沙漠研究 24-1, 183-186 (2014)
Ἃ₍◊✲ 24-1, 183 - 186 (2014)
Journal of Arid Land Studies
Journal of Arid Land Studies－ DT11 Refereed Paper －㸫DT11 Refereed Paper㸫

Salinity Tolerance Evaluation of Asian Barley Accessions (Hordeum vulgare L.)at the Early Vegetative Stage
Hanen SBEI1), Tariq SHEHZAD1, 2), Moncef HARRABI3) and Kazutoshi OKUNO*1, 2)
Abstract: Salinity is one of the major constraints on world barley production. Exploitation of natural variation of salinity tolerance
in existing germplasm and the development of tolerant cultivars are potentially effective approaches for minimizing yield losses. In
this study, seedlings of 146 accessions were assessed in a hydroponic system. The experiment was arranged in a completelyrandomized design with the treatment of 0 and 22 ms/cm2 NaCl, three replications and four plants in each accession per replication.
After 17 days of treatment, salinity tolerance index (STI) was calculated for the traits: number of leaves (NL), shoot length (SL), rootlength (RL), shoot dry weight (SDW), root dry weight (RDW) and plant dry weight (PDW). There was a wide variation in salinity
tolerance among the barley accessions. All traits were affected by salinity. SDW was highly affected by salinity and correlated with
the NL, RDW and PDW, with the correlation coefficients of 0.43, 0.84 and 0.99, respectively. Based on a multivariate analysis, 19
tolerant and 14 susceptible accessions were identified.
Key Words: Barley, Multivariate analysis, Salinity tolerance, Seedling

1. Introduction
Salinity stress is a major constraint to agricultural foodproduction because it decreases crop yield and restricts the useof agricultural land. It was estimated that salinity affectednearly 950 million ha (6%) in the world (Babu et al., 2007).
Approximately 20% of agricultural land in the world andnearly half of all irrigated land suffer from salinity (Flowersand Yeo, 1995; Munns, 2005). The area affected by salinity
in South, South East, North and Central Asia covered 25% of
the total soil. As a result, the development of salinity-tolerant
crops is an important option for maintaining crop production insaline soil.
Barley (Hordeum vulgare L.) is the fourth most importantcereal crop worldwide and is an important source for feed, malt,and human food. Its importance is derived from the ability togrow and produce in marginal environments, such as drought,
low temperature and salinity (Hayes et al., 2003). The
genetic diversification and the adaptability to a broad range of
ecological conditions have highly strengthened the salinity
tolerance in barley. These factors might have raised a richgene pool with a large variation in adaptation to salinity.
Salinity tolerance of barley germplasm collections wasevaluated by different parameters including relative watercontent, germination rate, coleoptile length, stem and radicle
length, as well as dry and fresh weight of roots and shoots* Corresponding Author: okuno.kazutoshi.fu@u.tsukuba.ac.jp(Mano and Takeda, 1997; Xue et al., 2009). The screening
was done at germination and the vegetative stage according toprevious reports (Mano and Takeda, 1997-1998; Ellis et al.,
2002; Farhad and Mohammad, 2008) and at late stage in fieldby Xue et al. (2009).
The primary objectives of this work were to 1) investigate
the genetic variation in salinity tolerance of Asian barleyaccessions at the early vegetative stage and 2) select the highly
tolerant and susceptible accessions using the multivariate
analysis.

2. Methods and Materials
2.1. Materials and hydroponic culture
A total of 146 barley accessions from six Asian countries
were used in this study: Bhutan, China, India, Japan, Korea andNepal (Table 1). Barley accessions were provided by
Institute of Plant Science and Resources, Okayama University
with support in part by the National Bio-Resource Project ofthe MEXT, Japan.
The accessions were screened for salinity tolerance atseedling stage in a growth chamber with natural light, and24/20°C day/night temperature. The experiment was laid out
in a randomized design with three plots for untreated and 22.04
ms/cm2 of NaCl (250 mM). A total of 12 seedlings peraccession were evaluated (4 seedlings per treatment).
Uniform seedlings were selected based on the evaluation of㸦Received, September 15th, 2013; Accepted, January 4th, 2014㸧

1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan
1) Graduate School of Life and Environmental Sciences, University of Tsukuba, Japan2) The Alliance for Research on North Africa, University of Tsukuba, Japan

3) Plant breeding Laboratory, National Institute of Agriculture at Tunisia

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Table 1. Asian accessions used in this study.

SLRLSDW

145

238.6** 266**

432**

16NS

41**

15**

22901384NS 2163** 619**
1415** 51**
51**

26 NS
15NS

93*
32*

21NS
11**

145

1080** 174**

99**

386**

21**

7**

22903995** 142**
1991** 50**

797**
25**

438**
264**

29*
19**

19**
7**

2.2. Salinity Tolerance Evaluation
After 17 days of salinity treatment, seedlings were
harvested and root length (RL) and shoot length (SL) wererecorded. Then, plants were separated into roots and shoots,
dried in an oven at 85°C for 72 hours and weighed. Salinity
tolerance index (STI) was estimated according to the relative
ratio of trait under treated condition by control based on shootand root data.

3. Results and Discussion
3.1. Salinity tolerance evaluation
As shown in Table 2, there was a significant variation
among accessions in all traits under control conditions except
the SDW. No significant variation among replications wasBA


Number of accessions
Number of accessionsCD

Number of accessions

Number of accessions

EF

Number of accessions

2.3. Statistical analysisThe growth parameters, such as shoot and root dry weight,
the GLM analysis, the principal component analysis and thecluster analysis were subjected to statistical analysis using
software JMP V10 software, ver. 9 (SAS Institute Inc., 2010).

PDW

Acc: Accession; R: Replication; NS: Not significant; ** and *:Significance at 5% and 1%, respectively.

146

root length, shoot length and root aspect, then transplanted into
containers filled with hydroponic solution of the following
proportion (%): total N (10%), P2O5 (8%), K2O (27%), MgO(4.0%), MnO (0.10%), B2O3 (0.1%), CaO (23%), Fe (0.18%)
with Cu, Zn and Mo traces. The salinity treatment wasstarted when two leaves were completely developed. To
reduce the effect of osmotic shock, initial salinity concentration
was 13.81 ms/cm2 and then increased within 3 days to reachthe final concentration 22.04 ms/cm2 of NaCl. Then, thetreatment was maintained for 14 days. The solution waschanged every week. The pH of solution was maintained at
6.5.

RDW

Frequency (%)

Nepal
Total

Landrace
Landrace

Acc*RNL

Frequency (%)

Korea

20102030Acc*R
S ALT
Acc
Rddl

Frequency (%)

Japan

Improved
Landrace
Improved
Landrace
Improved

2541010VSControlAcc
R

Frequency (%)

India

Number of accessions
710Frequency (%)

China

Categories
Landrace
Improved
Landrace

Frequency (%)

OriginBhutanTable 2. GLM statistical analysis (Mean square) for Asian barley
under control and salt treatment.

Number of accessions

Fig. 1. The frequency distribution of the STI for the traitsnumberof leaves (A), shoot length (B), root length (C), shoot (D), root(E) and plant (F) dry weight.

observed for the NL, SDW and PDW. SDW was not affectedby the environment changes (Acc* R) and within replications.
Under salt treatment, a significant variation among theaccessions, replications and interactions was observed for alltraits.
There was a wide difference in the response to salinity
stress among the 146 accessions indicating a great difference insalinity tolerance among Asian landraces and improved
cultivars (Fig. 1). On the average, NL, SL, RL, SDW, RDW

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Table 3. Mean, standard deviation, coefficient of variance and 95%
upper and lower confidence interval for STI of all traits.

STINL
STISL

66
68.7

11.4
8.1

CV(%)
17.3
11.8

STIRL
STISDW

79.7
55.1

16.5
19.6

20.7
35.8

STIRDW
STIPDW

70.1
58.7

20.2
19.5

28.8
33.3

STIM ean

Sdtv

Upper%

Lower%

Range%

67.9
70
82.5

64.1
67.3
78.1

35- 9550- 9550-140

58.3
73.5
62

51.8
66.8
55.5

30-107
20-117
20-107

Fig. 2. Principal component analysis for Asian barley accessions
according to the salt tolerance index (+: Bhutan; °: India; ‫ݡ‬:China; ‫ۂ‬: Japan; ന: Korea ‫ڦ‬: Nepal).

and PDW of the treated plants were 65, 68, 79, 55, 70 and 58%of the control. The standard deviation ranged from 8 to 20%.
The SDW coefficient variation was the highest comparedwith the other traits with a CV = 35.8%. This indicates thatSDW is more affected than others morphological traits whenplants are exposed to salinity stress in hydroponic culture at anearly stage (Table 3). In addition, SDW was correlated to theNL, RDW and PDW with the correlation coefficients of 0.43,0.84 and 0.99, respectively.
3.2. Multivariate analysis
3.2.1. Principal component analysis
The principal component analysis (PCA) based on the STIof all traits was used for discrimination between accessions(Fig. 2). PCA represented 75% of variability. The biomass(SDW, RDW and PDW) explained 55.1% of the variability
whereas component 2 explained 20% of the variability andwas influenced by SL and RL. The accessions originatedfrom Bhutan and Japan maintained the highest biomass
production compared to others accessions. The Nepalesefollowed by the Indian accessions are the least productive
under salt treatment. The Korean and Chinese accessions wereFig. 3. Cluster analysis using STI for all the traits of the 146 Asianaccessions.

classed as moderately tolerant to salinity.
3.2.2. Cluster analysis
Cluster group rankings were obtained based on average
linkage (between groups) method using the squared Euclidian
distance on the means of the salinity tolerance indexes for all
traits at early vegetative stage (Fig. 3). The rescaled distancecluster ranged from 1 to 25.
The accessions were divided into two groups consisting of
tolerant accessions and both moderately tolerant and highly
susceptible at the similarity level 18. The most tolerantaccessions were the landrace from Bhutan, improved cultivars
from China and landraces from Japan. Chinese and Koreanlandraces with the improved cultivars from Japan were ranked
as moderately tolerant accessions. The most susceptible
accessions were the Indian and Nepalese landraces. By

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multivariate analysis, 19 susceptible and 14 tolerant accessions
were identified and used for further research.
3.3. Discussion
Screening for salinity is the basic and primordial step forplant breeding needed to make crosses between susceptible
and tolerant accessions, to get inbred lines, and to identify thequantitative traits loci responsible for salinity tolerance.
Several previous studies have been reported (Eleuch et al.,2008; Qui et al., 2011; Wu et al., 2011; Sbei et al., 2012; Longet al., 2013) that the variation in salt tolerance exists among
European, African and Asian Hordeum vulgare. Ellis et al.
(2002) used a variety of barley germplasm accessions from theMiddle East for breeding. In the current study, East Asian
accessions were screened for salinity tolerance under 22 ms/cmof NaCl in hydroponic culture. The results revealed a widevariation in salinity tolerance among the accessions. In
particular, most of the accessions originated from Nepal werehighly susceptible, whereas Japanese and Bhutan landraces
were highly tolerant to salinity. The difference in response tosalinity stress among accessions could offer elite alleles
conferring salinity tolerance.

4. Conclusion
The variation in salinity tolerance in Asian barleyaccessions could be exploited and used for the barley breeding
program. Tolerant and susceptible accessions were selected,
according to the STI of each trait, for further study on detectionof QTL associated with salinity tolerance.

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