SALINITY AND ITS EFFECTS ON THE PHYSIOLOGICAL RESPONSE OF BEAN.pdf

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ORIGINAL PAPER

749Volume 9 (2008) No. 4 (749-756)

SALINITY AND ITS EFFECTS ON THE PHYSIOLOGICAL RESPONSE OF BEAN

(PHASEOLUS VULGARIS L.)

(PHASEOLUS VULGARIS L.)Miroslava Kaymakanova, Nevena Stoeva*, Tsvetana Mincheva

*Agricultural University, 12, Mendeleev Str, 4000 Plovdiv Bulgaria, e-mail: [email protected]

Manuscript received: March 18 2008; Reviewed: July 9 2008; Accepted for publication: February 11 200

ABSTRACT

T e e ect o sa t stress n t e p ys o og ca react on n young ean p ants was stu e . T e p ants were grown n pots

as hydroponic cultures in half-strength Hoagland nutrient solution under controlled conditions in a climatic room. The

plants were treated for 7 days with NaCl and Na SO (concentration 100 mM),starting at the appearance of the first

trifoliate leaf unfolded. The salts were added to the nutrient solution.It was esta s e t at t e equ mo ar concentrat ons o ot sa t types cause stress n t e young ean p ants,

w c oun express on n t e suppress on o growt , p otosynt es s act v ty an cause c anges n stomata status

(conductivity, number and size). The transpiration and the cell water potential in salt-treated plants were reduced. The

MDA level in root and shoot, and the proline content was increased.

Key words: bean, salinity, salt stress, growth, leaf-gas exchange, water potential, stomata number and size.

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NaC Na SO 100 mM ,. .

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50ournal of Central European Agriculture Vol 9 (2008) No 4

iroslava Kaymakanova, Nevena Stoeva, Tsvetana Mincheva

INTRODUCTION

he salinity of soil is a widespread environmental problem

an mportant actor n m t ng agr cu tura pro uct v ty.

Sa n ty a ects p ants t roug osmot c e ects, on-

spec c e ects an ox at ve stress 19 . T e g sa n ty

evels of soil and irrigation water are known to affect

any physiological and metabolic processes, leading to

cell growth reduction [8]. The effect of excess salinity

ar e y ecrease t e growt an transp rat on rates

20 , p otosynt es s an p gment contents 24,25 . H g

exogenous salt concentrations affect seed germination,

water deficit, cause ion imbalance of the cellular ions

esulting in ion toxicity and osmotic stress [14, 15].

e e ect o sa n ty stress on p otosynt et c rate an

water use e c ency was c ose y re ate to ea anatom ca

features. The reduction of mesophyl conductance was

associated with leaf thickness and smaller intercellular

spaces in the mesophyl of salt-stressed leaves, whichay ave ma e t e part towar s t e s tes o CO xat on

ore cu t 23 .

Osmotic effects are due to salt-induced decrease in the

soil water potential [13]. Salt stress induces cellular

accumulation of damaging active oxygen species (ROS).

ROS are g y react ve an tox c to p ants an can ea

o ce eat y caus ng amage to prote ns, mem rane

ipids [15], and nucleic acids [21].

Saline environments are generally correlated with

c anges n p ant p meta o sm 8 . L p perox at on

as een assoc ate w t amages provo e y a var ety

o env ronmenta s tresses. L p perox at on, n uce yfree radicals, is also important in membrane deterioration

8, 11, 15, 17 .

arge num er o p ants spec es accumu ate pro ne

(Pro) in response to salinity stress and that accumulation

may p ay a ro e n com at ng sa n ty stress 2, 18,

19 . However, ata o not a ways n cate a pos t ve

corre at on etween t e osmo yte accumu at on an

adaptation to stress [16].

In the present work, the effects of salt treatment on lipidperox at on, water potent a an water content o ean

were stu e . Lea gas-exc ange an ea anatomy were

a so ana yze .

MATERIAL AND METHODS

Bean see s P aseo us vu gar s L., , cu t var G na,

were sur ace-ster ze w t a 0.5% NaOCL so um

ypoc or te so ut on or 1 m n an t en was e

thoroughly in sterilized water. The seeds were germinated

at dark at 26 C in vermiculite media. After that, they

ere transferred in pots filled by half-strength Hoagland

utr ent so ut on an grown n a growt c am er un er

contro e env ronmenta con t ons. T e con t ons,

aintained during the experiments, were the following:

ight duration 14 hours, light intensity (PAR) 250 mol

s- temperature 22 2 and relative air humidity

60 5%. At t e appearance o t e rst tr o ate ea , an

exper menta es gn w t t ree treatments was arrange

orevery cu t var:

control plants, supplied by of Hoagland solution;

p ants, supp e y o Hoag an so ut on enr c e w t

100 mM NaC ;

plants, supplied by of Hoagland solution enriched with

100 mM Na SO ;

e treatment o p ants w t sa ts cont nue or 7 ays.

L p perox at on was measure as t e amount

Table 1. Effect of salinity on the leaf gas-exchange parameters in bean plants.

Young bean plants are subjected to salt stress by adding 100 mM of NaCl and Na2SO4 to the Hoagland nutrient

solution. PN Net photosynthetic rate [mol (CO2)m-2 s-1]; E Transpiration rate [mmol (H2O) m

-2s

-1]; gs

stomata conductance [mol m-2s-1]; PN/E ratio water use efficiency ; and LA leaf aria (cm2); values are mean

SE. *


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SALINITY AND ITS EFFECTS ON THE PHYSIOLOGICAL RESPONSE OF BEAN (PHASEOLUS VULGARIS L.)

751J. Cent. Eur. Agric. (2008) 9:4, 749-756

o t o ar tur c ac react ve su stance TBARS

eterm ne y t e t o ar tur c ac TBA react on

as described by Heath and Packer (1968) [10]. The leaf

tissues were homogenized in 0.1% (w/v) tricholoacetic

ac TCA . T e omogenate was centr uge at 10,000

g or 20 m n. To m o t e resu t ng supernatant, 4 m o

TCA 20% conta n ng 0.5 w v o TBA were a e . T e

m xture was eate at 95 0C or 30 m n an t en coo en ce o owe y centr ugat on at 10,000 g or 15 m n.

The absorbance was measured at 532 nm and corrected

for 600 nm. The concentration of TBARS was calculated

using an extinction coefficient of 155 mM

-1

cm

-1

.For proline estimation the primary leaves (1 g) were

omogen ze w t 3% aqueous su osa cy c ac an

centr uge at 3,000 g or 10 m n. Pro ne rom t e

supernatant was eterm ne ca or metr ca y w t to uo ,

accordance the method of Bates et al. (1973) [3].

The leaf gas exchange elements were determined

y means o a porta e n rare gas ana yzer LCA-4

Ana yt ca Deve opment Company Lt ., H es on,

England) equipped with a PLCB-4 chamber.

Relative water content (RWC) was calculated according

to Morgan 1986 22 . W o e ea or ea s s 1.5 cm

were we g e mme ate y a ter co ect on or punc ngres we g t, FM an p ace n a Petr s conta n ng

wet filter paper and kept at 4oC in the dark. After 24 h,

the turgor weight (TW) was obtained. For the dry weight

(DM), leaf disks were oven-dried for 24 h at 80-90 C and

we g e .

T e water potent a n eaves was measure w t a

pressure chamber (ELE-International, England).

The leaf area (electronic area meter NEO-3, TU-

So a was measure . T e stomata ce s were measure

by means of a bolt eye-lens - micrometer 15x and an

objective 15x, at magnification 600x. The stomata cells

p otograp s were ta en y means o an eye- ens 16x an

an o ect ve 40x, at magn cat on 640x.

Statistical analysis

A ana yses were one n ve rep cat ons. T e s own

resu ts are mean va ues SE. S gn cant erences

between control and each treatment was determined by

the Students t criterion.

RESULTS AND DISCUSSION

T e e ect o two sa t stresses on p otosynt es s s s own

in Table 1. The gdecreased as a result of submitting to

NaCl or Na SO but the degree of inhibition varied

greatly between the salt sources. Na SO decreased it

to a greater extent 62 % ecrease t an NaC 50 %

n t on . A s gn cant n t on n Pn y eaves

su ecte to ot sa t treatments was o serve , troug

there was no striking difference in the magnitude of

inhibition between the salinity source (62% decrease by

NaCl and 66 % by Na SO ).

Accor ng to Tezara 26 , ot water an sa n ty stress

markedly inhibited Pn and g although the effect of

salinity on these variables was milder than water deficit.

T e resu ts n t e same ta e n cate as we t at was

n te to e esser egree n compar son to Pn 58

ecrease y NaC an 65 % y Na SO , an once

more, the Na SO inhibiting effect was more strongly

expressed.

T e water use e c ency P E was mo e as a resu t

o t e reporte c anges n P an E. In t e eaves o

eans, su ecte to 100 mM NaC an Na SO,t e P E

were reduced to 27-32% compared to that in the control

Table 2. Anatomical characteristics of leaves of Bean exposed for 7 days to 100 mM NaCL and Na2SO4;

values are mean SE.

2. 100 mM

NaCL Na2SO4.). SE.

Parameters Control 100 mM NaCL 100 mM Na2SO4Upper epidermis

Number of stomata 507,6 14,37 782,8 16,49 722,8 13,17

Stomata lenght(m) 22,97 0,32 15,87 0,58 20,76 0,74

Stomata width (m) 18,38 0,13 11,77 0,6 15,34 0,48

Lower epidermis

Number of stomata 75,2 4,22 106,8 5,59 182,8 9,25

Damaged 0 145.28.26 29.66.39

Stomata lenght (m) 27,85 0,49 21,43 0,38 24,63 0,53

Stomata width (m) 19,6 0,25 18,2 0,42 17,79 0,43


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F gure 1. T e e ect o sa n ty on stomata requency.

1.

Control 100 mM NaCl 100 mM Na2SO4

Upperepidermis

Lowerep

idermis

Table 3. Effect of salinity on the parameters of water exchange in bean plants.

Young bean plants are subjected to salt stress by adding 100 mM of NaCl and Na2SO4 to the Hoagland nutrientsolution. WD - Water deficit ( %), RWC - Relative water content (%), - Water potential [-MPa] and P

prline content ( g g-1FM). Values are mean SE. *


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753J. Cent. Eur. Agric. (2008) 9:4, 749-756

mM NaCl and Na SOcaused a increase in the stomata

requency at a out 42% re ate to t e contro p ants

F gure 1 . T e ncrease stomata requency at g er

sa n ty agrees w t t e o servat ons n eans 30 . T s

could be explained ith the dwindlingof the leaf cells as

a result of the xeromorphic structure of the salt-treatedplants. The results show that the stomata size (length and

w t n sa t-treate p ants were re uce too.

Some aut ors ave cons ere t e re uct on n ce s ze

under water stress to be drought adaptation mechanism

[23]. According to Culter et al., [6] the reduction in sell

size appears to be a major response of cells to water

e c ency t at may e cause e t er y roug t or sa n ty

stress. Car m et a ., 5 con rme , as we , t e ecrease

of the stomata size in salt-treated plants. Water status is

highly sensitive to salinity and therefore is dominant in

determining the plant responses to stress [26].

Ta e 3 s ows t e c anges n RWC an water potent a nyoung ean p ants un er var ous sa n ty stresses. A ter 7

days of treatment the osmotic potential of leaves decreased

under saline conditions 98 % (100 mM NaCL) and 138

% (100 mM Na SO ). Generally, decreased lowerin the

secon sa t. T e treatment o p ants w t Na SO causes

more cons era e c anges w t respect an to RWC

14 % lower than the non-treated plants.Dehidratation

symptoms were more evident in the 100 mM Na SO

due to the fact that probably this salt has higher sodium

concentrat ons 2 Na w c ncrease ce u ar water

oss. T e ecrease o RWC an ea s to an ncrease o

t e pro ne content 290 % n case o 100 mM Na SO

treatment. The considerable decrease of RWC and

probably is a result of the structural-functional changes,

Figure 2. Effect of salinity on lipid peroxidation [LP-[nmol (MDA) g-1 (FM)] in shoots and roots in bean plants.

1. LP- nmo MDA g-1 FM

.

12,5

14,73

19,8

19,27

29,9

35,65

0 10 20 30 40 50 60

Control

NaCl

Na2SO4

Variants

nmol MDA g-1 FM

LP shoot LP root

ensuring the plants adaptation to the salt-treatment. In

suc cases, a process o osmot c se -a ustment occurs

n t e p ant ce s, recte towar s t e preservat on o t e

water a ance y means o accumu at on o osmot ca y

active solutes [29], and that is a mechanism reported to

operate in plants in response to salt stress [27]. The resultsfrom our study with respect to the considerable praline

ncrease n ean eaves coor nate we w t t e resu ts

comcern ng t e c anges n t e ot er two parameters

- RWC and . According toCarimi et al., [5] proline is

accumulated in the cytoplasm and its organelles in order

to maintain photosynthesis.

Perox at on o mem rane p s s an n cat on

o mem rane amage an ea age un er sa t stress

conditions. Bean plants were affected by both organs by

means of lipid peroxidation, but in the roots the MDA

content was greater. The results in Figure 2 show as well,

t at t e MDA ncrease more s gn cant y n t e seconsa t 100 mM Na SO 155 % n s oots an 184 % n

roots . L p perox at on s o ten use as an n cator

of salt-induced oxidative stress in sells, by the increase

in MDA level [8].

On the basis of the results obtained during the research,

the following conclusion can be drawn:

T e app e oses o ot sa t types cause stress n

t e young ean p ants, w c oun express on n t e

suppress on o p otosynt es s act v ty an cause

changes in stomata status (conductivity, number and size),

which is a result of the disturbed osmotic processes andthe toxic effect of Cl , SO -and Na . The transpiration

an t e ce water potent a n sa t-treate p ants were


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educed and this tendency was more strongly expressed

n t e case o t e su p ate treatment. T e pro ne an

MDA content was ncrease s gn cant y.

ith respect to all indexes, a stronger toxic effect of

Na SO compared to that of NaCl, was registered, a fact

probably due to the higher concentration of Na in theean p ants t ssues.

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