SALT STRESS RESPONSE

IN PLANTS

Dr Ekamber Kariali
School of Life Sciences,
Sambalpur University, Jyotivihar,
Sambalpur, 768019,Orissa.
Email. ekamberk@rediffmail.com
 In the Dead Sea, no living organisms
can survive except

1. Dunaliela salina & Dunaliela bandawil

(unicellular eukaryotic algae)
2. Asteromonas gracilis
(a bacterium)
due to excess saline water.

Here, Na+ ions are accumulated in the

vacuoles initially and then removed out
side by Na+/H+ antiporter system.
 Definition of Salinity
 Salinity is an increase in the concentration of
dissolved inorganic salts in the soil (soil
solution) or water.
 The main cations include K+, Mg2+, Ca2+, and
Na+, and the main anions are NO3−, HCO3−,
SO42−, Cl−, and CO32−.
 Other components in the highly saline soils
and water are SiO2, Al3+, Sr2+, B, Mo, and
Ba2+ .
 These compounds are reported individually
in mg  L−1 (ppm), mmo1 L−1.
 In agricultural and horticultural section,
total salinity is expressed in the electrical
conductivity of water (ECw) or the soil
extract (ECe).
 Water with electrical conductivity (EC W) less
than 0.7 dS m−1 (desisiemens per meter) or a
salt concentration less than 500 mg L−1 is
classified as drinking water, but non-saline
and it is considered as high-quality water for
irrigation.
 Salt stress that affect soil can vary widely in
their properties and classified in to three
major groups.
(a) Saline soils: pH is less than 8.5, ESP
(Exchangeable Na percentage) less than
15%, and EC more than 4 dS m−1.

(b) Saline-sodic soils: pH is about 8.8, ESP is

more than 15%, and the EC is more than
4 dS m−1.

(c) Sodic soils: pH higher than 8.5, ESP is

more than 15%, and EC is less than 4 dS m−1.
(Sodicity is a term used to describe the amount
of Sodium held in a soil)
 BACKGROUND TO NaCl STRESS
 ~20% of the world’s cultivated land and ~50% of all
irrigated land are affected by salinity (Shuji Yokoi et al.
2002)

 High salinity causes ionic disequilibrium and hyper-

osmotic stress

 Affects negatively on the critical biochemical processes

and quality of produce

 Adversely affects plant growth and development

(germination, seedling growth, flowering, fruiting etc.)

 Ion homeostasis (resistant plants) in saline

environments is primarily dependent on membrane
components (transporter proteins)
EFFECTS OF Na+ TOXICITY IN
PLANTS
 Whole-Plant level

 Plant tissue and cellular level

 Water deficit stress (desiccation or osmotic

stress)

 The ion specific stress results in metabolic

toxicity

 Nutritional deficiencies
Problems caused by salinity in plants
Most of the problems caused by salinity stress
in plants are due to increase conc. of NaCl and
at least 3 major problems in higher plants.
(a) Osmotic effect: High conc. of salt outside
leads to dehydration and causes osmotic or
desiccation stress due to Ψ gradient.
(Physiological drought stress of physiologic dryness)
(b)Specific ion effect: Accumulation of SO2,
Na+, or Cl− in the cells causes damage due to
specific ion effects. High concentrations of Na +
can be replaced with Ca2+ in the plasma
membrane of root hairs in cotton.
Similarly, high salinity stress alters
permeability of Plasma membrane and cause
K+ leakage to surrounding solution.
Salinity also causes inactivation of enzymes,
cell death or even the death of the whole plant.
(c) Nutritional imbalance: The concentration
of essential nutrients like zinc, potassium,
iron, and calcium etc. are largely affected by
Sodium ion. Increasing sodium reduces other
cations in the plant and disrupts the cationic
balance of the plant leading to inhibition of
growth.
A combination of all three effects also occurs.
 Effects of salt stress
Reduction in Growth and Development:
 Immediate response is decrease in leaf area
expansion due to reduced turgor & reduction in
dry and fresh weight of leaves, stems and roots.
 Osmotic effect directly inhibits plant growth due
to inhibition in cell division.
 Osmotic stress reduces water absorption leading
to decrease cell turgor and hence reduced
growth.
 Decreased water availability leads to stomatal
closure and decreased stomatal conductance and
thus reduced photosynthesis causing less TDM
and decreased growth.
 Accumulation of toxic ions causes yellowing and ageing
of leaves causing reduction of photosynthesis and thus
reduction in growth.
 During the late stage of stress, salt accumulates at toxic
levels in the older leaves which is due to lack of salt
transfer for long time to vacuole which inhibit
carbohydrate supply to growing cells and leads to
inhibition of growth.
Seed germination:
 Increased osmotic pressure of soil solution which inhibit
absorption and entry of water into seeds.
 Certain salts like CO3-2, NO3-, Cl-, SO4-2 are toxic to
embryo and seedlings & inhibit germination.
 Salinity affects germination by reducing osmotic potential
of growth medium and ion toxicity that inhibit
metabolism of stored materials required for germination.
Plant Morphology:
 At early stage of stress, decline in growth is very
rapid due to osmotic effect whereas the growth
inhibition in the later stage is relatively slower
due to accumulation of toxic ions in the leaves.
 Continuation of salinity for long time causes
death of leaves, reduction in leaf area, decline in
photosynthetic reserves and yield.
 Leaves becomes colourless and dry wt. loss are
important features of salinity.
 Salinity causes suspension of the apical
meristem, reduction in the diameter of the roots
and their vascular tissue.
Effects on chlorophyll and carotenoids:
 Generally leaf chlorophyll and carotenoid contents
decreased in response to salt stress due to their degradation
mostly by Na.
 Older leaves develop chlorosis and fall down.
 In resistant plants, there is increase in chlorophyll and
carotenoid contents.
Effects on photosynthesis:
 Generally PS-II and Fv/Fm (efficiency of PS-II) are very
sensitive to salts, but again species specific as there is no
reduction of these parameters in resistant plants.
 Accumulation of Na+ &Cl- in chloroplast inhibit
photosynthesis.
 Photosynthetic electron transport appears to be insensitive
to salts, but Carbon metabolism or photophosphorylation
may be affected.
 Enzymes for CO2 assimilation are highly sensitive to NaCl.
Nitrogen metabolism:
 Nitrate reductase is very sensitive to NaCl.
 Concentration of Glycinebetaine increased with
increased level of salinity.
 Proline concentration increases many fold in
response to salinity.
Reproductive growth and Yield:
 Flowering is delayed due to limitation of source
size.
 No. of flowers/panicles is significantly reduced.
 Imbalance of nutrients under salt stress hampers
the hormone synthesis leading to reduction in
the quantity and quality of produce.
Oxidative stress:
 Increased salinity stress causes decreased
photosynthetic rate leading to formation of
more ROS.
 However, resistant plants adjust their leaf
morphology, pigment composition and
activities of different biochemical processes
that prevent oxidative damage.
 Salinity stress also maintains a suitable H2O2
Concentration in the cell that are required for
cell sgnaling.
 Salinity stress in Animals
 Aquatic organisms at the mouth of the river
suffer from salt stress

 These organisms get dehydrated due to

higher osmoticum in the external saline
environment.

 They suffer from a secondary stress (water

deficit) called osmotic stress or desiccation
stress (Physiological drought stress).
GENETIC MODEL SYSTEMS TO STUDY
OSMOTIC STRESS IN PLANTS
 Yeast

 Mesembryanthemum crystallinum- Ice plant,

facultative halophyte

 Craterostigma plantagineum-Resurrection
plant

 Arabidopsis thalliana- Glycophyte

 Thellungiella halophila- Halophyte

 Types of salt stress
1. Primary or Natural salt stress: Due to
accumulation of salts over very long time
through natural processes in soil or water.
It is caused by (a) weathering of native
rocks containing soluble salts. (b)
precipitation of ocean salt by rain and wind.
2.Secondary salinity stress caused by man:
Causes are (a) Clearing the land and replacing
perennial crops with annual ones, (b) irrigation
of salt-rich water, (c)lack of adequate drainage.
 Type of Salt Stress
Although there are various types of salts (i.e.
acid or base) which cause problem for the
Plants, two major salt stresses can cause
significant problem in Plants.
1. Calcium salt stress
2. Sodium salt stress.

 The major problem due to salt stress is the

significant reduction in water potential of
the surrounding environment dehydration.
 When salt conc. is low (toxic ions like Co,
Hg) no or in Ψ Ion stress.
 Calcium salt stress
 Although the major source of salt stress is due
to the presence of excess NaCl, but some time
Ca (CaCl2) deposit also cause localized salt
stress problem.
 The soil where there is Ca deposition, called
Calcareous soil.
( ECOLOGICAL CLASSIFICATION)
 Plants resistant to calcareous soil are called
Calcicoles.
 Plants sensitive to calcareous soil are called
Calcifuse plants.
 Calcium-induced stress injury
 The strain or injury produced in the plant is
usually not due to the primary Ca stress.
 The injury is mainly due to secondary stresses
like increase pH, mineral deficiency etc.

 Excess intake of Ca increase pH

difficulty in absorption of essential
mineral nutrients mineral deficiency stress
(Eg. Lime induced chlorosis is due to Fe
deficiency).
 Physiological classification of the
plants under Ca stress
A. Calciophiles or Calciotrophs ( contains
large amount of soluble Ca in their cell
sap)

B. Calciophobes ( contains very less or no

soluble Ca in the cell sap.

Calciophiles Calcicoles
Calciophobes Calcifuse
 Sodium Salt Stress
 Most of the salinity stress in plants and
animals is primarily due to excess NaCl.
 Excess NaCl is found in many natural
ecosystems like desert, sea, estuaries,
coastal areas etc.
 Plants resistant to extreme salinity are
called Halophytes (Atriplex).
 Plants that can’t grow in presence of high
conc. of NaCl are called Glycophytes or
Sweet plants (Bean, Soyabean, Maize etc.).
 Classification of Halophytes
tolerant to NaCl
Halophytes

Extreme Moderate
euhalophytes or oligohalophytes
true halophytes
• Obligate halophytes : Species can not grow
without saline environment ( Eg. Halobacterium,
but no Angiosperm)
• Facultative halophytes : Halophytic plants that
can grow under normal or low conc. of salt or
non saline environment (Salicornia, Limonium).
 Some halophytic plants inhabit estuaries and sea
shore and adapted to inundated salt water
conditions called mangrove vegetation.

 These plants not only faces the problems of water

uptake, but also suffers from hypoxia or anoxia.
 They develop certain peculiar adaptive mechanisms.

1. viviparous germination ( seed germinate when it

is attached to the stalk within fruit).

2. development of characteristic roots (negatively

geotropic) which bend and come above the water
level for the purpose of aeration called
pneumatophores.
 • Mangroves are ecologically important for
coastal erosion.
 Glycophyte plants which can tolerate some
levels of salt are called Salt tolerant non
halophytes.
 Salt stress injury
 Primary direct salt injury:
Generally, there is no direct salt injury,
But, salt shock or osmotic shock can occur, when
salt conc. is extremely high.
 Primary salt injury must be specific toxic effects
of salt either
(a) directly on external P.M. of cell, or,
(b) after penetration through PM into protoplast.
Expt.1
1.Two plants with identical wt.
and identical ages.
2. Pot A treated with pure water
to maintain Ψ =0 and Pot B Ψ=
-10bar by adding NaCl.
3. Plant in Pot 2 will be
dehydrated and injured.
Plants in Pot B will be
dehydrated and injured which
might be due to
(a) Decrease inΨ due to NaCl
or, (b) Direct effect of salt or
salt shock.

Expt. 2:
• Similar experiments conducted in pot A and pot B.
• Pot A will be treated with PEG (organic solute) not absorbed by plants,
but can decrease in Ψ = -10 bar.
• Pot B will be treated with NaCl which lowers the Ψ =-10bar
and is absorbed by the plants.
• Plant in pot B is severely injured and wilted, but in A, plant grow well.
• Hence, Injury of plant in pot B is not due to osmotic potential, but is
due to the direct effect of salt (salt shock).
Different salts requires diff. conc. to produce same injury also.
So, NaCl, Na2SO4, K2SO4 produce more injury than KCl or MgCl2.
Often, Injurious NaCl becomes non-injurious in presence of CaCl2
(10:1) called ‘Ion antagonism’.
Primary indirect salt injury:
(a) inhibition of growth and development
( NaCl inhibited mitotic activity in Crepis capillaris,
growth inhibition in wheat is due to reduced tillering)
(b) metabolic disturbances (photosynthesis, respiration,
protein synthesis etc.)
(c) Alteration in enzyme activities
(d) Production of toxins (putrescine, cadaverine etc.
Products of disturbed N2 metabolism.
In resistant plants, these compounds will be metabolized
to produce Proline.
Secondary stress injury:
(a) Osmotic stress: Cells dehydrated due to higher
osmoticum of salts in the external environment leading
to desiccation or osmotic stress (physiological drought
stress).
(b) Mineral deficiency stress: In presence of excess Na salt,
K may not be absorbed due to similar properties. Hence,
plants suffer from Potassium deficiency.
 Alteration in Enzymatic activities
 Catalse, peroxidase increase or decrease)
 Chlorophyllase at low conc. and at high conc.
 Nitrate reductase,ribonuclease always at high salt
conc.
 Phosphohydrolases insensitive to salt conc.

 The activity of some enzymes increases whereas the

activity decreases for other enzymes which might be due
to two antagonistic effects of salts on proteins.

(a) salts tend to break the electrostatic bonds.

(b) salts tend to increase the hydrophobic interactions.

How does Salts enter into Plants
 MECHANISMS OF CELLULAR RESISTANCE
TO HIGH Na+ IN PLANTS
AVOIDANCE
 Sodium exclusion
(Physiological impermeability of salts towards membrane,
filtration by root cell membrane, passive process of
avoidance, Eg. Red mangrove)

 Sodium Extrusion
(Ion pumps, salt pumps collectively called extrusion
pumps which pumped out the salts from the
cytoplasm by utilisation of ATP. So it is an active
process.
 Extrusion mechanism is some times localised in salt
glands (Mesembryanthemum crystallinum) or salt
bladders (Atriplex) in the leaves or cuticle (Tamarix).
 Dilution: (Absorption of water), Succulence (formation of
vacuolated cells for water storage) etc.
Recently, SOS (salt overly sensitive) genes have
been identified in Arabidopsis mutants (extremely
sensitive to salts) that mediate a novel signaling
pathway in ion homeostasis and salt tolerance.
 The primary P-ATPase uses the energy of
ATP hydrolysis to pump H+ ions out of
the cell generating an electrochemical
proton (H+) gradient.
 The proton gradient operates a secondary
active transport by Na+/H+ antiporter
which removes Na+ to out of the cell
coupled with the movement of H+ into the
cell.
 The primary active V-ATPase and V-
PPase (pyrophosphatase) energize the
tonoplast for secondary active transport
of Na+ into the vacuole by the Na+/H+
antiport.
 High Na+ stress initiates a calcium signal
(increase Ca ion conc.) in side the cell.
 In this pathway, there is a calcium-binding
protein called SOS3 which senses cytosolic
calcium changes.

 SOS3 physically interacts with and

activates a protein kinase, SOS2.

 Now, this SOS3/SOS2 kinase complex

phosphorylates and activates the transport
activity of the plasma membrane Na +/H+
exchanger encoded by the SOS1 gene.
 P-ATPase energises the Na+/H+ antiporter
by ATP hydrolysis creating a H+ gradient
out side the PM that helps Na+ to go out.

 In addition, SOS3-SOS2 may activate or

suppress the activities of other transporters
involved in Na+ homeostasis.
 Avoiding the effects of stresses is important, as it
will reduce the risk of enhanced ROS production.
 Mechanisms that might reduce ROS prodcution
include
(1)anatomical adaptation such as leaf curling, hiding
of stomata.
(2) physiological adaptations such as C4/CAM
metabolism.
(3)Alternative channeling of electrons in electron
transport chains by alternative oxidases.
 Plants regulate their internal ROS by two different
mechanisms (a) scavenging or (b) elimination.
 Most of the transgenic improvements in salt
tolerance reported have been achieved through the
detoxification strategy.
 Tolerance
 Excess salts leads to dehydration or water deficit
stress and thus tolerance can be
(a) dehydration avoidance ( osmoregulation)
i. Active uptake of salt ions
ii. Synthesis of organic solutes
(b) dehydration tolerance
 Intracellular compartmentation to vacuoles
Organ level- high salts in roots compared to shoots
especially leaves; cellular level- high salts in
vacuoles than cytoplasm & thus protects
enzymes.
 Tolerance by metabolising the toxins or ROS and
produce proline, glycinebetaine etc.
 Metabolic shift (C3 C4 or C3 CAM as salt ions
like Cl- activates PEP carboxylase)
 If there is damage response, repair of injuries.
 Salt stress can produce
secondary Oxidative stress
 Salt stress can produce large amount of ROS.
 Chloroplasts, mitochondria, peroxisomes are
the primary sites of ROS production.
 ROS can cause damage to macromolecules,
but also acts as signal for the activation of
stress-response and defense pathways.
 Over-accumulation however results in cell
death either by oxidative processes or
activating programmed cell death (PCD).