination and its vegetative growth
in the presence of NaCl. Morant-Manceau et al.
(2004) and Farhoudi and Tafti (2011) found also that
K+/Na+ ratio was higher in salt tolerant triticale and
soybean cv respectively. The results for this tolerant
cv can be explained in the light of early findings of
many scientists that salt tolerant mesophytes
generally excluded either Na+ and Cl- from their
shoots (Lauchli et al.1994, Saqib et al. 2005) because
Na+ was the primary cause of ion specific damage,
resulting in a range of disorders in enzyme activation
and protein synthesis (Tester and Davenport 2003).
Therefore, exclusion of Na+ at root level and
maintenance of a high K+ content in the shoots were
vital for the plants to grow under saline conditions
(Munns et al. 2000). This cv also maintained
considerably high K+/Na+ ratio in both shoots and
roots. This trait has a potential value as selection
criterion for salt tolerance (Greenway and Munns
1980).

CONCLUSION
Results of this study demonstrate that NaCl
affects some of the physiological process in pepper.
The increase of salinity level decreased all studied
parameters except Na+ and Cl- concentrations in
aerial parts and in roots. Awled Haffouz cv had a
higher tolerance to salinity compared with Korba
and Tebourba cv. It is clear that the main
mechanisms for the salt tolerance of pepper were
exclusion of Na+ and Cl- from shoots, high uptake of
K+ and maintening a high K+/Na+ ratio. The
measurement of Ca+ content, organic solutes
synthesis such as proline, soluble sugars, soluble
proteins and free amino-acids would allow to better
explain the salt tolerance in pepper. Also, efficiency
of ion transporters, cellular compartmentation of
ions, oxidative stress and synthesis of osmoticums in
relation with salinity stress are worth studying.

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Zhani et al.

Tuza Dayankl Tunus Krmz Biberinin (Capsicum frutescens) Seilmesi

zet
Giri: Tuzluluk; biber gibi baz tarm bitkilerinde imlenmeyi, fide bymesini ve verimi etkilemektedir. Bu yzden bu
alma, Tunus biber (Capsicum frutescens) eidinde, Tebourba, Korba ve Awlad Haffouz, NaCln tohum
imlenmesi, fide bymesi ve iyon dengesi zerindeki etkilerini aratrmak iin gerekletirildi.
Materyal ve Metot: 0, 2, 4, 6 veya 8 g L-1 NaCl ieren suyla sulanan Tunus krmz biber eidinde imlenme yzdesi,
byme ve mineral ierikleri lld.
Bulgular: Bulgular, deiik tuz stresi seviyelerinin, imlenme yzdesi ve imlenme zaman zerinde nemli etkisinin
olduunu gstermitir. Saks deneyinde, artan NaCL konsantrasyonu, btn eitlerde bitki boyu, kk uzunluu,
yaprak says, yaprak alan ve klorofil miktarnda nemli azalmaya sebep oldu. Ya ve kuru arlklar da etkilendi. Buna
ek olarak tuzluluk, kk ve srgndeki Na+ ve Cl seviyelerini artrd, fakat K+ seviyelerini azaltt.
Sonu: Awlad Haffouz eidi Korba ve Tebourba eitlerine kyasla, en yksek K+/Na+ oranna sahipti ve imlenme
esnasndaki en iyi tuz stres tepkisini verdi. Bu durum, bu eidin en toleransl eit olduunu gstermektedir.
Anahtar Kelimeler: Capsicum frutescens, imlenme, filiz, mineral beslenme, tuzluluk.
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