Effect of Salt (NaCl) Stress on Germination and Early Seedling Growth

8/13/2019 Effect of Salt (NaCl) Stress on Germination and Early Seedling Growth

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Annals of Biological Research, 2011, 2 (4) :490-497

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ISSN 0976-1233

CODEN (USA): ABRNBW


Effect of salt (NaCl) stress on germination and early seedling growth

of Spinach (Spinacia oleracea L.)

1Mohammad Hosein Bijeh keshavarzi*, 2Mehrnaz S. Ohadi Rafsanjani, 1S. Mohsen.Moussavinik and

3Amir Parviz Lak

1Department of Agronomy, Zabol University, Iran

2Dept. Biotechnology, F/O- Science, Jamia Hamdard, New Delhi, India, India

3M.Sc. Post graduated in industrial management, Islamic Azad University, Tabriz Branch

______________________________________________________________________________

ABSTRACT

Salinity is one of the major environmental factors that lead to a deterioration of agricultural

land and reduction in crop productivity worldwide. This research was carried out in order to test

the effects of different salinity levels on germination and early growth of Spinach seedlings. The

experiment was carried out using completely randomized design in four replication in 2011 at

Hamdard University laboratory in India. Experimental treatment includes 4 levels of NaCl

concentration (o, 50, 100 and 150 mM). Result showed that the percentage and speed of

germination, plumule length, radicle length and heaviest wet and dry seedling weights were

higher in control treatment. At 150 mM and more concentration, germination decreased

significantly. This reduction in germination indicates this plants extreme insensitivity to salinity,

so it isnt advisable to cultivate it in saline soil. All the result data analyzed by SAS software and

comparison of means had been done with Duncan test in 0.05% probable level.

Key words: germination, NaCl, salinity stress, seedling, Spinacia oleracea L.

______________________________________________________________________________

INTRODUCTION

More than 900 million hectares of land world-wide, approx. 20 % of the total agricultural land,

are affected by salt, accounting for more than 6% of the worlds total land area. NaCl is the

predominant salt causing salinization, and it is unsurprising that plants have evolved mechanisms

to regulate its accumulation [1].


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M. H. Bijeh keshavarziet al Annals of Biological Research, 2011, 2 (4):490-497_____________________________________________________________________________


Seed germination is an important and vulnerable stage in the life cycle of terrestrial angiosperms

and determines seedling establishment and plant growth. Despite the importance of seed

germination under salt stress [2], the mechanism (s) of salt tolerance in seeds is relatively poorly

understood, especially when compared with the amount of information currently available about

salt tolerance physiology and biochemistry in vegetative plants [3, 4, 5, 6]. In vegetative plants,

salt stress causes reduced cell turgor and depressed rates of root and leaf elongation [7, 8],suggesting that environmental salinity acts primarily on water uptake. Furthermore, high

intracellular concentrations of both Na+ and Cl

- can inhibit the metabolism of dividing and

expanding cells [9], retarding germination and even leading to seed death.

The different results were dedicated from the effect of salinity stress on the quantitative and

qualitative parameters. For instance, it was found that increasing of salinity stress decreased

almost all of growth parameters in Nigella sativa some growth parameters and essential oil

amount in chamomile [10]. Also Younis et al. [11] reported that enhancing salinity treatments

lead to growth reduction. It also reduces germination amounts and seedling weight. Ashraf and

Orooj [12] reported that salinity treatment lead to reduction of growth and plant developments.

Overall, salinity through enhancement of osmotic pressure leads reduction of water absorbanceand disturbance in metabolic and physiological processes will be under its effect. So it cause

more delay in germination following by enhancing seed germination duration [13].

Spinach (Spinacia oleracea L.) is a vegetable and belongs to the family Amaranthaceae. It is

native to central and southwestern Asia. Spinach has a high nutritional value and is extremely

rich in antioxidants, especially when fresh, steamed, or quickly boiled. It is a rich source of

vitamin A (and especially high in lutein), vitamin C, vitamin E, vitamin K, magnesium,

manganese, folate, betaine, iron, vitamin B2, calcium, potassium, vitamin B6, folic acid, copper,

protein, phosphorus, zinc, niacin, selenium and omega-3 fatty acids. Recently, opioid peptides

called rubiscolins have also been found in spinach. Polyglutamyl folate (Vitamin B9 or folic

acid) is a vital constituent of cells and spinach is a good source of folic acid, but boiling spinach

can more than half the level of folate left in the spinach, though microwaving does not affect

folate content [14].

MATERIALS AND METHODS

The experiment was carried out using completely randomized design in four replication and 4

salinity levels (0, 50, 100 and 150 mM) in 2011 at Hamdard University laboratory in India. Each

experimental unit includes 1 Petri dish with 100 150 mm dimension each contains 15 healthy

and homogenous seeds which were put on the No1 Watman filter paper. First of all, to disinfect

seeds, we put them in 10% Hypochlorite Sodium solvent then we washed them 3 times bydistilled water. Next, we added 6 ml NaCl solvent to each Petri dish in this way filter water was

weltered by NaCl completely. Eventually, their lids were closed by parafilm and had been

located in growth room. The temperature adjusted in 25oC. This experiment took 7 days.

The following characteristics were studied:

Germination Percentage (GP):From second day, the germinated seeds were counted daily in specific time. At that time, those

seeds were considered germinated which their radical length was more than 3 mm.

Counting continued till we could count more germinated seeds and the resulted final counting

considered as final germination percentage.


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GP: Ni / N 100

Ni: number of germinated seed till ith

day

N= total number of seeds.

Germination Race (GR):In order that, from the second day to 7

thonce in 24 hours we counted germinated seeds and its

race was determined by Maguire equation [15]:

GR: Germination Race (number of germinated seed in each day)

Si: number of germinated seeds in each numeration

Di: number of days till nth

numeration.

n: number of numeration times.

Seed vigor (SV):This index was determined by the following formula and with the help of Abdul-baki and

Anderson [16] method:

Strong seed index = {germination percentage means of seedling length (radicle + plumule)

mm} / 100

At the end of experiment, we had selected 10 plants from each Petri dish, separated their radicle

and plumule and measure each plats radicle and plumule length separately. Then we put each

repetition on the filter separately. In order to make them dry and measure its dry weight, we putthem in oven with 75

oC temperature for 24 hours. Thereafter, we weighed them and data was

analyzed using SAS and Excel software.

RESULTS AND DISCUSSION

Table 1: result of variance analysis on 7 seed germination and growth of seedling characteristics under NaCl concentration

Mean Square

S.O.V df GP (%) GR RL (cm) PL (cm) Seed vigor WW (g) DW (g)

Replication 3 30.73ns 0.64ns 0.055ns 0.058ns 0.18ns 0.0003ns 0.000091ns

Treatment 3 2634.9** 48.92** 13.17** 4.15** 34.2** 0.086** 0.0045**

Error 12 27.6 0.45 0.07 0.048 0.14 0.00026 0.000057

C.V. (%) 8.37 8.86 8.13 11.74 9.71% 6.25 14.85Note: *and ** indicate significant difference at 5% and 1% probability level, respectively ns is not significant.

GR: Germination rate, GP: Germination percentage, PL: Plumule length, RL: Radicle length, SV: Seed vigor, WW: Wet weight,

DW: Dry weight.

Table 2: Effect of different NaCl concentration on seed germination and growth of seedling characteristics

NaCl concentration (mM) GP (%) GR RL (cm) PL (cm) SV WW (g) DW (g)

0 86.25a 10.32a 5.52a 3.15a 7.47a 0.41a 0.086a

50 72.5b 9.31a 3.72b 2b 4.14b 0.308b 0.057b

100 61.25c 6.04b 3.1c 1.35c 2.72c 0.183c 0.03c

150 26.25d 2.6c 1.12d 0.77d 0.5d 0.07d 0.0087d

Note:Similar letters in each column hadnt any significant statistical difference.; GR: Germination rate, GP: Germination

percentage, PL: Plumule length, RL: Radicle length, SV: Seed vigor, WW: Wet weight, DW: Dry weight.


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Results of statistical analysis of experimental data have been given in table 1 and results of

comparison between considered characteristics means have been written in table 2. As table 1

show, salinity made significant differences on all considered characteristics.

Germination percentage and race:

According to results of variance analysis, effect of salinity stress level on germination percentageand race were significant (P < 0.01) (Table 1). Comparison between means of different level of

salinitys effects on germination race and percentage has been showed in table 2. As you see in

salinity stress, the most germination percentage was high (86.25%) and reduced at 150mM NaCl

concentration. (26.25%) (Figure1).

Figure 1: Effect of different levels of NaCl on germination percentage of Spinacia oleracea L.

The highest germination race was related to control also with (10.32), while lowest with 150 mM

with (2.6) (Figure 2). Its cause could be more than usual presence of anion, cation which in

addition to toxication, decreased water potential that is because of its solubability in water. Ions

so plant cant absorb water and encount to lake of water [17]. We also can say that this reduction

in germination race relies on salinity could be because of its bad effect on physiological

processes which are effective on seed germination [18].

Figure 2: Effect of different levels of NaCl on germination rate of Spinacia oleracea L.


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Radicle and plumule length:The effects of salinity stress on radicle and plumule length have been showed in table 2 Results a

significant difference in radicle and plumule length in 0.05% probable level (Table 1).

Comparison of radicle and plumule length means in salinity different level (0, 50, 100 and 150

mM) showed that when salinity level increase, seedlings radicle and plumule length decrease.

The most reduction in radicle length (Figure 3) and plumule (Figure 4) related to 150 mM. In thisrelation Munns and Termaat [19] suggested that salinity decrease radicle and plumule growth

and if we increase salinity level, the amount of reduction will increase. Also Salinity, declines

plumule and radicle growth, and by increasing salinity these reduction increase. Salinity which is

result of osmotic pressure leads reduction in water absorbance so cell division and differentiation

reduce and reduction of plumule and radicle length will be Explainable.

Figure 3: Effect of different levels of NaCl on radicle length of Spinacia oleracea L.

Salinity causes shorter plumule, which is more in case of NaCl more than other salinity factors

gas deterrent impact on embryo tissues appearance [20].

Figure 4: Effect of different levels of NaCl on plumule length of Spinacia oleracea L.

In addition, Hajar et al. [21] by studdingNigella sativa L.different salinity treatment till 300 mM

NaCl. They conclude that, in Nigella sativa L.root growth will decrease if salinity increase till

150 mM NaCl. Some studies showed that germinated seeds in salinity environments have short


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root and shoot and NaCl, has on extreme deterrence effect on embryo development rather than

other salinity factors [22, 23].

Seed vigor:In strong seed vigor index, had been observed that there exists a significant difference (P 0.01)

between different salinity levels (table 1). By increasing NaCl concentration, seed vigor indexdeclines (Table 2, Figure 5). The most seed vigor index was related to control treatment (7.47)

and the least at was related to 150 mM (0.5) (Figure 5).

Generally, race and percentage of germination and seed vigor index is related to special impact

of ions and reduction of environmental water potential in the presence of salinity. Result showed

that if salinity increases (reduction of environmental osmotic potential), seed characteristics will

decrease these results are in accordance with the founding of Kader and Jutzi [24].

Figure 5: Effect of different levels of NaCl on seed vigor of Spinacia oleracea L.

Figure 6: Effect of different levels of NaCl on dry and wet weight of Spinacia oleracea L.

Wet and dry weight:Impact of salinity stress treatments, on dry and wet weight of Spinacia oleracea L.seedling was

significant (P 0.01) (Table 1). Impact of salinity stress on dry and wet weight had been showed

in Table 2. As you see by enhancing salinity levels, seedlings wet weight amounts decrease

extremely. In this case in 150 mM we have 0.07 gr also in other treatments (100, 50 and 0 mM),


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dry weights were 0.183, 0.308 and 0.41 in orderly. (Table 2, Figure 6). In addition, dry weight of

seedling have similar results which when we increase salinity level till 150 mM dry weight

decreases, means that it decrease from 0.086 gr to 0.0087 gr. (Table 2, Figure 6).

Etesami and Galeshi [25] reported that salinity is the cause of reduction in germination

percentage, race and homogeneity of germination and dry weight of barley (Hordeum vulgare)seedling. Massai et al. [26] say that salinity is delaying plant growth under reduction of

photosynthesis effects, it is cause of closing stomata and reduction of water entrance into the

plant and so that it cause duplicate reduction in plant weight.

Redman et al. [27] showed that this reduction in dry weight of plumule and radicle which is

results of enhancing the salinity concentration is a normal phenomenon and probably it is the

result of low water absorbance by germinating seeds.

ACKNOWLEDGMENT

Authors are thankful to M.Z. Abdin, Head, CTPD, Dept. of Biotechnology, Jamia HamdardUniversity, New Delhi, India for providing laboratory facilities

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Effect of Salt (NaCl) Stress on Germination and Early Seedling Growth