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In ternationa l
Scholars
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African Journal of Agricultural Marketing ISSN: 2375-1061 Vol. 12 (6), pp. 001-006, June, 2024. Available 
online at www.internationalscholarsjournals.org © International Scholars Journals 

 

Author(s) retain the copyright of this article. 
 
 

 

Full Length Research Paper 

 

Impact of Salinity on Seed Germination and Early 
Seedling Development in Safflower (Carthamus 

tinctorius L.) 
 

Mostafavi Khodadad 

 
Islamic Azad University- Karaj Branch, Iran. E-mail: mostafavi@kiau.ac.ir. Tel: 00989365686610. 

 
Accepted 21 January, 2024 

 
In order to study the effect of salinity stress on germination and early seedling growth of six safflower 
genotypes namely KM5, KM8, KM12, KM19, KM47 and Kose by using five concentrations of NaCl (0, - 
0.3, -0.5, -1 and -1.5 MPa) a factorial experiment was designed using Completely Randomized Design 
(CRD) with three replications in Biotechnology Laboratory, Islamic Azad University-Karaj Branch. 
Results of ANOVA showed that salt stress adversely affected the germination percentage, germination 
rate, shoot length, root length, seedling length, root/shoot length ratio, seed vigour, and germination 
index and mean germination time of all 6 genotypes of safflower, which demonstrates high diversity 
among genotypes that enabled us to screen salinity tolerant cultivar. At the highest salt level (-1.5 MPa), 
Kose produced maximum germination percentage and germination rate of all genotypes and they were 
considered as relatively tolerant. Best level of NaCl concentration in root length, shoot length, seedling 
length and seed vigour was -0.3 MPa. Seed vigour increased with increase in osmotic potential until -0.3 
MPa but decreased in -0.5 MPa. Results of cluster analysis (Ward’s minimum variance method) at the 
highest salt level (-1.5 MPa) classified all genotypes into three group. According to the obtained results, 
we found that Kose is the most resistant and KM5, KM8 and KM47 are the most sensitive genotypes. 

 
Key words: Cluster analysis, germination indices, NaCl, seed vigour. 

 
 
INTRODUCTION 

 
Among various environmental stresses, soil salinity has 
become a critical problem worldwide due to its dramatic 
effects on plant physiology and performance (Golbashy et 
al., 2010). Salinity in soil or water is one of the major 
stresses and especially in arid and semi arid regions, can 
severely limit crop production (Shannon, 1998). Breeders 
seek to develop and identify cultivars that are more 
tolerant of salinity and water stress (Janmohammadi et 
al., 2008). Germination is generally considered to be the 
developmental stage that is most salt-sensitive, 
especially for crops exposed to hostile environments 
(Ashraf and Wahid, 2000).  

Salinity impairs seed germination, reduces nodule 
formation, retards plant development and reduces crop 
yield (Greenway and Munns, 1980). Soil salinity may 
affect the germination of seeds either by creating osmotic 
potential external to the seeds preventing water uptake or 

 
 
 
 

 
through the toxic effects of Na+ and Cl- ions on 
germinating seed (Golbashy et al., 2010; Khajeh-Hosseini 
et al., 2003; Atak et al., 2006; Kaya et al., 2006).  

Salinity delays the onset, reduces the rate and 
increases the dispersion of germination events, resulting 
in reduced plant growth and final crop yield (Ashraf and 
Foolad, 2005). Absence of germination in salinity soil is 
very often due to the high concentration of salt in the soil 
where the seeds are sown. The reason is that the salt 
solution moves upward, following the evaporation at soil 
level (Bernstein, 1974). Salt disturbs both germination 
and plant growth (Fowler, 1991). The main salt-induced 
physiological disorder is diminished seed imbibitions 
because of the low solute potential within the saline 
growth medium (Debez et al., 2004). Seed may be more 
sensitive to stress than mature plants because of 
exposure the dynamic environment close to the soil 



2 

 

 
 
 

 

surface. One of the commonest experiments in 
germination of the seeds is the application of NaCl. Seed 
response to salinity can be simulated by NaCl induced 
ionic stress in the germination experiments. Ionic stress is 
caused by a toxic accumulation of NaCl in plant tissues. 
Germination rates decrease with an increase in NaCl 
concentration (Murillo-Amador et al., 2002).  

Thus, the salt-affected soils can be utilized by growing 
salt tolerant plants, whether halophytes or crops (Siddiqi 
et al., 2007). With this fact in mind, it is imperative to 
explore intra-specific (inter-cultivar) variation for salt 
tolerance of a crop by screening its available germplasm. 
For instance, a great magnitude of inter-cultivar variation 
for salt tolerance has been observed in different species 
such as wheat (Ashraf and McNeilly, 1988), lentil (Ashraf 
and Waheed, 1993), barley (Belkhodja et al., 1994), 
cotton (Ashraf and Ahmad, 1999), Brassica napus (Ulfat 
et al., 2007) and Safflower (Siddiqi et al., 2007). Safflower 
(Carthamus tinctorius L.) is one of the prospective oil-
seed crops, because it yields about 32 to 40% seed oil 
(Weiss, 1983). However, due to its considerable salt 
tolerance compared with commonly grown oil-seed crops, 
it is usually cultivated in arid and semi-arid regions where 
soil salinity is one of the major threats to agriculture 
(Kaya, 2009).  

The research has shown that in response to soil 
salinity, seedlings growth, leaves area, root biomass and 
shoot biomass have all been reduced (Redmann et al., 
1994). Although salt stress adversely affects the growth 
of safflower plants at all developmental stages (Kaya et 
al., 2003; Jamil et al., 2006; Golbashy et al., 2010), 
varietal differences in salt tolerance of safflower have 
been observed at germination (Ghorashy et al., 1972), at 
adult (Ashraf and Fatima, 1995) as well as at both 
germination and adult growth stages (Francois and 
Bernstein, 1964). However, Kaya et al. (2006) reported 
that germination percentage was not influenced by NaCl 

level of 23.5 dsm
-1

. Mohammed et al. (2002) reported 

that by NaCl levels germination percentage decreased 
and mean germination time increased proportionately.  

The present study was therefore, conducted with the 
objectives to determine the response of safflower 
genotype to salinity stress at germination and seedling 
stages under controlled conditions. Moreover, NaCl was 
used for salinity stress induction in safflower. 
 
 
MATERIALS AND METHODS 
 
In order to study the effects of salinity stress on germination and 
early seedling growth in safflower genotypes, an experiment was 
conducted in factorial form, using a completely randomized design 
with three replications. In this experiment, six safflower genotypes 
inclusive KM5, KM8, KM12, KM19, KM47 and Kose were evaluated 
in five levels of salinity treatment (distilled water as control, -0.3, - 
0.5, -1 and -1.5 MPa) by using different NaCl concentrations. This 
experiment was carried out at Biotechnology Laboratory, Islamic 
Azad University- Karaj Branch.  

The seeds were sterilized by soaking in a 5% solution of 
hypochlorite sodium for 5 min. After the treatment, the seeds 

 
 
 
 

 
were washed several times with distilled water. 25 seeds were put 
in each petridish (with 9 cm diameter) on filter paper moistened with 
respective treatment in 3 replications. The petridishes were covered 
to prevent the loss of moisture by evaporation. The petridishes were 
put into an incubator for 12 days at a temperature of 25°C and 65% 
relative humidity. Every 24 h after soaking, germination percentage 
and other traits were recorded daily. After 12 days of incubation, 
shoot length, root length, seed vigour and root to shoot ratio of 
germinated seeds was measured. Seeds were considered 
germinated when the emergent radical reached 2 mm length. 
Germination percentage, germination rate and seed vigour were 
calculated using the following formulas:  
 
Formula 1:   
 
where GP is germination percentage, SNG is the number of 
germinated seeds, and SN0 is the number of experimental seeds 
with viability (Close and Wilson, 2002; Danthu et al., 2003).  
 
Formula 2:   
 
where: GR: Germination rate; n: number of germinated seed on gth 
day and g: Number of total germinated seeds. 

 
Formula 3: Seed vigour = [seedling length (cm) × germination 
percentage] 
 
Analysis of variance was performed using standard techniques and 
differences between the means were compared through Duncan 
multiple range test (P < 0.05) using SAS release 9.1 (SAS, 2002) 
software package. All investigated traits were subjected to 
hierarchical cluster analysis using procedure ward’s minimum 
variance method as a clustering algorithm using Stat Graphics Plus 
(Ver 2.1) software. Ward’s minimum method is a hierarchical 
clustering procedure in which similarity used to join clusters is 
calculated as the sum of squares between the two clusters summed 
over all variables (Hair et al., 1998). It minimizes them within cluster 
sums of squares across all partitions. 
 

 

RESULTS 

 

Analysis of variance showed that, there were significant 
difference between genotypes, salinity stress levels and 
their interaction. The results of this study reveal that 
various concentrations of NaCl had a significant effect on 
the all measured traits (Table 1). The control showed 
clear genetically differences among the genotypes 
regards germination percentage, and such differences 
were statistically significant.  

Germination percentage of all safflower genotypes was 
adversely affected due to the application of different 
levels (0, -0.3, -0.5, -1 and -1.5 MPa) of NaCl.  

Also analysis of variance showed that, interaction 
effects was significant for all investigated characters 
except root to shoot length ratio and mean germination 
time.  

The differences between the means (Genotypes and 
salinity stress levels) were compared by Duncan multiple 
range test and are shown in Table 2. It observed that, in 
all of genotypes there was a decrease in germination 
percentage due to salinity stress increment and maximum 
germination percentage was delayed. While in 



3 

 

 
 
 

 
Table 1. Analysis of variance of measured traits of safflower genotypes under salinity stress.  
 

S.O.V df 
Germination 

Germination rate 
Root length Shoot length Seedling length 

 

percentage (mm) (mm) (cm)  

   
 

Genotype 5 2325.237** 1995.124** 13.099** 28.818** 79.357** 
 

stress 4 680.840** 1039.946** 29.224** 57.193** 167.838** 
 

Genotype× stress 20 167.547* 85.336** 2.871** 3.143** 10.888** 
 

error 60 83.944 23.827 1.053 0.834 3.003 
 

 

S.O.V df Seed vigour 
Root/Shoot Germination 

Mean germination time (day)  

length (mm) Index  

    
 

Genotype 5 701732.309** 1.122** 2594.791** 1.597** 
 

stress 4 1268344.148** 0.020ns 1043.178** 1.353** 
 

Genotype× stress 20 93812.719** 0.140ns 110.124** 0.046ns 
 

error 60 22757.840 0.149 44.722 0.033 
 

 
*, **, ns: significant at 5%, 1% level and not significant, respectively. 
 
 

 
Table 2. Mean comparison of main effects using Duncan multiple range test (at 5% probability level).  
 
 

Genotype 
Germination 

Germination rate 
Root Length Shoot length Seedling length 

 

 
percentage (mm) (mm) (cm)  

   
 

 KM12 76.443c 49.973c 3.021ab 4.080a 7.102a 
 

 KM19 69.334d 50.783bc 2.393ab 3.300b 5.693b 
 

 KM47 64.445d 43.133d 0.624c 0.316c 0.940c 
 

 KM5 86.667b 52.550bc 3.181a 4.073a 7.254a 
 

 KM8 69.778cd 54.139b 2.218b 2.869b 5.088b 
 

 Kose 97.333a 76.807a 2.830ab 2.888b 5.718b 
 

 Salinity stress (MPa)      
 

 0 81.853a 60.774a 3.251a 4.402a 7.653a 
 

 -0.3 78.518a 60.452ab 3.757a 4.713a 8.471a 
 

 -0.5 81.112a 57.261b 2.565b 3.096b 5.662b 
 

 -1 78.518a 51.703c 1.797c 2.011c 3.809c 
 

 -1.5 66.666b 42.631d 0.518d 0.382d 0.901d 
 

 
Genotype Seed vigour 

Root/shoot Germination 
Mean germination time (day)  

 length (mm) index  

     
 

 KM12 579.480a 0.734ab 57.267c  2.032b 
 

 KM19 418.130b 0.776ab 51.867d  2.035b 
 

 KM47 62.170c 0.200c 45.600e  2.360a 
 

 KM5 662.620a 0.714b 66.067b  1.937bc 
 

 KM8 368.250b 0.757ab 53.600cd  1.888c 
 

 Kose 568.740a 1.038a 82.533a  1.360d 
 

 Salinity stress (MPa)      
 

 0 651.900a 0.716a 64.889a  1.752c 
 

 -0.3 713.820a 0.677a 62.833ab  1.727c 
 

 -0.5 477.550b 0.717a 63.944ab  1.823c 
 

 -1 313.010c 0.744a 59.389b  1.981b 
 

 -1.5 59.870d 0.661a 46.389c  2.392a 
 

 
Values in a column bearing different superscript are significantly different at 0.05 levels. 



4 

 

 
 
 

 
Table 3. Supplementary analysis of interaction effects.  

 
Salinity level Germination Germination 

Root length (mm) Shoot length (mm) 
Seedling length 

 

(MPa) percentage rate (cm)  

  
 

0 624.578** 924.364** 4.087** 8.664** 20.842** 
 

-0.3 981.697** 482.971** 11.001** 19.700** 59.314** 
 

-0.5 365.936** 580.374** 6.213** 8.559** 28.480** 
 

-1 626.218** 254.259** 3.081* 4.025** 13.234** 
 

-1.5 396.999** 94.502** 0.199ns 0.444ns 1.037ns 
 

 
Salinity level 

Seed vigour Root/shoot length (mm) 
Germination 

Mean germination time (cm)  

(MPa) index  

   
 

0 245031** 0.140ns 739.289** 0.493** 
 

-0.3 498372** 0.343ns 910.633** 0.384** 
 

-0.5 227274** 0.153ns 462.989** 0.391** 
 

-1 102251** 0.537** 541.922** 0.246** 
 

-1.5 4054.980ns 0.508** 380.456** 0.269** 
 

 
*, **, ns: significant at 5%, 1% level and not significant, respectively. 

 
 

 

this experiment different genotypes had different 
response to the salinity stress. Among the safflower 
genotypes, Kose had the highest germination percentage 
and germination rate of 97.33% and 76.80 respectively.  

However, maximum reduction in germination 
percentage was observed at the highest level that is, -1.5 
MPa of NaCl. At the highest salt level (-1.5 MPa), Kose 
produced maximum germination percentage and 
germination rate of all genotypes and they were 
considered as relatively tolerant.  

Results of means comparison, using Duncan multiple 
range test, showed that germination percentage and 
germination rate were decreased by an increase in 
osmotic potential, while the maximum germination rate 
and percentage were obtained at 0 Mpa level (control 
treatment). Some studies referred that stress can 
contribute to improve germination rate and seedling 
emergence in different plant species by increasing the 
expression of aquaporins (Gao et al., 1999), 
enhancement of ATPase activity, RNA and acid 
phosphathase synthesis (Fu et al., 1988), also by 
increase of amylases, proteases or lipases activity 
(Ashraf and Foolad, 2005).  

Imposition of varying levels of NaCl significantly 
reduced all measured traits of all 6 investigated 
genotypes.  

Root length is one of the most important characters for 
salinity stress because roots are in contact with soil and 
absorb water from soil. For this reason, root length 
provides an important clue to the response of plants to 
salinity stress. A marked reduction in root length, shoot 
length and seedling length of all genotypes of safflower 
was observed due to salt stress.  

Among the genotypes, the longest root length was 
commonly determined in genotypes KM5, KM12, Kose 

 
 
 

 

and KM19 while KM47 gave the shortest root length. 
Generally, increasing salinity levels decreased root 
length, and KM5 genotype exhibited the greater 
performance in respect of root length. Result of this study 
showed that, shoot length diminished with increasing 
salinity levels in all genotypes (Table 2). The highest and 
the lowest seedling length were observed in KM5 and 
KM47 genotypes, respectively (Table 2).  

The most effective levels in reducing these attributes 
were -1 and -1.5 MPa of NaCl (Table 3). Best level of 
NaCl concentration in root length, shoot length, seedling 
length and seed vigour was -0.3 MPa. Seed vigour 
increased with increase in osmotic potential until -0.3 
MPa but decreased in -0.5 MPa (Figure1).  
A significant inter-genotype variation was observed 

under salt stress. Of all genotypes, KM5, KM12 and Kose 
produced highest seed vigour at all salt regimes, but 
lowest seed vigour was recorded in KM47 while the 
remaining genotypes were moderate in this attribute.  

Variation in the set of genotypes about root to shoot 
length ratio was not possible to discern at lower external 
salt levels, however, genotypes differed significantly at 
the two higher salt levels, that is, -1 and -1.5 MPa of NaCl 
(Table 3).  

In addition, it was clearly determined that there were no 
statistical differences between measured genotypes at 
high salinity levels (-1.5 MPa) for root length, shoot 
length, seedling length and seed vigour traits (Table 3).  

Cluster analysis was done using the data for all 
measured traits at the highest salt level (-1.5 MPa), 
because this salt level was found very effective in 
discriminating the genotypes. Results of cluster analysis 
(Ward’s minimum variance method) showed that 
genotypes Kose was found to be tolerant, while KM5, 
KM8 and KM47 sensitive to salt (Figure 2). 



5 

 

     
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

Figure 1. Seed vigour of safflower genotypes under different salinity stress.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
Figure 2. Cluster analysis of safflower genotypes under -1.5 level of salinity stress 
using Ward’s minimum variance method. 

 

 

Ajmal Khan and Weber (2006) found that, resistance to 
stress at germination stage and primary growth of 
seedling is independent from next growth stages and 
evaluation of stress tolerance need more experiment at 
next growth stages. 
 

 

DISCUSSION 

 
Screening of available germplasm of a crop is a feasible 
means of identifying salt tolerant genotypes or genotypes 

 
 

 

which could maintain a comparatively reasonable yield on 
salt affected soils (Ashraf and McNeilly, 1987). For the 
latter crops, it is advisable to assess degree of salt 
tolerance at each growth stage. In the present study, 
genotype Kose was found to be tolerant, while KM5, KM8 
and KM47 sensitive to salt. Ranking of the genotypes 
was done using the data for all measured traits at the 
highest salt level (-1.5 MPa), because this salt level was 
found very effective in discriminating the genotypes. 
These results can be related to some earlier studies in 
which genotypes identified as salt tolerant at the earlier 



6 

 

 
 
 

 

growth stages showed tolerance when tested at the later 
growth stages.  

Although a considerable magnitude of variation for salt 
tolerance was observed in a set of 6 available genotypes 
of safflower while screening them at germination stages, 
but a further study needs to be carried out to assess 
whether the genotypes marked as salt tolerant at the 
initial growth stages, maintain their degree of salt 
tolerance when tested as adult. 
 

 

Conclusion 

 

In the present study, salt stress adversely affected the 
germination percentage, germination rate, shoot length, 
root length, seedling length, and root to shoot length ratio, 
seed vigour, and germination index and mean 
germination time of all 6 genotypes of safflower and a 
significant variation in salt tolerance was observed among 
all the safflower.  

Many researchers have reported similar results (Demir 
and Aril, 2003; Mauromicale and Licandro, 2002). 
Obviously, acceptable growth of plants in arid and 
semiarid lands which are under exposure of salinity 
stress is related to ability of seeds for best germination 
under unfavourable conditions, so necessity of evaluation 
of salinity resistance genotypes is important at primary 
growth stage. To find the best tolerant genotype to such 
conditions, taking all traits into account in this study, we 
found that Kose is the most resistant and KM5, KM8 and 
KM47 are the most sensitive genotypes. 
 

 
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