ISSN: 1579-4377

SALINITY (NACL) TOLERANCE OF BRASSICA SPECIES AT GERMINATION

AND EARLY SEEDLING GROWTH

Muhammad Jamil1, *, Cheong Chun Lee1, Shafiq Ur Rehman2, Deok Bae Lee3, Muhammad Ashraf4,
Eui Shik Rha1
1
College of Agriculture and Life Sciences, Sunchon National University, Suncheon 540-742, Korea
2
College of Agriculture, Chonbuk National University, Jeonju, 561-756, Korea
3
Honam Agricultural Research Institute, NICS, Iksan 570-080, Korea
4
Department of Botany, Agriculture University, Faisalabad, Pakistan

KEYWORDS
Brassica species, germination, seedling growth, salinity (NaCl) tolerance

ABSTRACT
Salinity tolerance during germination and early seedling growth was evaluated for Brassica species such
as cabbage (Brassica oleracea capitata), cauliflower (Brassica oleracea botrytis) and canola (Brassica
napus) in four treatments of salinity including 0.0 (control), 4.7, 9.4 and 14.1 dS m-1 NaCl. The results
showed that different treatments of salinity had considerable effect on the germination, germination rate,
shoot and root length, shoot and root fresh weight, leaf area and number of leaves of canola, cabbage and
cauliflower. Germination percentage in all three species showed considerable decrease with increasing
salinity up to 14.1 dS m-1 NaCl. This reduction was more in cauliflower as compared to cabbage and
canola. The required time for germination also increased with increasing levels of salinity. The seedling
growth of three species was significantly inhibited by all salinity levels. Particularly at 14.1 dS m-1 NaCl,
root and shoot length of all species were considerably reduced. Shoots growth of all three plant species
were more affected as compared to root growth at all salinity levels. Fresh shoot and root weight, leaf
area and number of leaves were also severely affected at all salinity treatments.

INTRODUCTION
Soil salinity is one of the most important factors that limit crop production in arid and semi arid regions
[1]. The plant growth is ultimately reduced by salinity stress but plant species differ in their salinity
tolerance [2]. Salinity stress is an important characteristic when selecting a variety for salinity tolerance
[3]. Plants adapt to stress by different mechanisms, including changes in morphological and
developmental stresses as well as physiological and biochemical processes [4].

High levels of soil salinity can significantly inhibit seed germination and seedling growth, due to the
combined effects of high osmotic potential and specific ion toxicity [5]. Seed germination, seedling
emergence, and early survival are particularly sensitive to substrate salinity [6 and 7]. Salinity stress can
affect seed germination through osmotic effects [8]. Successful seedling establishment depends on the

*
Corresponding Author E-mail: jamilkhattak@yahoo.com
Present address: College of Agriculture and Life Sciences, Sunchon National University, Plant Genetics Lab. 315 Maegok,
Suncheon Jeonnam 540-742, Korea

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frequency and the amount of precipitation as well as on the ability of the seed species to germinate and
grow while soil moisture and osmotic potentials decrease [9]. Plants have different responses to salinity
depending on their stage of development [10].

Salt tolerance is important in vegetables because of the cash value of crops [11]. Importance of the
Brassica vegetables such as Chinese cabbage, broccoli, cabbage, cauliflower and kale in terms of their
nutrient quality for human diet and processed food has, recently, been increased [12]. Significant
variation was reported in seed germination between canola cultivars grown under salinity condition is
widely reported by Zheng et al. [13] and Puppala et al. [14]. The two species of canola B. napus and B.
campestris are classified as tolerant to salinity reported by Maas and Hoffman [15]. Maas [16] also
reported that even though both the species exhibit high salinity thresholds, the rate of yield decline above
the thresholds was much greater than most other crops in the tolerant category.

Salinity is an important constraint to crop production in the world. Because of salinity problems,
researchers are trying to get the salt resistant vegetables on which human’s food depend. The present
study was therefore initiated to investigate the extent of salinity on the germination and early seedling
growth of Brassica species.

MATERIALS AND METHODS

The seeds of Brassica species, cabbage (Brassica oleracea capitata L.), cauliflower (Brassica oleracea
botrytis L.) and canola (Brassica napus) differing in salt sensitivity were used in this study. Seeds of
these plant species were obtained from China.

Experiment was conducted to examine the effect of different NaCl concentrations on germination,
germination rate (1/t50, where t50 is the time to 50% of germination), root and shoot length, leaf area (leaf
area of individual plant was taken in cm2 by measuring the length and the maximum width of the main
leaf and the product (L×W) was multiplied by 0.836 as described by Dhiman and Kulra [17]) and number
of leaves per plant. Plants were planted in plastic Pots (16 cm diameter, 22 cm height) in sand culture.
The solution consisted of 0.0 (control), 4.7, 9.4 and 14.1 dS m-1 NaCl along with Hoagland’s nutrient
Solution [18]. For each plant species 10 seeds for each of the four NaCl treatments were used. Seeds were
hand sorted to eliminate broken and abnormal ones. Seed were allowed to germinate in green house
condition. Germinated seed was investigated after every 12 hours. Seeds started to germinate after 36
hours (seeds were considered to be germinated with the emergence of the radical). The germinated seeds
were observed at daily intervals. The lengths of roots and shoots of the germinated seeds which were
more than 2 mm in length were measured and recorded after 15 days of sowing. In all treatments a
continuous increase in the number of germinating seeds as well as in the lengths of roots and shoots was
observed during the frequent days of germination.

The experiment was designed by using a randomized complete block design with three replications.
Linear trend lines were fitted to transformed final germination into arcsin form [19]. Analysis of variance
was performed by using the Microsoft Excel version 5.0 [20]. Means values for different plant
characteristics were compared through LSD test [21].

RESULTS

Analysis of variance revealed that significant differences among the Brassica species for germination
percentage. Germination percentage was strongly affected by all salinity treatments. Increased salt
concentration caused a decrease in final germination percentage. There was considerable reduction in
germination parentage in canola, cabbage and cauliflower at all salinity levels particularly at 14.1 dS m-1
NaCl (Table 1). The reduction being strongest particularly at the highest level of salt concentration
compared to control. Germination response to 4.7 dS m-1 NaCl was not significantly different in case of
cabbage and canola then control. Canola showed good germination response at all salinity levels among

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all plant species. The reduction was more in cauliflower particularly at high salinity concentration as
compared to cabbage and canola. The results presented in Table 1 also show significant reduction in
germination in cauliflower at lowest concentration and at highest concentration where germination was
projected nearly to zero.

The final germination rate of seeds of these plant species under various conditions of salinity was
expressed as a 1/t50 of the germination of seeds of the same population as in control. The germination
response of the three plant species under observation showed considerable differences in the timing of
initiation and completion of germination. Germination started within 36 hours and was complete on the
6th day. Seed germination delayed as the level of salinity increased. Table 1 indicated that canola and
cabbage completed their germination nearly in same time but cauliflower took comparatively more time
to complete germination especially at high salt treatments.

Table 1. Effect of Different NaCl Treatments on the Germination (A) and Germination Rate (B) of Canola, Cabbage
and Cauliflower.

(A) Germination (Arcsin transformed) (B) Germination rate (1/t50)

NaCl (dS m-1) Cabbage Cauliflower Canola Cabbage Cauliflower Canola
0.00 90.00 90.00 90.00 0.67 0.61 0.67
4.70 90.00 68.86 90.00 0.61 0.43 0.56
9.40 68.86 35.22 68.85 0.47 0.32 0.47
14.10 21.15 18.44 50.85 0.38 0.26 0.38
LSD(0.05) 3.91 3.44 4.44 0.07 0.07 0.07

The studies were carried out to observe the influence of salinity on seedling growth of germinating seeds
of canola, cabbage and cauliflower. The results indicated that an increased salinity concentration caused
delayed emergence of shoot and root compared to control. A continuous increase in length of shoot and
root was detected in frequent hours of germination in three plant species in the control as well as salt
treatments. The data on the average length (Table 2) of shoot and root shows that all species of Brassica
showed a strong inhibition with the increasing level of salt solution. There was considerable reduction in
the size of shoot and root at highest level of salinity (14.1 dS m-1 NaCl). These results showed sign of
great inhibition of shoot and root growth with NaCl treatments. The decrease in length of shoot was more
pronounced as compared to root in all NaCl salt treatments in canola, cabbage and cauliflower. However
this decrease was more prominent in canola then cauliflower and cabbage. Cabbage also showed sign of
great affects on shoot length. Great inhibition in root length was also recorded in cauliflower. In contrast,
the similar kind of inhibition of root growth was observed in canola and cabbage (Table 2).

Table 2. Effect of Different NaCl Treatments on the Length of (A) Root and Shoot (B) of Canola, Cabbage and
Cauliflower.

(A) Root length (cm) (B) Shoot length (cm)

NaCl (dS m-1) Cabbage Cauliflower Canola Cabbage Cauliflower Canola
0.00 5.97 3.87 5.40 4.20 2.37 9.20
4.70 5.40 2.37 4.50 2.20 2.17 5.00
9.40 4.67 1.93 4.33 1.87 1.90 3.37
14.10 2.53 0.70 2.30 1.03 0.80 2.06
LSD(0.05) 0.51 0.17 0.32 0.38 0.17 0.71

Highly significant differences were observed among the accessions for fresh weight of shoot and root.
The data presented in Table 3 revealed that fresh weight of shoot and root of all Brassica species was
reduced by increasing salt levels. Shoot and root fresh weight was significantly reduced in all accessions
at all salinity concentration, whereas fresh shoot weight was reduced more as compared to fresh root

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weight. This trend was more prominent in canola and cabbage then cauliflower at all salt levels (Table 3).
Salinity also strongly inhibited fresh root weight of all plant species. Cabbage showed more decrease
among all plant species. However canola showed less effect of salt solution at all salinity treatments in
case of fresh root weight (Table 3).

Table 3. Effect of Different NaCl Treatments on the Fresh Weight of (A) Root and Shoot (B) of Canola, Cabbage And
Cauliflower.

(A) Fresh root weight (mg) (B) Fresh Shoot weight (mg)
NaCl (dS m-1) Cabbage Cauliflower Canola Cabbage Cauliflower Canola
0.00 2.10 2.53 1.73 83.33 33.33 160.00
4.70 1.70 2.27 1.32 66.67 16.67 60.89
9.40 0.70 1.73 0.60 13.33 9.06 36.67
14.10 0.25 0.62 0.30 8.67 6.00 13.33
LSD(0.05) 0.21 0.23 0.21 6.78 9.64 12.72

Analysis of variance of the leaf area and number of leaves indicated that there were significant (P 0.05)
effects of NaCl stress on canola, cabbage and cauliflower (Table 4). Hence, the data presented below
indicated that leaf area of canola was comparatively more affected by salinity then cabbage and
cauliflower. Cabbage also shows sign of leaf area reduction as the salinity level increased but this trend
was little less in case of cauliflower (Table 4). Number of leaves was also significantly reduced as the
salinity concentration increased in case of all Brassica species. The reduction was little less in cauliflower
then rest of the plant species. Canola and cabbage almost showed similar response in case number of
leaves in all salinity treatments (Table 4).

Table 4. Effect of Different NaCl Treatments on the Leaf Area (A) and Number of Leaves Per Plant (B) of Canola,
Cabbage and Cauliflower.

(A) Fresh root weight (mg) (B) Fresh Shoot weight (mg)
NaCl (dS m-1) Cabbage Cauliflower Canola Cabbage Cauliflower Canola
0.00 8.09 1.03 20.41 6.00 4.33 6.00
4.70 6.14 0.49 4.89 4.67 3.33 4.67
9.40 5.10 0.33 3.54 4.33 2.33 4.33
14.10 2.66 0.18 1.87 2.00 2.00 2.33
LSD(0.05) 0.55 0.17 0.82 0.48 0.59 0.59

DISCUSSION

Final germination of all Brassica species decreased as the salinity level increased and salinity also
delayed germination rate (Table 1). It is assumed that germination rate and the final seed germination
decrease with the decrease of the water movement into the seeds during imbibitions [22]. Increasing
salinity concentrations in germination often cause osmotic and/or specific toxicity which may reduce or
retard germination percentage [23 and 24]. Salt induced inhibition of seed germination could be attributed
to osmotic stress or to specific ion toxicity [25]. Germination failures under saline soils are often the
results of high salt concentrations in the seed planting zone because of upward movement of soil solution
and subsequent evaporation at the soil surface [26]. Seed germination has been reported to decline with
effect of NaCl and incubation temperature increasing salinity levels [14]. These results are also similar in
line with Jamil & Rha [27]. They reported that germination of sugar beet and cabbage decreased as the
salinity concentration increased and salinity also delayed germination rate. Similar kind of results was
reported by Jeannette et al. [28]. They found that the mean time to germination of almost all Phaseolus
species increased with the addition of NaCl and this increase in median germination time was greater in
higher concentration as compared to low concentration. Some of the results obtained in this study are

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similar to those of some other workers who showed that in general, increased salinity results in decrease
in germinability and delayed rate of germination [29 and 30].

The shoot and root length are the most important parameters for salt stress because roots are in direct
contact with soil and absorb water from soil and shoot supply it to the rest of the plant. For this reason,
root and shoot length provides an important clue to the response of plants to salt stress [27]. It was
observed that the degree of reduction increased with the increasing concentration of salt in all Brassica
species (Table 2). The reason for reduced shoot and root development may be due to toxic effects of the
NaCl used as well as unbalanced nutrient uptake by the seedlings. High salinity may inhibit root and
shoot elongation due to slowing down the water uptake by the plant [31] may be another reason for this
decrease. Neumann [1] indicated that salinity can rapidly inhibit root growth and hence capacity of water
uptake and essential mineral nutrition from soil. Salt stress inhibited the growth of shoot more than root in
all Brassica species. Similar observations have been reported in barley (Hordeum vulgare L.) [25], pigeon
pea (Cajanus cajan) [32], tepary bean (Phaseolus acutifolius) [33] and tomato (Lycopersicon) [34].
Similar kinds of observations have been reported by Jamil and Rha [27] in sugar beet and cabbage that
salinity significantly reduced the root and shoot length at all salinity treatments

Fresh shoot and root weight of all plant species was strongly inhibited by all salinity treatments (Table 3).
Some researchers argue that the plants had the reduction in their fresh weights because of the proportional
increase in Na+ concentration, which could imply that an ionic effect was being manifested. However, one
could argue that because dry weights were not much affected compared to the fresh weights, growth
reduction would be attributable to osmotic effects. The results in this investigation are similar in line with
Shannon and Grieve [35]. They detected that Salinity reduced fresh weight (FW) of all nine vegetables.
Similar kind of result was observed by Jeannette et al. [28] that total fresh weight of root and shoot of
cultivated accessions was significantly reduced with increased salt stress.

Salinity had considerable effect on number of leaf per plant and also on leaf area in this investigation
(Table 4). Similar kind of results was obtained by Seema et al. [36] that varying levels of NaCl had
significantly reduced the number of leaves per plant in all plant cultivars. Salt stress reduces leaf growth
rate by shortening the length of the leaf elongating zone and decreasing the growth intensity in its central
and distal portions [37]. Leaf growth inhibition by salinity must be expected to occur via an effect on this
region [38]. Similar kind of response received by Grieve et al. [39] that leaf area decreased consistently
and significantly from a mean of 950 to 65 cm2 per plant as salinity increased from 3 to 21 dS m-1. Meena
et al. [40] also suggested that leaf area and relative water content decreased significantly with variable
magnitude with increasing salinity.

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