Mineral
nutri+on


&
uptake

 Soils
and
plant
nutri+on

•  14
essen+al
mineral
nutrients

–  N,P,K,S,Ca,Mg,Fe,Cl,Mn,B,Zn,Cu,Mo,Ni

•  3
essen+al
non‐mineral
nutrients

–  C,H,O


–  What
makes
these
nutrients
essen+al?

Magnesium
and
photosynthesis


•  Chlorophyll
(Chl)
is
a
tetrapyrrole
macrocycle
containing

Mg2+,
a
phytol
chain,
and
a
characteris+c
fiMh
ring

•  The
methyl
group
at
the
C7
posi+on
of
Chl
a
is
replaced
by
a

formyl
group
in
Chl
b.

Nitrogen
and
amino
acids

Nitrogen,
phosphorus
&
DNA

Potassium
and
cell
func+on

Macronutrients
vs.
micronutrients

Mineral
nutrients
exist
as
ions

Mineral
nutrient
transport

Soil
pH
and
mineral
nutri+on


Different types of plants have different soil pH requirements

 Macronutrients

Element Symbol Source
•  Macronutrients
are

needed
in
rela+vely
 Oxygen O Air/Water
large
amounts
by
 Hydrogen H Air/Water
plants.

 Carbon C Air/Water
Nitrogen N Soil
Phosphorus P Soil
Sulfur S Soil
Potassium K Soil
Calcium Ca Soil
Magnesium Mg Soil
 Micronutrients

•  Macronutrients
are
 Element Symbol Source
needed
in
rela+vely

Iron Fe Soil
small
amounts
by

plants.

 Manganese Mn Soil
•  They
are
usually
 Boron B Soil
supplied
by
 Molybdenum Mo Soil
fer+lizers.
 Copper Cu Soil
Zinc Zn Soil
Chlorine Cl Soil
 Nutrient
Deficiency
Symptoms


•  Nutrient
deficiency
symptoms
usually
appear

on
the
plant
when
one
or
more
nutrients
are

in
short
supply.


•  In
many
cases,
deficiency
may
occur
because

an
added
nutrient
is
not
in
the
form
the
plant

can
use.


•  Deficiency
symptoms
for
specific
elements
are

included
on
the
"Key
to
Nutrient
Disorders".


 Deficiency
Symptoms
‐
N

•  General
chlorosis.


•  Chlorosis
progresses

from
light
green
to

yellow.


•  En+re
plant
becomes

yellow
under
prolonged

stress.


•  Growth
is
immediately

restricted
and
plants

soon
become
spindly

and
drop
older
leaves.

 h\p://plantsci.sdstate.edu/woodardh/soilfert/
Nutrient_Deficiency_Pages/soy_def/SOY‐N1.JPG

 Deficiency
Symptoms
‐
P

•  Leaves
appear
dull,
dark

green,
blue
green,
or
red‐
purple,
especially
on
the

underside,
and
especially

at
the
midrib
and
vein.

•  Pe+oles
may
also
exhibit
 h\p://www‐unix.oit.umass.edu/~psoil120/images/tomatox2.jpg


purpling.
Re‐stric+on
in

growth
may
be
no+ced.


h\p://www.ext.vt.edu/news/periodicals/
vi+culture/04octobernovember/photo3.jpg

 Deficiency
Symptoms
‐
K


•  Leaf
margins
tanned,

scorched,
or
have
necro+c

spots
(may
be
small
black

spots
which
later
coalesce).


•  Margins
become
brown
and

cup
downward.

•  
Growth
is
restricted

•  
Mild
symptoms
appear
first

on
recently
matured
leaves.


h\p://www.ipm.iastate.edu/ipm/icm/files/images/antonio004f.jpg

 Deficiency
Symptoms
‐
Ca

•  Growing
points
usually

damaged
or
dead
(die

back).


•  Margins
of
leaves

developing
from
the

growing
point
are
first
to

turn
brown.



h\p://hubcap.clemson.edu/~blpprt/acid_photos/
BlossomEndRot.JPG

 Deficiency
Symptoms
‐
Mg

•  Marginal
chlorosis
or

chloro+c
blotches
which
later

merge.


•  Leaves
show
yellow
chloro+c

interveinal
+ssue
on
some

species,
reddish
purple

progressing
to
necrosis
on

others.


•  Younger
leaves
affected
with

con+nued
stress.


•  Chloro+c
areas
may
become

necro+c

•  Symptoms
usually
occur
late

in
the
growing
season.


h\p://quorumsensing.ifas.ufl.edu/HCS200/images/
deficiencies/‐Mgcq.jpg

 Deficiency
Symptoms
‐
S

•  Leaves
uniformly
light
green,
followed
by
yellowing

and
poor
spindly
growth.


•  Uniform
chlorosis
does
not
occur



h\p://www.ces.ncsu.edu/plymouth/cropsci/ h\p://www.ag.ndsu.nodak.edu/aginfo/
graphics/sulfur2.jpg
 entomology/ndsucpr/Years/2007/june/7/soils.jpg

 Deficiency
Symptoms
‐
Cu


•  Leaves
wilt,
become

chloro+c,
then
necro+c.


h\p://images.google.com/url?q=h\p://ipm.ncsu.edu/
Scou+ng_Small_Grains/Grain_images/
fig4.jpg&usg=AFQjCNE2vzRwrqp65VR_xKRlo2LQOgWI3g

 Deficiency
Symptoms
‐
Fe


•  Dis+nct
yellow
or
white

areas
appear
between

veins,
and
veins

eventually
become

chloro+c.


•  Symptoms
are
rare
on

mature
leaves.



h\p://bexar‐tx.tamu.edu/HomeHort/F1Column/
2003Ar+cles/Graphics/iron%20chlorosis.jpg

 Deficiency
Symptoms
‐
Mn


•  Chlorosis
is
less
marked

near
veins.


•  Chloro+c
areas

eventually
become

brown,
transparent,
or

necro+c.

•  Symptoms
may
appear

later
on
older
leaves.

h\p://www.ca.uky.edu/HLA/Dunwell/KHC/110‐122.JPG

 Deficiency
Symptoms
‐
Zn

•  Leaves
may
be
abnormally
small
and
necro+c.


•  Internodes
are
shortened.


h\p://agri.atu.edu/people/Hodgson/FieldCrops/
Mirror/Nutrient%20Def_files/slide24.jpg


h\p://plantsci.sdstate.edu/woodardh/soilfert/
Nutrient_Deficiency_Pages/corn_def/CORN‐
ZN1.JPG

 Deficiency
Symptoms
‐
B

•  Young,
expanding
leaves

may
be
necro+c
followed

by
death
of
growing

points.

•  Internodes
may
be
short,

especially
at
shoot

terminals.



h\p://www.canr.msu.edu/vanburen/ffc12.jpg

 Plant Nutrition :
Membrane energetics and
transport, potassium nutrition and
sodium toxicity
 Plant
nutri+on:
Introduc+on

Plants
are
~70
to

>90%
water
by

weight

 N


Nitrogen


P
Phosphorus

CO2,

 K
Potassium

photo‐synthesis
 Ca
Calcium

Mg
Magnesium

S
Sulfur

42%
Carbon
 7%
Other,

Si
Silicon

from
soil
 Cl
Chlorine

Other

7%
Hydrogen

These
elements
are
obtained

44%
Oxygen
 mainly
from
soil,
are
oHen

93%
of
plant
 referred
to
as
mineral

dry
mass
is

composed
of
C,

nutrients,
and
are
the
subject

H2O
water

O
and
H
 of
the
topic
Plant
NutriJon

 Plants assimilate mineral nutrients
from their surroundings
Nutrient assimilation is very
energetically demanding –
the nutrients have to be
moved against a
concentration gradient and
often a charge gradient

Tracheophytes

K+

Algae
 NO3‐

NO3‐

NO3‐


K+
 K+


PO43‐
 Bryophytes
 K+


K+

K+
 PO43‐

PO43‐
 PO43‐
 PO43‐

PO43‐

NO3‐


K+
 K+

 Nutrient uptake, assimilation and
utilization involve many processes
Nutrient
 Nutrient
usage

acquisiJon
 efficiency

efficiency

Root

X
 R‐X

exudates

Root
system

AssimilaJon
and

architecture

Intercellular
 remobilizaJon

transport
 efficiency


P
efficiency

Transporters

P
and
pumps

Regulatory
and

Symbioses
 NH3
 homeostaJc

networks

N N

Rhizosphere

microbiota

 Nutrients removed from soils can
be replenished with fertilizers
Plants
remove
nutrients
from
the
soil

FerJlizers
can
be

1000
 Total
nutrient
requirement

 complex

waste

products
or
refined


Cotton
800
 blends
of
nutrient

Wheat

salts


Rice
Corn


600
 Sulfur

Soy

Kg/ha


Magnesium

400

Potash


200
 Phosphate
 Most
fer6lizers


0
 Nitrogen
 contain
nitrogen
(N),

400
 Typical
ferJlizer
applicaJon
 phosphorus
(P)
and

potassium
(K).
Some

include
other

Kg/ha


200
 elements


0


Source: USGS
 Global mineral nutrient resources
are unevenly distributed

Supply
>
Demand

Supply
<
Demand


N

P2O5

K2O


FAO (2011) Current world fertilizer trends and outlook to 2015.

 The global trade in fertilizers is worth
billions of dollars annually

Ammonium
 Urea
 Potash
 Diammonium
 Monoammonium
 Phosphate
rock
 Sulfur
 Sulfuric


phosphate
 phosphate
 acid

IFIA
 How much is the right amount of
fertilizer to apply to a field?
CulJvaJon

Species
/
variety
of
 pracJces:
Is

plant:
Different
 unharvested

plants
have
 material
removed,

different
needs
 or
leM
to
replenish

the
soil?



AbioJc
and
bioJc

Soil
characterisJcs:

 factors:
Temperature,

Residual
nutrients,
 rain,
stress
and
pests
or

rate
of
nutrient
 pathogens
affect

leaching,
pH,
par+cle
 nutrient
needs

size,
presence
of

microbes
etc.
affect
 Developmental
stage
affects
plant
needs

op+mal
applica+on

Financial
consideraJons:
Balancing

the
cost
of
fer+lizers
with
the
gain

reaped
from
their
use

Photo by Michael Russelle.
 Fertilizer use can cause
environmental and health problems
Nitrogen
fixa6on
is
energy
 Human
and
animal
waste
can

demanding
 spread
disease
 N
 N
 O


Nitrous
oxide
(N2O)
derived

from
fer6lizer
is
a
major

Transport
requires
energy
 greenhouse
gas


Plants need
nutrients, but their
application isn’t
Phosphate
and
potash

always optimal or
mining
is
destruc6ve

sustainable – how
can plant science
contribute to
Nutrient
runoff
pollutes
 better practices?
waterways
and
can
lead
to

eutrophica6on
 Image source: Lamiot; Alexandra Pugachevsky
 Nutrient
uptake
and
transport:
Overview


Nutrients must be Membrane transport can

Cross
membrane
 consume 1/3 of a cell’s
transported across
into
living
cell
in
 metabolic energy (or more)
membranes to enter root
hair
 Symplas6c
or

the plant
transcellular
pathway
 Casparian
strip


Apoplas6c
pathway
 Bidirec6onal
transport
between

Pumps,
channels
and

Cross
membrane
 xylem
parenchyma
cells
and

carriers
are
the

apoplas6c
transpira6on
stream

molecular
mediators
 into
living
cell
in
at

of
these
processes
 endodermis

 Plants assimilate mineral nutrients
mainly as cations or anions
MACRONUTRIENTS
 MICRONUTRIENTS

μmol
/
g

 Element
 Assimilated

 μmol
/
g

 Element
 Assimilated


(dry
wt)
 form
 (dry
wt)
 form

250
 Potassium
(K)
 K+
 2
 Iron
(Fe)

 Fe3+,
Fe2+

1000
 Nitrogen
(N)
 NO3‐,
NH4+
 0.002
 Nickel
(Ni)

 Ni2+

60
 Phosphorus
 HPO42‐,
 1
 Manganese
 Mn2+

(P)
 H2PO4‐
 (Mn)


30
 Sulfur
(S)
 SO42‐
 0.1
 Copper
(Cu)

 Cu2+

80
 Magnesium
 Mg2+
 0.001
 Molybdenum
 MoO42+

(Mg)

 (Mo)

125
 Calcium
(Ca)

 Ca2+
 2
 Boron
(B)
 H3BO3

3
 Chlorine
(Cl)

 Cl‐

0.3
 Zinc
(Zn)
 Zn2+

See Taiz, L. and Zeiger, E. (2010) Plant Physiology. Sinauer Associates; Marschner, P. (2012) Mineral Nutrition of Higher Plants. Academic Press, London
 Nutrients are concentrated in the
plant relative to the environment
Energy
is
expended
to
assimilate
nutrients
against
a
 The
driving
force
of
the

steep
concentra+on
gradient
 nutrient’s
chemical

gradient
is
outwards


[H2PO4‐]o
 [NO3‐]o
 [NH4+]o


Soil
abundance
(ranges
 [K+]o
 [HPO42‐]o
 <100
μM
–

 <100
μM
–


or
typical
values)
 0.1
–
1
mM
 <
1
μM
 >1
mM
 >1
mM


Cell

[K+]i
 [H2PO4‐]i
 [NO3‐]i
 [NH4+]i

50
‐
100
mM
 [HPO42‐]i
 10
mM
 ~1
mM

5
‐
10
mM

 Transport can be down or against an
electrochemical gradient
Down
an
electrochemical
gradient

 Against
an
electrochemical
gradient

(Diffusion
or
facilitated
diffusion)
 (Ac+ve
transport)

Symport
 AnIport


OUT


IN


ATP
 ADP
+
Pi

Through
 Through
 Through
carrier

Secondary
ac6ve

membrane
 channel

Primary
ac6ve
transport:
 transport:
Indirectly

Directly
coupled
to
ATP
 coupled
to
ATP
hydrolysis

hydrolysis

 Solutes cross membranes through different
types of transporters

Pumps: ATP
 ADP


•  Move solutes against a
ATP
 chemical or charge gradient
X
 •  Couple transport to hydrolysis
of ATP or pyrophosphate
ADP
 X

Pumps
are
oMen
drawn
like

lollipops,
with
a
large

cytoplasmic
cataly+c

domain


The
mul+subunit
vacuolar

proton
pump
VH+‐ATPase


Reprinted from Schumacher, K. and Krebs, M. (2010). The V-ATPase: small cargo, large effects. Curr. Opin. Plant Biol. 13: 724-730 with permission from Elsevier.
 Solutes cross membranes through
different types of transporters
Channels:
•  are protein-formed holes in the membrane X

•  can be open or closed
X
 • 
• 
move one type of solute at a time
do not provide an energy source for the X

Channels
are
oMen
drawn

movement; solutes can only move down as
two
adjacent
ovals
(or
a

their electrochemical gradient cross‐sec+on
of
a

doughnut)


Reprinted from Long, S.B., Campbell, E.B. and MacKinnon, R. (2005). Crystal structure of a mammalian voltage-dependent Shaker family K+ channel. Science. 309: 897-903 with permission from AAAS.
 Solutes cross membranes through
different types of transporters
Carriers
/


Coupled
Transporters

H+
 • are
membrane
proteins

X
 H+
 H+

• can
be
ac+ve
or
inac+ve

X
 • can
move
more
than
one
solute
at
a
+me

X

• The
driver
(usually
H+
in
plants)
moves
down
its

Coupled
transporters
are

electrochemical
gradient,
which
provides
the
energy
 oMen
drawn
as
circles

for
the
co‐transported
solute’s
transport
 with
arrows
indica+ng
the

direc+on
of
flow
for
each

ion


Schema+c
domain
structure
(L)

and
Top‐down
(R)
views
of
an

HKT1
Na+
transporter


Cotsaftis, O., Plett, D., Shirley, N., Tester, M. and Hrmova, M. (2012). A two-staged model of Na+ exclusion in rice explained by 3D modeling of HKT transporters and alternative splicing. PLoS ONE. 7: e39865. Chérel,
I., Lefoulon, C., Boeglin, M. and Sentenac, H. (2014). Molecular mechanisms involved in plant adaptation to low K+ availability. J. Exp. Bot. 65: 833-848, by permission of Oxford University Press.
 Pumps, channels and carriers are
also involved in nutrient distribution
•  Nutrient
uptake
is

just
the
first
step


•  The
assimilated

nutrients
have
to
be

transported
to
where

they
are
needed,

including
leaves
and

seeds


•  The
vacuole
is
an

important
storage

compartment


Reprinted from Ahmad, I. and Maathuis, F.J.M. (2014). Cellular and tissue distribution of potassium: Physiological relevance, mechanisms and regulation. J. Plant Physiol. 171: 708–714 with permission from Elsevier.
Nutrient transport requires energy and selective
transporters

Proton
(H+)
gradient


Plant cells pump protons

out to make proton and
charge gradients

‐
 ‐
 ‐

+
+
+

Charge
gradient


The electrical (charge) and

K+
 proton gradients drive the
movements of ions across
H+

membranes through selective
K+
 transporters
 Plant and fungal PM H -ATPases
+

are members of a larger family

H+

3Na+

 2K+



ATP
 ADP
+
Pi


3Na+


2K+



Plant
and
fungal

plasma

Mammalian

 membrane


Na+/
K+‐ATPase
 H+‐ATPases


Reprinted by permission from Macmillan Publishers Ltd from Kühlbrandt, W. (2004). Biology, structure and mechanism of P-type ATPases. Nat. Rev. Mol. Cell Biol. 5: 282-295; see also Baxter, I., Tchieu, J., Sussman,
M.R., Boutry, M., Palmgren, M.G., Gribskov, M., Harper, J.F. and Axelsen, K.B. (2003). Genomic Comparison of P-Type ATPase Ion Pumps in Arabidopsis and Rice. Plant Physiol. 132: 618-628.
 Several differentially expressed genes
encode plant PM H+-ATPases
Phylogeny
showing
func+onal
 AHA3
is
highly
expressed
in
phloem
companion
cells

genes
in
Arabidopsis

(AHA)
and

four
in
the
liverwort
Marcan6a
 Nega6ve

polymorpha
(MpHA)
 control


An6body
stain


Other
PM
H+‐ATPase‐encoding
genes
are
expressed

O
 in
other
+ssues,
and
many
are
upregulated
by
stress

or
other
factors

Arango, M., Gévaudant, F., Oufattole, M. and Boutry, M. (2003). The plasma membrane proton pump ATPase: the significance of gene subfamilies. Planta. 216: 355-365.Okumura, M., Inoue, S.-i.,
Takahashi, K., Ishizaki, K., Kohchi, T. and Kinoshita, T. (2012). Characterization of the plasma membrane H+-ATPase in the liverwort Marchantia polymorpha. Plant Physiol. 159: 826-834. DeWitt, N.D.
and Sussman, M.R. (1995). Immunocytological localization of an epitope-tagged plasma membrane proton pump (H+-ATPase) in phloem companion cells. Plant Cell. 7: 2053-2067. See also Okumura, M.,
Takahashi, K., Inoue, S.-i. and Kinoshita, T. (2012). Evolutionary appearance of the plasma membrane H+-ATPase containing a penultimate threonine in the bryophyte. Plant Signal. Behav. 7: 979 - 982.
Plant PM H+-ATPases are essential for
nutrient uptake and allocation
Through their combined
actions, PM H+-ATPases
contribute to the movement of
nutrients throughout the plant

Sondergaard, T.E., Schulz, A. and Palmgren, M.G. (2004). Energization of transport processes in plants. Roles of the plasma membrane H+-ATPase. Plant Physiol. 136: 2475-2482.
 PM H -ATPases
+ have diverse
physiological roles

•  Loss of function of the PM-H+-ATPase is lethal

•  PM H+-ATPases are required for
•  nutrient assimilation and transport,
•  cell growth, turgor and movement
•  guard cell dynamics
Sondergaard, T.E., Schulz, A. and Palmgren, M.G. (2004). Energization of transport processes in plants. Roles of the plasma membrane H+-ATPase. Plant Physiol. 136: 2475-2482.
 Vacuolar
pumps
pump
protons
into

the
vacuole
and
endocompartments

H+

pH
5
‐
6


pH
7.5

H+

ATP
 ADP
+
Pi

H+

PPi
 2
x
Pi


Protons
are
pumped
into
the
vacuole

by:

• 
Vacuolar
H+‐ATPases
(VH+‐ATPases)

and

 H+

• Vacuolar
pyrophosphatases
(H+‐
H+

pH
3
‐
6
 Em
=
~
‐30
mV

PPases)


Sze, H., Li, X. and Palmgren, M.G. (1999). Energization of plant cell membranes by H+-pumping ATPases: Regulation and biosynthesis. Plant Cell. 11: 677-689.
Isayenkov, S., Isner, J.C. and Maathuis, F.J.M. (2010). Vacuolar ion channels: Roles in plant nutrition and signalling. FEBS letters. 584: 1982-1988.
 VH+-ATPases contribute to growth, salt
tolerance & ion uptake / storage
Phenotypes associated with
decreased VH+-ATPase activity:
•  Decreased growth rate
•  Male sterility
•  Altered nutrient storage
capabilities

A
mutant
lacking
func+onal
tonoplast


VH+‐ATPase
is
sensi+ve
to
Zn2+
toxicity

(cannot
sequester
it
into
vacuole)


Decreased
growth
rate
in
carrot
plants

expressing
an
an+sense
VH+‐ATPase
A

construct


Gogarten, J.P., Fichmann, J., Braun, Y., Morgan, L., Styles, P., Taiz, S.L., DeLapp, K. and Taiz, L. (1992). The use of antisense mRNA to inhibit the tonoplast H+ ATPase in
carrot. Plant Cell. 4: 851-864. Krebs, M., Beyhl, D., Görlich, E., Al-Rasheid, K.A.S., Marten, I., Stierhof, Y.-D., Hedrich, R. and Schumacher, K. (2010). Arabidopsis V-
ATPase activity at the tonoplast is required for efficient nutrient storage but not for sodium accumulation. Proc. Natl. Acad. Sci. USA. 107: 3251-3256.
 The VH+-ATPases have different roles in
different compartments
In
Arabidopsis,
three
genes

encode
VHA‐a,
and
their
 Mutants
lacking
one
or
the
other

gene
products
localize
to
 isoform
reveal
that
the


different
subcellular
 VH+‐ATPases
have
different
roles
in

compartments
 different
compartments


VHA‐a2
localizes
to
the
 VHA‐a1
localizes
to
the
trans‐ TEM
showing
VHA‐a1(black


vacuolar
membrane
 Golgi
network
 dots)
localized
in
TGN

Nishi, T. and Forgac, M. (2002). The vacuolar (H+)-ATPases — nature's most versatile proton pumps. Nat. Rev. Mol. Cell Biol. 3: 94-103. Dettmer, J., Hong-Hermesdorf,
A., Stierhof, Y.-D. and Schumacher, K. (2006). Vacuolar H+-ATPase activity is required for endocytic and secretory trafficking in Arabidopsis. Plant Cell. 18: 715-730.
VH+-ATPases have multiple functions in
plants and animals
VH+-ATPases have roles in
pH homeostasis in
endomembranes and also
in membrane trafficking

Functions in plants
•  Development
•  Cell expansion
•  Nutrient assimilation

Functions in humans
•  Development
•  Kidney function
•  Bone resorption
•  Tumor cell metastasis

Reprinted from Schumacher, K. and Krebs, M. (2010). The V-ATPase: small cargo, large effects. Curr. Opin. Plant Biol. 13: 724-730 with permission from Elsevier.
 Plants have 2 types of H+-PPases

Type
2:


Golgi‐localized

K+
insensi+ve

Strongly
inhibited
by
Ca2+


Blue indicates
eubacteria
Red indicates
Archaea
Green indicates
eukaryotes Type
1:


Tonoplast‐localized

Eukaryotes with H+- Require
K+
for
ac+vity

PPases are limited to Moderately
inhibited
by
Ca2+

plants and green
algae (circled in
green) and parasitic
protists (circled in
orange)

Reprinted from Drozdowicz, Y.M. and Rea, P.A. (2001). Vacuolar H+ pyrophosphatases: from the evolutionary backwaters into the mainstream. Trends Plant Sci. 6: 206-211 with permission from Elsevier; Gaxiola, R.A.,
Sanchez, C.A., Paez-Valencia, J., Ayre, B.G. and Elser, J.J. (2012). Genetic manipulation of a “vacuolar” H+-PPase: From salt tolerance to yield enhancement under phosphorus-deficient soils. Plant Physiol. 159: 3-11.
 H+-PPases have many physiological
roles
Altering H+-PPase expression affects:
Salinity and drought tolerance
Nutrient uptake
Auxin transport
Phosphate uptake
Fruit ripening …….

Plants
overexpressing
H+‐PPase
show
 Accelerated
fruit
ripening
in
tomato
plants

enhanced
drought
tolerance
 overexpressing
H+‐PPase


Gaxiola, R.A., Li, J., Undurraga, S., Dang, L.M., Allen, G.J., Alper, S.L. and Fink, G.R. (2001). Drought- and salt-tolerant plants result from overexpression of the AVP1 H+-pump. Proc. Natl, Acad. Sci. USA. 98:
11444-11449. Yang, H., Zhang, X., Gaxiola, R.A., Xu, G., Peer, W.A. and Murphy, A.S. (2014). Over-expression of the Arabidopsis proton-pyrophosphatase AVP1 enhances transplant survival, root mass, and fruit
development under limiting phosphorus conditions. J. Exp. Bot. 65: 3045-3053 by permission of Oxford University Press.
K+ and Na +- “The twins”. So alike
yet so different
NaCl
toxicity

Potassium
deficiency


Sodium (Na) and potassium (K):

•  Same column of the periodic table
•  Both have a single electron in the
outer shell so form monovalent
cations
•  Both are very abundant elements

K
And
yet,
potassium
is
an

essen+al
nutrient,
and
sodium

frequently
is
toxic


Benito, B., Haro, R., Amtmann, A., Cuin, T.A. and Dreyer, I. (2014).The twins K+ and Na+ in plants. J. Plant Physiol. 171: 723–731. FAO
 Potassium
uptake,
transport
and

homeostasis

Potassium
is
an
essenJal
macronutrient

Enhances
fertility Maintains turgor
and reduces wilting
Promotes stress
tolerance Regulates
stomatal
Regulates Symptoms
of
potassium

conductance,
enzyme activities deficiency

photosynthesis
and transpiration
Strengthens
cell walls
Maintains ionic
Stimulates homeostasis
photosynthate
[K+] in soil = ~0.1 – 1 mM
translocation
[K+] in plant cell
cytoplasm = ~100 mM

See Wang, M., Zheng, Q., Shen, Q. and Guo, S. (2013). The critical role of potassium in plant stress response. Intl. J. Mol. Sci. 14: 7370-7390; Sin Chee Tham /Photo; Purdue extension; Onsemeliot.
 Potash provides for fertilizers, K +

which supplement natural sources

Water
pumped

underground

Water
with

dissolved
K+
 Potash

salts
returned
to
 fer6lizer

surface
 applica6on


0.1 – 0.2% soil manure


Salts
recovered


solution K+ decomposi6on

by
evapora6on

1
–
3%

exchangeable
 Terrestrial

salts
 cycle:
Plant
/

Animal
/
Soil

Underground
reserves

90
–
98%

insoluble

minerals


Adapted from International Potash Institute
 Potash prices can be volatile and
Canada
is
#1
in

there are few suppliers
produc+on
 Russia
is
#2
in

(11.2
Mt)
and
 UK Belarus
5.5 Mt produc+on

reserves
(4,400
 0.4 Mt
750 Mt (7.4
Mt)
and

Mt)
 22 Mt
reserves
(3,300

Mt)

Spain 1.0
0.4 Mt 6

US 20 Mt cm

China
1.1 Mt
Germany 3.2 Mt
130 Mt
3.3 Mt 210 Mt World
150 Mt reserves
9500 Mt
Jordan
Brazil Israel 1.4 Mt
Chile 3.2 Mt 2.0 Mt World
40 Mt
0.8 Mt 210 Mt 40 Mt production
130 Mt
(2011)
37 Mt

Adapted from International Potash Institute

 Potassium
is
an
essen+al
plant

nutrient

K+
is
a
counter
ion
for
nega+vely

charged
molecules
including
DNA

and
proteins


K+
moves
in
and
out

of
the
vacuole

through
specific

transporters


As
the
major
ca+on
in

the
vacuole,
K+

contributes
to
cell

expansion
and

movement,
including

K+
uptake
 K+
is
a
cofactor
for
 that
of
guard
cells


involves
high
 some
enzymes

and
low
affinity

transporters

Reprinted from Maathuis, F.J.M. (2009). Physiological functions of mineral macronutrients. Curr. Opin. Plant Biol. 12: 250-258 with permission from Elsevier.
 There are several types of coupled
transporters for K+
KT/KUP/HAK
transporters
are
responsible
 Some
members
of
the
large
CPA
(Ca+on
Proton

for
much
of
the
high‐affinity
uptake
into
 An+porter)
family
contribute
to
K+
uptake

roots.
There
are
13
genes
in
Arabidopsis

Blue
indicates

and
27
in
rice
 preferen+al

K+

uptake



They
are
K+
/
H+
 K+
 H+


symporters

K+
/
H+
an+porters

Chérel, I., Lefoulon, C., Boeglin, M. and Sentenac, H. (2014). Molecular mechanisms involved in plant adaptation to low K+ availability. J. Exp. Bot. 65: 833-848, by permission of Oxford
University Press; Gierth, M. and Mäser, P. (2007). Potassium transporters in plants – Involvement in K+ acquisition, redistribution and homeostasis. FEBS letters. 581: 2348-2356.
 Guard cells are model systems for
the study of K+ transport
OPEN
 CLOSING


V-ATPase

H+

H+
 The depth and breadth of
V‐PPase
 information available for
A‐
 stomatal guard cells has
H+
 H+
 made them the premier cell
A‐

system in plants for studies
of membrane transport,
H+
 TPK
 signaling, and homeostasis
K+

K+

H+
 K+

TPC

PM‐H+‐ATPase
 GORK

KAT K+

Hills, A., Chen, Z.-H., Amtmann, A., Blatt, M.R. and Lew, V.L. (2012). OnGuard, a computational platform for quantitative kinetic modeling of guard cell physiology. Plant Physiol. 159: 1026-1042 Chen, Z.-H., Hills, A.,
Bätz, U., Amtmann, A., Lew, V.L. and Blatt, M.R. (2012). Systems dynamic modeling of the stomatal guard cell predicts emergent behaviors in transport, signaling, and volume control. Plant Physiology. 159: 1235-1251.
 Potassium homeostasis:
Responses to low K+ availability
Low
K


Membrane
 Hormonal
changes

hyperpolariza+on
 (auxin,
ethylene)


Calcium
 Enhanced root

Direct
effects

signaling
 growth and
Indirect
effects

gravitropic
Transcrip+onal
 responses
More
efficient

induc+on
of
HAK5
K+

uptake
through
K
+
channels

channel


K+ uptake
Adapted from Chérel, I., Lefoulon, C., Boeglin, M. and Sentenac, H. (2013). Molecular mechanisms
involved in plant adaptation to low K+ availability. J. Exp. Bot. 65: 833-848.
 Summary: Potassium uptake,
transport and homeostasis
•  Potassium
is
an
essenIal
macronutrient
required
in

large
amounts

•  Potassium
is
transported
by
channels
and

transporters
which
are
regulated
transcrip+onally
and

post‐transcrip+onally,
by
membrane
voltage
poten+al,

and
signals
such
as
pH,
Ca2+
and
hormones

•  K+
uptake,
transport
and
remobiliza+on
are
regulated

extensively
to
ensure
that
the
plant’s
cri+cal
+ssues

are
preferen+ally
supported

 Sodium
toxicity,
transport
and

tolerance



You
can’t
take
salt
out

of
soil
easily;
once
it
is

there
it
stays
there

To
demonstrate
his
(fake)

madness,
Odysseus
plowed
salt

into
his
field


Colum, P. (1918). The Adventures of Odysseus and the Tale of Troy. Project Gutenberg; USDA, USDA, Peggy Greb; FAO
 Saline soils occur worldwide and
are becoming more abundant
Global
distribuJon
of
salt‐affected
soils


Area
of
salinizaJon


Approximately 7 % of world’s
land area and 30 % of
irrigated land is salt affected

FAO; From: Corbishley, J. and Pearce, D., Growing trees on salt-affected land. ACIAR Impact Assessment Series Report No. 51,
July 2007; See also Munns, R. and Tester, M. (2008). Mechanisms of salinity tolerance. Annu. Rev. Plant Biol. 59: 651-681..
 Coastal and inland soils become
saline for different reasons
Sea
spray

Coastal
areas:
Saline
soils

occur
due
to
intrusion
of

seawater
aggravated
by
 Soil

Coastal
 France
 storms,
rising
sea
levels
and

Ground

lowering
water
tables
 water


Seawater

Spain


Soil
 Rising
sea

Inland
areas:

 Lowering level

ground
Low
rainfall
and
high
rates
of

water table
evotranspira+on

Seawater


European Soil Portal

 Melting land ice is raising sea levels and
threatening agricultural lands
Low‐lying
countries
such
as
Bangladesh
are

par+cularly
vulnerable
–here
a
farmer
surveys

the
damage
caused
by
a
cyclone

Sea
level
change
(cm)


Sea
levels
are
expected

to
be


>1
m
higher
by
2100


Reprinted from Nicholls, R.J. and Cazenave, A. (2010). Sea-level rise and its impact on coastal zones. Science. 328: 1517-1520 by permission of AAAS; Vermeer, M. and Rahmstorf, S. (2009). Global
sea level linked to global temperature. Proc. Natl. Acad. Sci. 106: 21527-21532.See also Cazenave, A. and Llovel, W. (2010). Contemporary sea level rise. Annu. Rev. Marine Sci. 2: 145-173. IRRI
 The San Francisco Bay and Delta
are becoming increasingly salty
Increased
evapora+on

causes
river
water
also

to
be
more
salty


Sacramento

River


San

Joaquin

San Francisco, River
 •  As less water flows through rivers into the
California delta and bay, salty water moves inland
•  Decreased river flows are caused by drought
(less rain) and increased diversion of water to
other parts of the state

DeltaModelingAssociates
Inland, many soils lie above ancient
deep salt deposits that can move up
Clearing
naJve
vegetaJon
oHen
leads
to
soil
salinizaJon


Salty
 Salty

water
 water

Salty
water


Department of Agriculture and Rural Affairs (1980). ‘Managing Salinity: Ensuring a Farming Future’. The State of Victoria
 Irrigation also contributes to soil
salinity by mobilizing deep salts
Rain
 Rain
 Without
irriga+on

Evapora+on
 rainwater
does
not

penetrate
below
the

rootzone


Excessive
irriga+on

penetrates
into
deeper,
salty

soils,
dissolves
the
salts
and

draws
them
upwards
into
the

rootzone


Salt


(dissolved)

 How can we address the problems
caused by soil salinization?
Avoid
adding
to
the

Learn
about
salt

problem
by
be\er

tolerance
from

management
of
fragile

naturally
salt‐
soil
systems

tolerant
species

Areas of concern (halophytes)

Salicornia
europaea


Iden+fy
halophytes
 Arthrocnemum

that
can
be
used
as
 macrostachyum

food
or
energy
crops


Iden+fy
responses

Chenopodium
 Thinopyrum
 to
salt
stress
in

quinoa
 ponIcum

salt‐sensi+ve

Study
salt‐tolerant
rela+ves
of
 species

crop
plants
 (glycophytes)

Introduce
salinity‐tolerance

Munns, R., James, R.A., Xu, B., Athman, A., Conn, S.J., Jordans, C.,
Byrt, C.S., Hare, R.A., Tyerman, S.D., Tester, M., Plett, D. and traits
into
crop
plants
through
 Geng, Y., Wu, R., Wee, C.W., Xie, F., Wei, X., Chan, P.M.Y.,
Gilliham, M. (2012). Wheat grain yield on saline soils is improved by Tham, C., Duan, L. and Dinneny, J.R. (2013). A spatio-temporal
an ancestral Na+ transporter gene. Nat Biotech. 30: 360-364. breeding
and
engineering
 understanding of growth regulation during the salt stress response
CSIRO; The State of Victoria; Maurice Chédel; Marco Schmidt in Arabidopsis. Plant Cell. 25: 2132-2154.
 Plant species have a broad range
of salinity tolerances

Saltbush
(Atriplex
amnicola)
is
a

halophyte
that
can
tolerate
very

salty
soil

Q. Can we identify and
exploit the mechanistic
basis of increased
salinity tolerance?
A. YES!

Arabidopsis
and

rice
are
quite

sensi+ve

Reprinted by permission of Annual Reviews from Munns, R. and Tester, M. (2008). Mechanisms of salinity tolerance. Annu. Rev. Plant Biol. 59: 651-681.
 Mechanisms of sodium toxicity and
tolerance
SALINITY
STRESS


Ionic
stress:

 OsmoJc
stress

K+
deficiency
/
excess

Na+
influx
 Oxida+ve

stress
 InhibiJon
of:



water
uptake,


InhibiJon
of:



growth,




enzyme
ac6vity,




protein
synthesis,
 Detoxifica+on
 

photosynthesis



photosynthesis

 strategies


Leaf
senescence

OsmoJc

Ion
homeostasis:
Na+
 adjustment:

extrusion,
 Accumula+on
of

Na+
exclusion,

 solutes

Na+
compartmenta+on

Adapted from Horie, T., Karahara, I. and Katsuhara, M. (2012). Salinity tolerance mechanisms in glycophytes: An overview with the central focus on rice plants. Rice. 5: 11; see also Munns, R. and Tester, M. (2008). Mechanisms of salinity
tolerance. Annu. Rev. Plant Biol. 59: 651-681 and Shabala, S. and Pottosin, I. (2014). Regulation of potassium transport in plants under hostile conditions: implications for abiotic and biotic stress tolerance. Physiol. Plant. 151: 257-279.
 General sodium tolerance strategy:
Keep sodium out of cytosol & shoot
OUT

1.
Keep
Na+
from

Na+

 4.
Extrude
Na+
via

IN
 Na+


salt
glands

entering
plant
/
cells

“OUT”

K+

 5.
Accumulate
K+
to

2.
Pump
out
any
Na Na+

 maintain
a
high
ra+o

+
that
leaks
in

of
K+
to
Na+


3.
Compartmenta+on
of
 Na+

 CompaJble


Na+
in
vacuole
 solutes


6.
Synthesize
compa+ble

solutes
for
osmo+c

balance
 7.
Prevent
Na+

from
moving
into

the
shoot
and

leaves

 Na+ transport and exclusion is an
integral part of Na+ tolerance
As
Na+
becomes
more

Cytosol
 prevalent,
it
is
preferen+ally

Vac.
 sequestered
into
the
vacuole

via
transporters


Na+
can
be
sequestered
in
 Priori6zed

less
essen+al
+ssues
and

excluded
from
growing
and

photosynthe+c
+ssues

Non‐
Priori6zed

Adapted from Amtmann, A., and Leigh, R. (2010). Ion homeostasis. In Abiotic Stress Adaptation in Plants: Physiological, Molecular and Genomic
Foundation, A. Pareek, S.K. Sopory, H.J. Bohnert and Govindjee (eds) (Dordrecht, The Netherlands: Springer), pp. 245 – 262.
 Ion pumps, channels & carriers
contribute to Na+ tolerance

PM-H+-ATPase Na+

ATP
 ADP
+
Pi

H+

SOS1, NHX8 H+

SOS1
H+
 H+

Na+
 ATP
 ADP+
Pi


H+
 H+

PP
 2
x
Pi

HKT Na+


NSCC Na+
 Na+


NHX H+-PPase VH+-ATPase

See Maathuis, F.J.M. (2014). Sodium in plants: perception, signalling, and regulation of sodium fluxes. J. Exp. Bot. 65: 849-858.
 HKTs have essential roles in salt
exclusion and salinity tolerance
Expression of HKT1 in the cells
HKT
stands
for
“high
 surrounding xylem in the root helps
affinity
K+
transport”
but

prevent Na+ from reaching the
they
also
contribute
to
Na
+
transport
 photosynthetic cells in the shoot

Xylem
 Leaf

Expression
pa\ern
in
rice
root;

blue
indicates
gene
expression


Root
 Na+



Adapted from Davenport, R.J., MuÑOz-Mayor, A., Jha, D., Essah, P.A., Rus, A.N.A. and Tester, M. (2007). The Na+ transporter AtHKT1;1 controls retrieval of Na+ from the xylem
in Arabidopsis. Plant Cell Environ. 30: 497-507. Reprinted by permission from Macmillan Publishers Ltd from Ren, Z.-H., Gao, J.-P., Li, L.-G., Cai, X.-L., Huang, W., Chao, D.-Y.,
Zhu, M.-Z., Wang, Z.-Y., Luan, S. and Lin, H.-X. (2005). A rice quantitative trait locus for salt tolerance encodes a sodium transporter. Nat Genet. 37: 1141-1146.
 HKT1 expression level and activity
is correlated with Na+-tolerance
Loss
of
func+on
=

more
salt
sensi+ve


Salt-tolerance in varieties of
rice, wheat and barley has
been genetically mapped to
variation in HKT activity Gain
of
func+on
=

more
salt
tolerant

Mäser, P., Eckelman, B., Vaidyanathan, R., Horie, T., Fairbairn, D.J., Kubo, M., Yamagami, M., Yamaguchi, K., Nishimura, M., Uozumi, N., Robertson, W., Sussman, M.R. and Schroeder,
J.I. (2002). Altered shoot/root Na+ distribution and bifurcating salt sensitivity in Arabidopsis by genetic disruption of the Na+ transporter AtHKT1. FEBS letters. 531: 157-161.
 Monocots have two types of HKTs
with different functions
Type
1

 Subfamily
1
 Subfamily
2

Retrieval
of
Na+
from

transpira+on
stream

Found
in
all
plants
 Root


Na+


Xylem
 Leaf


K+



Type
2


Postulated
role
in
nutri+onal
Na+

Root
 Na+

 uptake,
when
[K+]
very
low

(ac+vity
suppressed
by
K+)

Only
present
in
monocots

Reprinted from Véry, A.-A., Nieves-Cordones, M., Daly, M., Khan, I., Fizames, C. and Sentenac, H. (2014). Molecular biology of K+ transport across the plant cell membrane: What do we learn from comparison
between plant species? J. Plant Physiol. 171: 748-769 with permission from Elsevier. See also Horie, T., Costa, A., Kim, T.H., Han, M.J., Horie, R., Leung, H.-Y., Miyao, A., Hirochika, H., An, G. and Schroeder, J.I.
(2007). Rice OsHKT2;1 transporter mediates large Na+ influx component into K+-starved roots for growth. EMBO J. 26: 3003-3014.
 NHX (Sodium / proton exchangers) are
part of the CPA family

Arabidopsis:
8
NHX
transporters


AtNHX
1
–
4
Vacuole


AtNHX
5
–
6
Endosome


AtNHX
7
(SOS1)
–
8
PM
 CPA = Cation / proton antiporter
Chérel, I., Lefoulon, C., Boeglin, M. and Sentenac, H. (2013). Molecular mechanisms involved in plant adaptation to low K+ availability. J. Exp. Bot. 65: 833-848. Gierth, M. and Mäser, P.
(2007). Potassium transporters in plants – Involvement in K+ acquisition, redistribution and homeostasis. FEBS letters. 581: 2348-2356. Chanroj, S., Wang, G., Venema, K., Zhang, M.W.,
Dalwiche, C.F., and Sze, H. (2012). Conserved and diversified gene families of monovalent cation / H+ antiporters from algae to flowering plants. Front. Plant Sci. 3: 25.
Loss of function of SOS1 makes
plants “salt overly sensitive”

H+

Na+


SOS1, aka NHX7, is a plasma-

membrane localized Na+/H+ exchanger

It
has
an
auto‐inhibitory
domain
that

can
be
phosphorylated
to
ac+vate
the

protein
under
salinity
stress


Wu, S.J., Ding, L. and Zhu, J.K. (1996). SOS1, a Genetic Locus Essential for Salt Tolerance and Potassium Acquisition. Plant Cell. 8: 617-627. Shi, H., Ishitani, M., Kim,
C. and Zhu, J.-K. (2000). The Arabidopsis thaliana salt tolerance gene SOS1 encodes a putative Na+/H+ antiporter. Proc. Natl. Acad. Sci. USA. 97: 6896-6901.
 NHXs roles include Na+, K+ and H+
transport and homeostasis
Outward
Na+
across
PM

 Na+
sequestraJon
in
vacuole


RegulaJon
of
pH
of

endocyJc
compartments

for
proper
protein

sorJng
and
modificaJon


Bassil, E., Coku, A. and Blumwald, E. (2012). Cellular ion homeostasis: emerging roles of intracellular NHX Na+/H+ antiporters in plant growth and development. J. Exp. Bot. 63: 5727-5740, by permission of Oxford University Press.
 Iden+fica+on
of
salt
tolerance
in

halophytes
and
crop
rela+ves

Most
sensi6ve
 Most
tolerant


Glycophytes
 Halophytes


Arthrocnemum

Wheat
–
intermediate
sensi+vity
 macrostachyum

(Tri6cum
aes6vum)



Salinity tolerance is a complex

Chick
pea
 trait and plants have a wide range
(Cicer
arie6num)
 of salinity tolerances
Halophytes are often defined as
having the ability to complete their
lifecycle on >200 mM NaCl Salicornia
spp.


See Flowers, T.J., Galal, H.K. and Bromham, L. (2010). Evolution of halophytes: multiple origins of salt tolerance in land plants.
Functional Plant Biology. 37: 604-612. Photo credits: Sanjay Acharya; Z. Hell; Fritz Geller-Grimm; Javier Martin, H2O
 Salt tolerance has evolved repeatedly
and independently
RED
indicates
order
that

includes
some
halophytes

All
vascular
plants


200 salt tolerant species

within the grasses, from
~76 independent events

Flowers, T.J., Galal, H.K. and Bromham, L. (2010). Evolution of halophytes: multiple origins of salt tolerance
in land plants. Funct. Plant Biol. 37: 604-612; Bennett, T.H., Flowers, T.J. and Bromham, L. (2013). Repeated
evolution of salt-tolerance in grasses. Biol. Lett. 9: 20130029, by permission of the Royal Society.
Halophytes can be grown on saline
soils for food and fodder
The
genus
Atriplex
includes
many

edible
halophytes
and
is
being
grown

for
fodder
in
Australia


Palmer’s
grass
(or
nipa
grass;
Dis6chlis

palmeri)
grows
in
+dal
marshes
of
the
Gulf
of

California
and
was
a
food
grain
eaten
by
the

indigenous
people
of
the
region


USA

Atriplex
nummularia

(old
man
saltbush)

Mex


Image credits: M. Fagg, Australian National Botanic Gardens; Arizona State University. See Glenn, E.P., Anday, T., Chaturvedi, R., Martinez-Garcia, R., Pearlstein, S., Soliz, D., Nelson, S.G. and
Felger, R.S. (2013). Three halophytes for saline-water agriculture: An oilseed, a forage and a grain crop. Env. Exp. Bot. 92: 110-121;Flowers, T.J. and Colmer, T.D. (2008). Salinity tolerance in
halophytes*. New Phytol. 179: 945-963. Shabala, S. (2013). Learning from halophytes: Physiological basis and strategies to improve abiotic stress tolerance in crops. Ann. Bot. 112: 1209-1221.
 Quinoa is a facultative halophyte and a
popular food grain
Young
leaves
(leM)
extrude
salt
into
salt
bladders,

Quinoa is also a older
leaves
(right)
store
it
in
vacuoles


useful model for

studies of salinity
tolerance

Control

Control

Salt

Salt


Quinoa
(Chenopodium
quinoa)
evolved
in
the
 Vacuolar
sodium
channel
ac+vity
is
decreased
in
old

leaves

Andes
and
can
tolerate
saline
soils

 (right)
grown
under
salinity;
no
salinity
effect
is
observed
in

young
leaves

Bonales-Alatorre, E., Shabala, S., Chen, Z.-H. and Pottosin, I. (2013). Reduced tonoplast fast-activating and slow-activating channel
activity is essential for conferring salinity tolerance in a facultative halophyte, Quinoa. Plant Physiol. 162: 940-952. Maurice Chédel
 Models for salt tolerance:
Eutrema spp. (salt /saltwater cress)

Strategies
for
salt
tolerance
include

expansion
of
several
gene
families

(HKT,
AVP)
and
lower
accumula+on
of

Na+
in
the
shoot
as
compared
to

Arabidopsis..



Arabidopsis
thaliana


Higher
selec+vity
for

K+
than
Na+
uptake
in

the
root


(Previously
known
as

Thellungiella
halophila

or
the
related


Thellungiella

salsuginea)

Reprinted from Amtmann, A. (2009). Learning from evolution: Thellungiella generates new knowledge on essential and
critical components of abiotic stress tolerance in plants. Mol. Plant. 2: 3-12 by permission of Oxford University Press.
 Breeding
and
engineering
for
salt

tolerance


Salt
tolerance
can
be

ajributed
to
three

non‐exclusive

mechanisms



Salinity
tolerance can
be enhanced by
breeding or
engineering

Reprinted from Roy, S.J., Negrão, S. and Tester, M. (2014). Salt resistant crop plants. Curr. Opin. Biotech. 26: 115-124.
Wheat yield on saline soils improved by an
ancestral Na+ transporter gene
A pair of genes derived from a
relative of wheat confers enhanced
salinity tolerance

Because
these
species
are

closely
related,
the
genes
can

be
introduced
into
cul+vated

wheat
without
using
GM

methods


Durum
wheat
carrying
salt‐tolerance

genes

Tetraploid
pasta
 Hexaploid
bread

wheat
 wheat


Huang, S., Spielmeyer, W., Lagudah, E.S. and Munns, R. (2008). Comparative mapping of HKT genes in wheat, barley, and rice, key determinants
of Na+ transport, and salt tolerance. J. Exp. Bot. 59: 927-937 by permission of Oxford University Press; Credit: Dr Richard James, CSIRO
 Nax1 and Nax2 exclude Na+ from leaf
blades by removal from xylem
When
expressed
in
Xenopus

oocytes,
the
transporters

conduct
Na+
but
not
K+


Nax1

Nax2

In plants, Nax1 and Nax2
pump Na+ into the cells
surrounding the xylem so it
does not reach the leaf blade

Reprinted by permission from Macmillan Publishers Ltd from Munns, R., et al and Gilliham, M. (2012). Wheat grain yield on saline soils is improved by an ancestral Na+ transporter gene. Nat.
Biotech. 30: 360-364. Schroeder, J.I., et al and and Sanders, D. (2013). Using membrane transporters to improve crops for sustainable food production. Nature. 497: 60-66. Huang, S., Spielmeyer,
W., Lagudah, E.S. and Munns, R. (2008). Comparative mapping of HKT genes in wheat, barley, and rice, key determinants of Na+ transport, and salt tolerance. J. Exp. Bot. 59: 927-937.
 The candidate gene approach has
had some success
Transgenic
plants
carrying
vacuolar
NHXs,
vacuolar
H+‐
PPases
and
plasma
membrane
NHXs
have
demonstrated

enhanced
salinity
tolerance


PM-H+-ATPase Na+

ATP
 ADP
+
Pi

H+

H+

SOS1 Enhanced
ROS

ATP

H+

ADP+
Pi

detoxifica+on
and

H+
 PP

H+

2
x
Pi
 synthesis
of

HKT Na+

compa+ble
solutes
is

NSCC Na+
 Na+
 also
correlated
with

NHX H+-PPase V-H+-ATPase
enhanced
salinity

tolerance


See for example Roy, S.J., Negrão, S. and Tester, M. (2014). Salt resistant crop plants. Curr. Opin. Biotechnology. 26: 115-124; Gaxiola, R.A., Li, J., Undurraga, S., Dang, L.M., Allen,
G.J., Alper, S.L. and Fink, G.R. (2001). Drought- and salt-tolerant plants result from overexpression of the AVP1 H+-pump. Proc. Natl. Acad. Sci. USA 98: 11444-11449; Apse, M.P.,
Aharon, G.S., Snedden, W.A. and Blumwald, E. (1999). Salt tolerance conferred by overexpression of a vacuolar Na+/H+ antiport in Arabidopsis. Science. 285: 1256-1258.
The
intersec+on
of
potassium
nutri+on

and
sodium
toxicity

K+
uptake


K+ext
 Na+ext


Na+
uptake


Na+
and
K+
interfere
with

each
other’s
uptake


When
barley
plants
are
grown

on
200
mM
NaCl,
they

accumulate
Na+
at
the
expense

of
K+
in
their
leaves


Cuin, T.A., Miller, A.J., Laurie, S.A. and Leigh, R.A. (2003). Potassium activities in cell compartments of salt‐grown barley leaves. J. Exp. Bot. 54: 657-661 with permission from Oxford University Press.
 As [Na+]ext increases and enters the cell, K+ is
driven out
1.
Steep
concentra+on

gradient
for
Na+
 Na+

Na+

2.
Na+
leaks
in
through
 Non-selective
cation channel Na+

NSCCs


3.
Increased
nega+ve
 Membrane
 K+


charge
within;
 depolarizes

depolarized
membrane


K+
 Furthermore,
Na
4.
K+
driven
out
through
 +
directly

Kv
channel

ATP
 competes
with
K
+
for
low‐
and

H+

Some
salt‐tolerant
plants

maintain
 high‐affinity

ADP
+
Pi

elevated
K+
by
higher
ac+vity
of
PM
 transporters


H+‐ATPase


Chen, Z., Pottosin, I.I., Cuin, T.A., Fuglsang, A.T., Tester, M., Jha, D., Zepeda-Jazo, I., Zhou, M., Palmgren, M.G., Newman, I.A. and Shabala, S. (2007). Root plasma membrane transporters
controlling K+/Na+ homeostasis in salt-stressed barley. Plant Physiol. 145: 1714-1725; Shabala, S. and Cuin, T.A. (2008). Potassium transport and plant salt tolerance. Physiol. Plant. 133: 651-669.
 Interaction between K+ nutrition and Na+ toxicity

Cytosol

Vac.


K+
/
Na+
raIo


Plants must Priori6zed


coordinate the
actions of K+ and Na+
transporters to Non‐
maintain a high ratio Priori6zed

of K+ to Na+ in
prioritized tissues
K+
/
Na+
raIo

Adapted from Amtmann, A., and Leigh, R. (2010). Ion homeostasis. In Abiotic Stress Adaptation in Plants: Physiological, Molecular and Genomic
Foundation, A. Pareek, S.K. Sopory, H.J. Bohnert and Govindjee (eds) (Dordrecht, The Netherlands: Springer), pp. 245 – 262.
Salinity tolerance: Summary

•  Saline
soils
are
detrimental
to
plants
and
are

widespread

•  Sodium
toxicity
is
primarily
due
to
interfering

with
K+
nutri+on

•  Sodium
tolerance
depends
on
exclusion,

extrusion
an
sequestra+on

•  Breeding
and
engineering
for
salinity

tolerance
have
had
mixed
success
so
far

 Summary and ongoing research
•  Nutrient
uptake
is
extremely
energe+cally
demanding

•  Proton
moIve
force
generated
by
proton
pumps
is
essen+al

for
nutrient
uptake

•  Dozens
of
membrane
transporters
are
involved
in
uptake,

alloca+on
and
homeostasis
of
mineral
nutrients


•  Most
plants
require
a
high
cytosolic
raIo
of
K+
to
Na+

•  Plants
require
large
amounts
of
potassium
for
op+mal
growth


NO3‐

NO3‐

•  Sodium
toxicity
is
a
real
and
growing
problem

K+

•  The
mechanisms
of
sodium
tolerance
are
being

K+
 iden+fied
and
exploited
for
plant
breeding

PO43‐

PO43‐

PO43‐