Effect of different salinity levels on maize (zea mays l ) genotypes by ITEFA DEGEFA

INT J CURR SCI 2016, 19(4): E 52-61 RESEARCH ARTICLE

ISSN 2250-1770

Effect of Different Salinity Levels on Maize (Zea mays L.) Genotypes During Germination Diriba Shankoa*, Itefa Degefab, Bula kerec and Gebayo Hayetod a,b c,Biology

Department, College of Natural and Computational Sciences (CNCS), Bule Hora University (BHU), P.O. Box: 144, Bule Hora, Oromia, Ethiopia dGraduate

student at Bule Hora University, Biology Department

*Corresponding author: diribashanko78@gmail.com Abstract Salinity is main threats to world food security, to ensure future food needs, production of salt tolerant crop varieties are necessary. This study was going to screen out the salt tolerant maize genotypes under salt stress condition. An experiment was performed as factorial under Completely Randomized Design (CRD) with three replications. The experimental treatments were contained four NaCl salinity levels (0, 80, 160 and 240 mM) and nine maize genotypes (61006, Mal-1, Mal.hybrid, 215065, MHQ, 207346, 227235, M-130 and 61036) were used in this experiment. Data was analyzed using SAS (version 16.0) statistical software and means were analyzed by least significance difference (LSD). Germination percentage (GP), Germination rate (GR), final germination percentage (FGP), radicle length (RL), plumule length (RL) and seed vigour (SV) were measured. The ANOVA for treatments, genotypes and their interaction was found to be highly significant (p<0.05) with regard to all parameters. Genotypes MHQ, 207346 and Mal.hybrid were salt tolerant genotype during germination but 227235 and 61036 genotypes were salt sensitive. The rest maize genotypes were intermediate in the salt stress condition. However, correlation analysis was demonstrated that positive correlation between GR and RL as well as between PL and RL, SV to salt stress. The results show that salt tolerant genotypes could be a better source for breeders for further evaluation on salt affected soil. This current study indicated that physiological indices are used as a selection tool for salinity tolerance in maize genotypes. Keywords: germination; NaCl; salinity; maize Received: 09th September 2016; Revised: 19thSeptember; Accepted: 22ndSeptember; ÂŠ IJCS New Liberty Group 2016 Introduction

sufficient

rainfall

which

percolates

salt

Crop productivity is seriously affected by soil

concentration from the root zone of crop plants that

salinity throughout the world. The deleterious effects of

mainly happened due to the presence of high

salt stress are obvious in soil, in arid and semiarid

evaporation rate (Almasouri et al., 2001). In addition,

climate owing to poor soil and water management

the existing of soil salinity problems in crop production

activities (Azevedo et al., 2006). In these areas, there is

will become worse due to rapidly growing human

Diriba et al., 2016

population from time to time in many countries and the

(Khodarahmpour, 2011). It is used as food for human

increasing over limited water resources which are

consumption as well as food grain for animals (Moussa,

forcing growers to use poor quality water for irrigation.

2001). Maize is the most salt-sensitive crop among

Approximately 800 million hectares of land in the

cereals crops (Ashraf and McNeally, 1990). Therefore,

world is affected by soil salinity. Seed germination and

this study aimed to screen salt tolerant maize genotypes

seedling growth are major physiological indices to

under various salinity levels during germination stage.

establish a vigorous crop stand which can be utilized to

Materials and Methods

screen crop plant germplasm for salt tolerance (Ashraf et al., 2007).

An experiment was conducted in a Biology laboratory at Bule Hora University Oromia, Ethiopia

In salt affected soil, deleterious effects of soluble

from March-May, 2015. Nine maize (Zea mays L.)

salts on plant development are due to decrease of

genotypes were used in this experiment such as 61006,

osmotic potential in plants that ultimately decreases

Mal-1, Mal.hybrid, 215065, MHQ, 207346, 227235,

water availability to plants, ionic imbalance and

M-130 and 61036. These genotypes were obtained

specific ion toxicity (Ashraf and Harris, 2004). Mainly

from Melkassa Agricultural Research Center and

it causes hyper-osmotic stress and hyper-ionic toxic

Institute of Biodiversity Conservation (IBC).

effects, which leads germination and seedling growth

NaCl Solutions and Seed Treatments

inhibition (Hasegawa et al., 2000). Salt reduces the

Seeds

were

sterilized

with

0.5%

sodium

ability of plants to utilize water and causes a reduction

hypochlorite (NaOCl) solution for 1 min (Sauer and

in growth rate, as well as changes in plant metabolic

Burroughs, 1986) at room temperature with 108 Petri

processes (Munns, 2002). So far, salt adversely affects

dishes. Petri dishes were washed and sterilized in hot

the plant growth and development, hindering seed

air oven at 700C for 36 hrs (Muhammad and Hussain,

germination (Dash and Panda, 2001). Seed germination

2010) and Whatman no.2 filter papers at 700C for 24

is usually the most critical stage in seedling

hrs. The petri dishes were lined with Whatman No. 2

establishment, determining successful crop production

filter paper and treated with 0, 80, 160 and 240 mM

and

2001;

concentrations of NaCl. The petri dishes were arranged

Bhattacharjee, 2008). It is a complex process which

in a Completely Randomized Design (CRD) in a

depends on the genetic and environmental factors, such

factorial combination with three replications (Gomez,

as temperature, light, and salinity (Mahmoud, 1985).

1984). Then, three seeds of each maize genotype for

So, salinity is one of the most important factors limiting

salt treatments were placed in a 90 mm uniform

plant growth and delaying seed germination as well as

distance Petri dish. The filter paper was moistened with

final germination percentage (Rahman et al., 2000).

distilled water for the controls. The Petri dishes were

Among cereal crops, maize (Zea mays L.) is the third

hermetically

most important crop in the world after wheat and rice,

evaporation (Asgharipour and Rafiei, 2011) and then

and grows under a wide range of climatic conditions

carefully kept in a humidity chamber at a temperature

productivity

(Almansourie

al.,

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sealed

with

parafilm

prevent

Diriba et al., 2016

of 25±1°C under 8hr/day length. Germination was

Radicle length

carried out in a germination chamber with 12hrs

The radicle length was measured from the point of

photoperiod. The numbers of germinated seeds were

first cotyledonary node to the tip of longest root and

counted every day for 12 days from the start of the

expressed in cm plant-1.

experiment. Treatment application with the same

Statistical analysis

amount of salt solution was continued every other day

Data analysis was carried out using SAS (version

and germination count started after 6 days of sowing

16.0) statistical software (SAS Institute Inc., USA)

and continued until the 12th day. The seeds were

where two way analysis of variance (ANOVA) and

considered to have germinated when both the plumule

correlation analysis was done. Whenever means of

and radicle have been emerged greater than 0.5 cm.

treatment differences were significantly at p<0.05,

Germination Percentage The germinated seeds were

which is separated by using the Least Significant

counted daily according to the seedling evaluation

Difference (LSD) test (Duzgunes et al., 1983).

procedure described in the Handbook of Association of

Results

Official Seed Analysts. The number of germinated

Analysis of variance showed that there were

seeds was recorded every 24 hrs (AOSA, 1990).

significant differences between salinity stress levels.

Twelve days after germination, the germination

The results of this study revealed that various

percentage (GP) was obtained by dividing the number

concentrations of NaCl had a significant effect on the

of germinated seeds in any petri dishes by the total

all measured traits for maize genotypes. And also,

number of seeds, multiplied by 100 (Cokkizgin and

analysis of variance showed that interaction effects

Cokkizgin, 2010; Tanveer et al., 2010).

were significant for all investigated traits (Table 1).

Germination rate

Germination Rate

Germination rate (GR) which is the average

Analysis of variance exhibits significantly reduced

number of days needed for plumule or radicle

seed germination rate in all maize genotypes (Fig. 1a)

emergence was calculated using MAGUIRE’s equation

which resulted significant at (p≤0.05) variations in their

(Maguire 1962) as: GR (M) = n2/t2 + n4/t4 + n6/t6… +

interaction with respect to germination rate. The results

n12/t12; where n2, n4, n6 .., n12 represent the number

of this experiment indicated that genotype 227235,

of germinated seeds at times t2, t4, t6 …, t12 (in days).

Mal.hybrid and 207346 followed by M-130 and 61036

Plumule length

gave significantly higher mean germination rate than

Plant height was recorded by measuring the height

the other genotypes in all salinity levels. Genotypes

of the seedling from the surface of the soil to the tip of

207346,

MHQ,

the early seedling. This was recorded on 12th day after

significantly faster germination rate than the other

treatment and expressed in cm plant-1.

genotypes were tested in the 240 mM treatments.

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mal.hybrid

and

61006

showed

Diriba et al., 2016

Germination percentage

is important for initiation of any breeding program

The differences between the means of genotype

because it provides a chance for selection of desirable

(Fig. 1b) and salinity stress levels were compared by

genotypes with desirable traits simultaneously (Ali

Duncan multiple range test. It was observed that in all

et al., 2009). Correlation coefficient between all

of genotypes, there was a decrease in germination

possible combinations was estimated and is shown in

percentage. Though, in this experiment different maize

the table 2.

genotypes had different response to the salinity stress.

Discussion

Among the maize genotypes, MHQ and 207346 had the

The main purpose of this study was to identify salt

highest mean germination percentage at 160 and 240

tolerant genotypes in maize germplasm at germination

Mm NaCl levels. However, maximum reductions in

stages under different salinity levels. The analysis of

germination percentage of certain maize genotypes

variance for genotypes, NaCl levels and their

were observed at 240 mM of NaCl.

interaction was found to be significant for all

Radicle length

parameters; reflecting that all the genotypes responded

As it was clearly determined that there were

differently to salt stress with respect to all parameters.

significant differences between studied genotypes at

Increment in NaCl salinity level caused significant

salinity levels 80, 160 and 240 mM for the length of

reduction in germination rate (GR) of maize genotypes.

radicle. Among the maize genotypes, Mal. hybrid,

However, the reduction was sharp at 80, 160 and 240

MHQ and M-130 were scored the highest mean of

mM NaCl salinity levels. On the basis of germination

radicle length at 80 mM (Fig. 1d).

rate for the soils having 240 mM salinity, the genotypes

Seed Vigour

207346, MHQ and 61006 can be recommended

Seed

vigour

decreased

with

increase

(Figure.1a). An effect of salt stress on seed germination

concentration of NaCl solution. The best level of seed

rate might be the same which has been reported in

vigour was achieved at control treatment of NaCl

previous studies (Ashraf and Abu-Shakra 1978). There

concentration (Fig. 1e). From all genotypes, MHQ

are many reports which indicate that the genotypes

produced highest seed vigour at all salinity levels.

which maintained higher germination rate under saline

Correlation analysis

conditions produced higher biomass and yield (Ashraf

Breeders always look for genetic variation among

et al., 2006; Krishnamurthy et al., 2007). Also, many

traits to select desirable types. Some of these traits are

results showed that increased salt concentration reduces

highly associated among themselves and with seed

germination rate, in several maize genotypes (Cicek

yield. The analysis of the relationship among these

and Cakirlar, 2002). The reduction in germination rate

traits and their association with seed yield is essential to

is due to toxic effects of certain ions and also higher

establish selection criteria for salt tolerant genotype

concentration of salt reduces the water potential in the

(Singh et al., 1990). And also, the information

medium

regarding significant correlation among the characters

germinating seeds and thus reduces germination (Jamil

which

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hinders

water

absorption

Diriba et al., 2016

et al., 2006). Higher salinity level retard seed

maintaining their proper nutritional requirements

germination and radicle emergence due to osmotic

necessary for their healthy growth (Krishnamurthy

effect which is deleterious and prevent the plants from

et al., 2007).

Table 1. Analysis of variance on Mean Squares of measured traits of maize genotypes under salinity stress Source of variance

RAL

11766.9**

1.27**

1632.79**

6.98**

44.72**

82473.48**

Varieties 8 721.7** 0.02** 273.78** SL x varieties 24 184.1** 0.01** 487.22* Error 35 45.49 0 187.81 *, **Significant at 5% and 1% probability levels, respectively

0.36** 0.09* 0.02

0.65** 0.25* 0.21

76548.03** 46753.28** 4208.67

Fig. 1 (a-e). Effects of different salinity levels on Germination rate, Germination percentage, Final Germination Rate, Radicle length and Seed vigour of nine maize (Zea mays L.) Genotypes

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Diriba et al., 2016

Table 2. Correlation Coefficients of studied Traits of Maize Genotypes under Different Salinity Levels GR FG PL GP 0.887** -0.421* 0.875** GR 1 -0.395* 0.852** FG 1 -0.44** PL 1 RL SV *, ** Significant at 0.05 and 0.01 probability levels, respectively,

RL 0.916** 0.927** -0.45** 0.902** 1

SV 0.882** 0.748** -0.385* 0.725** 0.747** 1 GP= Germination percentage, GR=Germination

rate, FG = Final germination, PL= Plumule length, RAL= Radicle length and SV= Seed vigor

was

replacement of Ca2+ with Na+ and leakage of K+ (Takel,

obtained at zero salinity level (Fig.1b). The results of

2000). The length of radicle is one of the most

this experiment also agreed with the results of Farsiani

important traits for salinity stress because roots are in

and Ghobadi (2009) and Khayatnezhad et al. (2010) in

contact with soil and absorb water from soil. For this

maize. This results accordance, with Rejili et al. (2010)

reason, the length of radicle provides an important clue

observed a decrease in germination percentage that

to the response of plants to salinity stress. A marked

resulted due to osmotic effect of salts present in the

reduction in the length of radicle, the length of plumule

growth medium. Salinity causes a decrease in radicle

and the growth of seedling of all genotypes of maize

and plumule growth, which increased with the increase

was observed due to salt stress. Among the maize

in salinity levels. Presence of salinity in the growth

genotype, MHQ, Mal.hybrid, Mal-1 and 207346 had

medium reduced the absorption of water due to

the highest length of radicle, while, 61036 had the

decrease in osmotic potential of the medium which

lowest length of radicle (Figure.1d). Maize genotype

adversely affects cell division and differentiation

MHQ and 207346 also had the highest length of

(Ashraf et al., 2005). This experiment also explains, the

plumule (Fig. 1c). Results of this study showed that

length of plumule and radicle are negatively affected by

length of radicle and plumule decreased with increasing

salinity. According to Kausar et al. (2012), the length

salinity levels in all genotypes. Also, it was clearly

of plumule and radicle retardation is due to salinity

determined

effect and other reports also may be due to ionic

differences between studied genotype at salinity level

toxicity, disturbance in nutrients uptake (Akhtar et al.,

240 mM for the length of radical and plumule. The

2012), osmotic effects of salinity (Ashraf et al., 2005)

most effective levels in reducing these attributes were

and water absorption (Ashraf and Sarwar, 2002),

160 mM of NaCl. Best level of NaCl concentration in

resulting in reduction in biosynthesis of enzymes, plant

radicle length and plumule length was at control

hormones necessary for seedling growth (Bor et al.,

treatment. This results are in agreement with many

2003). The salinity stress imposed at germination stage,

researches (Mohammadkhani and Heidari, 2008;

damages cell membranes of the seedling due to which

Farsiani and Ghobadi, 2009; Jajarmi, 2009; Gholamin

cell membrane permeability increases, resulting in the

and Khayatnezhad, 2010; Khayatnezhad et al., 2010).

The

maximum

germination

percentage

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that

there

were

steadily

significant

Diriba et al., 2016

Among the maize genotypes, 227235, M-130 and

each other, except the GP with GR, FG with PL, RL

61036 were highly affected by salinity stress at 160 and

and SV coefficients.

240mM salt levels; because it gave the highest

Acknowledgements

reduction rate for seed vigour (Figure.1e). The results

Our gratitude and thanks to Biology department

was agreed with the work studied by Mostafavi et al.

staffs for their unreserved help and assistance during

(2011) which showed that four maize varieties in

laboratory and field work. We would like to express

drought and salt stress conditions reported that hybrid

our heartfelt thanks to Bule Hora University for its

KSC704 produced the highest seed vigour. The results

financial assistance and for providing the research site

of correlation showed that germination percentage (GP)

and also the Melkassa Agricultural Research Center

had the most positive and significant correlation with

(MARC) and Institute of Biodiversity Conservation

radicle length (r=0.92**). Germination percentage (GP)

(IBC) for supplying all the maize genotypeâ&#x20AC;&#x2122;s seeds.

depicted a negative and significant correlation with

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