Chapter 2

Chapter 2
Ion Uptake Mechanisms of Individual Cells

and Roots: Short-distance Transport
Philip J. White
Scottish Crop Research Institute, Invergowrie, Dundee, UK
SUMMARY
cases nutrient solutions, can contain high concentrations of
The uptake of nutrients by higher plants is characterized by selec- mineral elements not needed for plant growth, or that are
tivity of transport and accumulation in specific tissues, cells or potentially harmful to plants. The mechanisms by which
subcellular compartments. These characteristics are genetically plants accumulate nutrients must therefore be selective.
determined and can differ both between and within plant spe- This selectivity can be demonstrated particularly well in
cies. This chapter reviews the environmental, physiological and algal cells (Table 2.1), where the external and vacuolar
developmental factors that affect the entry of nutrients into the (cell sap) solutions are separated by only two membranes:
extracellular space (apoplasm) of roots, their transport across the plasma membrane and the tonoplast.
the plasma membrane and tonoplast of root cells, and the path- In Nitella, the concentrations of K, Na, Ca, and Cl ions
ways of their movement to the xylem. It describes the structure are higher in the cell sap than in the pond water, but the
and composition of cellular membranes, the electrochemical
concentration ratio differs considerably between the ions.
gradients that determine the energetics of solute transport across
membranes, and the mechanisms involved and the genetic iden-
By contrast, in Valonia growing in seawater, only K is more
tity of the proteins that facilitate the transport of nutrients across concentrated in the cell sap, whereas the Na and Ca concen-
the plasma membrane and tonoplast of plant cells. The overriding trations are lower in the cell sap than in the seawater.
influence of plant nutritional status on the expression of mecha- Selective ion uptake is also a typical feature of higher
nisms by which roots acquire nutrients is emphasized. plants. When plants are grown in a nutrient solution of
limited volume, the external concentrations of ions change
with time (Table 2.2). The concentrations of K, P and
2.1 GENERAL
nitrate decline markedly, whereas those of Na and sulphate
As a rule, there is a great discrepancy between the con- can even increase, indicating that water is taken up faster
centrations of mineral nutrients in the soil and the nutri- than either of these two ions. Uptake rates, especially for K
ent requirements of plants. Furthermore, soil, and in some and Ca, differ between plant species (e.g., maize and bean,
TABLE 2.1 Relationship between ion concentrations in the substrate and in the cell sap of Nitella and Valonia
Nitella concentration (mM) Valonia concentration (mM)
A B Ratio A B Ratio
Ion Pond water Cell sap B/A Seawater Cell sap B/A
Potassium 0.05 54 1080 12 500 42
Sodium 0.22 10 45 498 90 0.18
Calcium 0.78 10 13 12 2 0.17
Chloride 0.93 91 98 580 597 1
Modified from Hoagland (1948).
Marschner’s Mineral Nutrition of Higher Plants. DOI: 10.1016/B978-0-12-384905-2.00002-9

© 2012 Elsevier Ltd. All rights reserved. 7
8 PART | I Nutritional Physiology
TABLE 2.2 Changes in the ion concentration of the external (nutrient) solution and in the root
sap of maize and bean
External concentration (mM)
Concentration in root
After 4 daysa sap (mM)
Ion Initial Maize Bean Maize Bean
Potassium 2.00 0.14 0.67 160 84
Calcium 1.00 0.94 0.59 3 10
Sodium 0.32 0.51 0.58 0.6 6
Phosphate 0.25 0.06 0.09 6 12
Nitrate 2.00 0.13 0.07 38 35
Sulfate 0.67 0.61 0.81 14 6
a
No replacement of water lost through transpiration.
FIGURE 2.1 Cross-section of two rhizodermal cells of a maize root. V, vacuole; C, cytoplasm; W, cell wall, E, external solution. Courtesy of
C. Hecht-Buchholz.
Table 2.2). The concentrations of ions in the root sap are it is necessary to follow the pathway of solutes (ions,
generally higher than those in the nutrient solution; this is charged and uncharged molecules) from the external solu-
most evident in the case of K, nitrate and phosphate. tion through the cell wall and the plasma membrane into
The results obtained from both lower and higher plants the cytoplasm and vacuoles of plant cells.
demonstrate that ion uptake is characterized by:
1. Selectivity. Certain mineral elements are taken up pref- 2.2 PATHWAY OF SOLUTES FROM THE
erentially, while others are discriminated against or EXTERNAL SOLUTION INTO ROOT
almost excluded. CELLS
2. Accumulation. The concentration of elements can be
much higher in cell sap than in the external solution. 2.2.1 Influx to the Apoplasm
3. Genotype. There are distinct differences between plant
Movement of low-molecular-weight solutes (e.g., ions,
species in their ion uptake characteristics.
organic acids, amino acids, sugars) from the external solu-
These observations lead to many questions. In particu- tion through the walls of individual root cells (the free
lar, how do individual cells and higher plants regulate the space) is a non-metabolic, passive process, driven by dif-
uptake of ions both to satisfy plant demand and to avoid fusion or mass flow (Fig. 2.1). Nevertheless, cell walls
ion toxicities? To understand the regulation of ion uptake can interact with solutes and, thereby, facilitate or restrict
Chapter | 2 Ion Uptake Mechanisms of Individual Cells and Roots: Short-distance Transport 9
TABLE 2.3 Diameter of cell wall pores and hydrated

cations Indiffusible
anions
Diameter (nm)
Rhizodermal cell wall (maize; Fig. 2.1) 500–3,000
Cortical cell wall (maize) 100–200 Cation
Anion
Pores in cell wall 5.0
Sucrose 1.0 Macropore
WFS
DFS
Hydrated ions Micropore
K 0.66
FIGURE 2.2 Schematic diagram of the pore system of the apparent free
Ca2 0.82 space. DFS, Donnan free space; WFS, water free space.
passage across the root and uptake across the plasma mem- TABLE 2.4 Cation exchange capacity of root dry
brane of individual cells. matter of different plant species
The primary cell wall consists of a network of cellu- Cation exchange capacity
lose, accounting for about 15–30% of its dry weight, cross- Plant species (mmol (100 g)1 dw)
linking glycans (generally xyloglucans in Type I walls,
Wheat 23
but in the Type II walls of commelinoid monocotyledons
mostly gluconoarabinoxylans) and glycoproteins, all Maize 29
embedded in a pectin matrix (Carpita and McCann, 2000). Bean 54
Type I cell walls contain more pectin than Type II cell
Tomato 62
walls. Both Ca and B are also integral components of cell
Based on Keller and Deuel (1957).
walls, which can be additionally impregnated with Si. The
cell wall network contains pores, the so-called interfibril-
lar and intermicellar spaces, which differ in size. For root
hair cells of radish, a maximum diameter of 3.5 to 3.8 nm by typical Donnan distributions. Trivalent cations, such
(35–38 Å) has been calculated and maximum diameters for as Al3, bind more strongly than divalent cations, such as
the pores in plant cell walls are generally in the range of Ca2, which bind more strongly than monovalent cations,
5.0 nm (Table 2.3). The diameters of hydrated ions, such such as K. Plant species differ considerably in their cat-
as K and Ca2, are small in comparison. Therefore, ion exchange capacity (CEC), that is, in the number of cat-
the pores themselves would not be expected to offer any ion exchange sites (fixed anions; R·COO), located in cell
restriction to ion movement through the cell wall. walls (Table 2.4).
In contrast to nutrients and low-molecular-weight organic As a rule, the CEC of dicotyledonous species is
solutes, the movement of high-molecular-weight solutes greater than that of monocotyledonous species (White
(e.g., metal chelates, fulvic acids and toxins) or viruses and and Broadley, 2003). As the external pH decreases, the
other pathogens through one cell walls is severely restricted effective CEC is reduced, particularly in monocotyledo-
by the diameter of the pores. nous species (Allan and Jarrell, 1989) as protons occupy
A variable proportion of the pectins in cell walls con- an increasing proportion of the cation binding sites.
sist of polygalacturonic acid, originating mainly from Because of apoplastic barriers within the root, such as the
the middle lamella. Accordingly, their carboxylic groups Casparian band of the endodermis and exodermis, only
(R.COO) act as cation exchangers in the cell wall con- part of the AFS is directly accessible to cations from the
tinuum of roots and other plant tissue, the so-called apo- external solution. Exchange adsorption of cations in the
plasm. In roots, cations from the external solution can apoplasmic AFS is not a prerequisite for ion uptake across
accumulate in the free space, whereas anions are repelled. the plasma membrane or for the movement of ions within
Hope and Stevens (1952) introduced the term appar- the apoplasm. However, fixed negative charges in the AFS
ent free space (AFS). This comprises the water free space can influence both the absolute and relative concentrations
(WFS), which is freely accessible to ions and charged and of cations in the apoplasm, especially when roots grow in
uncharged molecules, and the Donnan free space (DFS), dilute solutions (White and Broadley, 2003). Thus, root
where cation exchange and anion repulsion take place (Fig. CEC can affect the rate and selectivity of ion influx into
2.2). Ion distributions within the DFS are characterized root cells and apoplasmic ion movements indirectly. It
has been speculated that this could account for the posi- 4 I
45Ca 42K
µmol (g root fresh wt)−1

tive correlation between root CEC and the ratio of Ca2 to I
K concentration in different plant species. Alternatively, 3
this correlation could simply reflect the predominant accu-
mulation of Ca2 in the apoplasm and of K in the vacu- 2 E
oles of plant cells (White and Broadley, 2003; White and
Karley, 2010). 1
E
The apoplasmic AFS can also serve as a transient stor-
age pool for essential mineral elements such as iron and 0
zinc which can be mobilized, for example, by specific root 0 10 30 50 0 10 30 50
exudates such as phytosiderophores, and subsequently Time (min) Time (min)
translocated to the shoots (Zhang et al., 1991b, c; Cesco FIGURE 2.3 Time course of influx (I) and efflux (E) of 45Ca and 42K to
et al., 2002; Liu et al., 2010). For iron, the size of this stor- isolated barley roots. After 30 min (arrow) some of the roots were trans-
age pool possibly contributes to genotypic differences in ferred to solutions with no labelled Ca2 and K. The proportion of the
sensitivity to iron deficiency in soybean (Longnecker and exchangeable fraction in the apparent free space is calculated by extrapo-
Welch, 1990). In addition, the entry of elements supplied lation to zero time.
to the root in excess of plant demand, such as Ca in calcar-
eous soils, can be restricted by precipitation as insoluble
salts. TABLE 2.5 Reflection coefficients (δ) of some non
electrolytes at the cell membranes of Valonia utricularis
2.2.2 Passage into the Cytoplasm Molecular
Compound δa radius (nm)
Despite some selectivity for cation binding in the cell wall,
Raffinose 1.00 0.61
the main site of selectivity in the uptake of cations and
anions, as well as solutes in general, is the plasma mem- Sucrose 1.00 0.53
brane of individual cells. The lipid bilayer of the plasma Glucose 0.95 0.44
membrane prevents the indiscriminate movement of ions
Glycerol 0.81 0.27
and large polar molecules from the apoplasm into the cyto-
plasm (influx) and from the cytoplasm into the apoplasm Urea 0.76 0.20
(efflux). Integral membrane proteins facilitate the selec- Based on Zimmermann and Steudle (1970).
tive transport of solutes across the plasma membrane. It a
1.00 indicates that the membranes are impermeable to the solute; 0.00
indicates that the membranes are freely permeable to the solute.
can be readily demonstrated that the plasma membrane is
a selective barrier to the uptake of ions. For example, when
barley plants are placed in a nutrient solution containing
Ca2 (45Ca) and K (42K), most of the 45Ca accumulated provide the unique transport properties required for the
in the roots in the first 30 min is still readily exchangeable function of each compartment. Before the mechanisms of
and is almost certainly located in the apoplasmic AFS (Fig. solute transport across membranes are discussed in greater
2.3). By contrast, only a minor fraction of the 42K is read- detail (Sections 2.4 and 2.5), some fundamental aspects
ily exchangeable after this period, most of the 42K having of the composition and structure of biological membranes
been transported across the plasma membranes of root will be described.
cells. Furthermore, in most mature plant cells, the vacuole
comprises more than 80–90% of the cell volume, acting as 2.3 COMPOSITION OF BIOLOGICAL
central storage compartment for ions and solutes (e.g., sug-
ars and secondary metabolites), and it is likely that the 42K
MEMBRANES
is already sequestered within this cellular compartment. The capacity of plant cell membranes to regulate solute
Within the plant cell, membranes with contrasting uptake has fascinated botanists since the nineteenth cen-
lipid and protein composition separate the various cellu- tury. By the early years of the twentieth century some
lar compartments. These membranes include the tonoplast basic facts of solute permeation across biological mem-
(vacuolar membrane), the endoplasmic reticulum (ER), branes had been established, such as the inverse relation-
the Golgi apparatus, the nuclear membrane and the mem- ship between the diameter of uncharged molecules and
branes surrounding vesicles, mitochondria and plastids the rates at which they permeate membranes (Table 2.5).
(Staehelin and Newcomb, 2000). For these membranes, High-molecular-weight organic solutes such as polyeth-
the lipid bilayer provides the barrier to solute movement, yleneglycol are not taken up by cells and can be used at
and proteins facilitate the selective transport of solutes to high external concentrations as osmotica to induce water
(A) Transmembrane (B) Lipid linked (C) Protein attached
NH2
Lipid
bilayer
COOH
FIGURE 2.4 Protein associations with biological membranes. Integral transmembrane proteins extend through the lipid bilayer in α-helical or β-sheet
structures. Peripheral proteins are attached to the membrane either by covalently attached lipid groups or through interactions with integral membrane
proteins. Based on a figure from Essential Biology of the cell by Bruce Alberts et al. (1998). Reproduced by permission of Garland Science/Taylor &
Francis Books, Inc.
The most abundant membrane lipids are (i) phospholi-

TABLE 2.6 Lipid and fatty acid composition of the pids, in which the hydrophilic headgroup is linked to the
plasma membrane and tonoplast of mung bean hydrophobic tail by a phosphate group, (ii) sterols, which
Plasma membrane Tonoplast (µmol are based around a four-ring structure, and (iii) glycolip-
Lipids (µmol mg1 protein) mg1 protein) ids, which have sugars as their hydrophilic headgroup (Fig.
2.5). Common plant phospholipids are phosphatidyl ser-
Phospholipids 1.29 1.93
ine, phosphatidyl ethanolamine, phosphatidyl choline,
Sterols 1.15 1.05 phosphatidyl inositol and diphosphatidyl glycerol. Plasma
Glycolipids 0.20 0.80 membranes and mitochondria are enriched in phosphati-
dyl inositol and diphosphatidyl glycerol, respectively. The
Fatty acid composition of the phospholipids fatty acid moiety in phospholipids varies in both chain
Plasma Tonoplast length and number of double bonds, but is often palmitic
Chain Melting membrane (% of (length:double-bonds, 16:0), stearic (18:0), oleic (18:1),
Fatty acid length point (°C) (% of total) total) linoleic (18:2) or linolenic (18:3) acid (Table 2.6). Major
Palmitic acid C16 62.8 35 39 plant sterols include campesterol, sitosterol and stigmas-
Stearic acid C18 70.1 6 6
terol. The sterol content of the ER is low, but sterols can
make up more than 30% of the total lipids in the plasma
Oleic acid C18:1a 13.0 9 9 membrane and tonoplast (Table 2.6). Most glycolipids
Linoleic acid C18:2a 5.5 21 22 are found in the chloroplast (Hölzl and Dörmann, 2007),
a where the thylakoid membrane is predominantly com-
Linolenic acid C18:3 11.1 19 20
posed of monogalatosyldiacyglycerol (MGDG), together
Others – – 10 4 with digalatosyldiacyglycerol (DGDG) and the sulpho-
Based on Yoshida and Uemura (1986). lipid, sulphoquinovosyldiacylglycerol (SQDG).
a
Numeral to the right of the colon indicates the number of double
bounds.
Although the lipid bilayer provides the basic structure of
the membrane and forms a permeability barrier, most bio-
logical functions of membranes are performed by proteins.
The membrane surrounding each cellular compartment has
deficiency (drought stress) in plants. However, some different types of proteins reflecting the particular func-
hydrophobic molecules penetrate membranes much faster tion of that membrane. Membrane proteins function (i) to
than would be predicted on the basis of their size, which is anchor the membrane to the cytoskeleton and/or cell wall,
presumably related to their ability to partition into the lipid (ii) as receptors/transducers for compartmentalized signals,
bilayer. (iii) as enzymes for specific reactions, such as energy trans-
Biological membranes are typically composed of a duction processes in mitochondria and chloroplasts, and (iv)
lipid bilayer and associated proteins (Fig. 2.4). However, to transport specific solutes across membranes.
membrane composition is sensitive to environmental con- There are several ways by which proteins can be asso-
ditions, and the relative abundance, and types, of both ciated with the lipid bilayer (Staehelin and Newcomb,
lipids and proteins in membranes surrounding cellular 2000). Many membrane proteins extend through the
compartments differ (Table 2.6). The lipids in cell mem- bilayer (Fig. 2.4). These integral transmembrane proteins
branes have hydrophilic headgroups and hydrophobic tails. have both hydrophobic and hydrophilic portions. Their
O
CH2 O P O CH2 CH2 N+(CH3)3
R1 O CH O−
R2 O CH2 Phosphatidylcholine
CH2OH
O
H H H
CH2 O H OH OH
R1 O CH OH H
R2 O CH2
Monogalactosyl diglyceride
O
CH2 S O−
O
H H H O
CH2 O H OH OH
R1 O CH OH H
R2 O CH2
Sulfoquinovosyl diglyceride
C2H5
CH3 CH3
CH3
CH3
CH3
β-Sitosterol
HO
FIGURE 2.5 Chemical structures of selected membrane lipids. Phospholipids are represented by phosphatidylcholine, glycolipids by monogalactosyl
diglyceride and sulphoquinovosyl diglyceride, and sterols by β-sitosterol, respectively.
hydrophobic portions lie within the bilayer, alongside (Westerman and Roddick, 1981); membrane lipid compo-
the hydrophobic tails of the lipid molecules, while their sition changes during exposure to low temperatures (e.g.,
hydrophilic portions extend into the aqueous environment Welti et al., 2002; Penfield, 2008), and DGDG and SQDG
on either side of the membrane. Other membrane proteins can replace phospholipids in membranes of P deficient
are located entirely outside the bilayer. These peripheral plants (Hölzl and Dörmann, 2007; White and Hammond,
proteins are bound to the membrane through lipid groups 2008). Similarly, the composition of root membranes is
attached covalently through prenylation (attachment of the influenced by temperature, salinity and the ionic composi-
isoprenoids farnesyldiphosphate or geranylgeranyldiphos- tion of the external solution (e.g., Cakmak and Marschner,
phate), S-acetylation (attachment of palmitate or stearate) 1988c; White et al., 1990b; Wu et al., 1998; Lindberg
or N-myristolation (Sorek et al., 2009), or are associated et al., 2005; López-Pérez et al., 2009). The changes in
with other membrane proteins through ionic interactions. lipid composition reflect often the adaption of a plant to its
It is thought that lipid modification of membrane proteins environment. For example, membranes of plants growing
also facilitates their subcellular targeting and clustering at low temperatures have more phospholipids with charged
into specific domains. headgroups and shorter fatty acid chains with lower degree
Lipid composition not only differs between cellular of saturation, and greater sterol content than plants grow-
membranes (Table 2.6), plant tissues and plant species ing at higher temperatures (Staehelin and Newcomb, 2000;
(Staehelin and Newcomb, 2000), but is also strongly influ- Wallis and Browse, 2002; Penfield, 2008). Such changes
enced by environmental factors. In leaves, for example, shift the freezing point (i.e., the transition temperature) of
distinct annual variations in sterol concentrations occur membranes to a lower temperature and may therefore be
2.4 SOLUTE TRANSPORT ACROSS

Nucleus
MEMBRANES
ER 2.4.1 Thermodynamics of Solute Transport
In the experiment described in Table 2.2, the K concentra-
c
b
tion in maize root sap (which is approximately equal to
a the K concentration of the vacuoles) was 80 times higher
d
Cls than in the external solution. In contrast, the Na concentra-
Medial Golgi tion in the root sap remained lower than that in the external
Trans solution. Such phenomena require both a source of energy
c
and selective transport across the plasma membrane of root
cells.
Transport across plant membranes is facilitated by
transmembrane proteins (Fig. 2.7). These can be clas-
Peroxisome sified into three groups: (i) primary active transporters
Mitochondrion (pumps), in which solute transport is coupled directly
to the hydrolysis of an energy substrate such as ATP or
Chloroplast pyrophosphate (PPi); (ii) secondary active transporters or
PSV ‘coupled transporters’, which harness the electrochemical
gradient of (generally) H to the movement of a solute in
either the same (symport) or opposite (antiport) direction;
Lytic vacuole and (iii) passive transporters, which catalyse the movement
of solutes down their electrochemical gradient. The latter
FIGURE 2.6 Pathways of membrane trafficking. The initial pathways
are divided into (a) endoplasmic reticulum (ER) and Golgi integration
group includes a variety of carriers (uniporters) and chan-
and (b) transport of vesicles between the ER and Golgi. The subsequent nels. Channels can be distinguished from uniport carriers
pathways (c) involve transport of vesicles between the ER and peroxi- by their high catalytic rate, which can exceed 10 million
somes and vacuoles, and between the Golgi and peroxisomes, vacuoles, ions s1 which is several orders of magnitude greater than
chloroplasts, mitochondria and plasma membrane. Feedback signals from uniport carriers (White, 2003). In the next paragraphs, the
the Golgi to the nucleus (d) are thought to regulate aspects of membrane
trafficking. Figure adapted from Matheson et al. (2006).
driving forces for solute movement across membranes are
considered in relation to facilitated diffusion, or ‘passive’
transport, of solutes down their electrochemical gradient
important for the maintenance of membrane functions at by carriers and channels, and to ‘active’ transport of sol-
low temperatures. utes against their electrochemical gradient catalysed by
Cellular membranes are dynamic structures that are pumps and coupled transporters.
continuously remodelled to allow the plant to respond to Under most circumstances, the driving force for the
developmental signals, biotic challenges and environmen- facilitated diffusion of an uncharged solute across a mem-
tal conditions. This remodelling occurs over minutes to brane is its concentration gradient, whereas for an ion it
months, and is supported by complex trafficking pathways is its electrochemical gradient (White, 2003). The Nernst
that deliver lipids and proteins to and from cellular mem- equation (Fig. 2.8) allows the direction of the net dif-
branes (Fig. 2.6). These pathways are functionally linked fusive flux of an ion at a given membrane potential and
through the Golgi apparatus to the endoplasmic reticulum, temperature to be determined. When the cell membrane
plasma membrane, peroxisomes, vacuoles, mitochondria potential is more negative than the Nernst potential, cati-
and chloroplasts (Matheson et al., 2006; Robinson et al., ons can move into the cell, and anions out of the cell, by
2007). The delivery of secretory vesicles to the plasma facilitated diffusion. When the membrane potential is more
membrane can be targeted to specific locations, such as positive than the Nernst potential, the opposite fluxes are
the apex of tip-growing cells, e.g. elongating root hairs favoured. According to the Nernst equation, at 20°C with
or pollen tubes (Cole and Fowler, 2006; Cheung and Wu, a membrane potential of 100 mV, K or Cl would be in
2008; Ishida et al., 2008; Sorek et al., 2009), or to plas- electrochemical equilibrium across the plasma membrane
modesmata (Oparka, 2004; Maule, 2008; Lucas et al., if their concentration in the cytosol were 52 times higher
2009). Thus, membranes are not homogeneous, but pos- (K) or 52 times lower (Cl) than in the external solution
sess domains in which specific lipids and proteins can be (Fig. 2.8). At the same temperature and membrane poten-
clustered, stably or transiently, to improve the efficiency tial, the concentrations of a divalent cation or anion would
of biochemical and physiological processes (Opekarová differ more than 2,700-fold between the cytosol and exter-
et al., 2010). nal medium if it was in electrochemical equilibrium.
GS-X Ca2+ H+ Na+
ATP ATP ATP

H+ K+ H+ NH4+ K+
Symport Antiport Uniport
Pumps Carriers Channel
Primary active transport Secondary Passive transport

active transport
FIGURE 2.7 Nomenclature of transport proteins. Schematic representation of primary active transport mechanisms, such as ABC transporters (e.g.,
glutathione conjugate pump), metal transporters (e.g., Ca2-ATPase) and H-ATPases, secondary active transport mechanisms, such as the K/H sym-
porter or the Na/H antiporter, and passive transport mechanisms, such as the NH4 carrier and the K channel. Figure adapted from White (2003).
(A) environmental signals including exposure to salt and low

EK = RT/zF In ([K]o / [K]i)
temperatures. Under physiological conditions, many cati-
ECl = RT/zF In ([Cl]i / [Cl]o) ons are in electrochemical equilibrium across the plasma
membrane of root cells (White, 2003). However, there
(B) Microelectrode is always a large electrochemical gradient driving Ca2
Cell wall
influx to cells, and, in saline environments, there is also a
large electrochemical gradient driving Na influx. On the
Cytoplasm
other hand, anions cannot be concentrated in the cytoplasm
by facilitated diffusion across the plasma membrane, and
External Vacuole
solution their influx to root cells is often facilitated by symporters
coupled to the proton electrochemical gradient generated
by plasma membrane H-ATPases.
(C) External solution Cytoplasm
(−100 mV)
At the molecular level, facilitated diffusion is mediated
Chemical 1 mM K+ 1 mM K+ by uniporters or channels. Passive transporters facilitat-
equilibrium 1 mM Cl− 1 mM Cl− ing the influx of 10 of the 14 mineral nutrients across the
Electrochemical 1 mM K+ 52 mM K+ plasma membrane of root cells have been reported (Fig.
equilibrium 1 mM Cl− 0.019 mM Cl− 2.9; White, 2003; Gojon et al., 2009; Karley and White,
2009; White and Broadley, 2001, 2003, 2009; Miwa and
FIGURE 2.8 (A) The Nernst equation. The equilibrium potential Fujiwara, 2010; Teakle and Tyerman, 2010). These include
for potassium (EK) and chloride (ECl) are given as a function of R, the
K-channels, such as AtAKT1:AtKC1 of Arabidopsis thal-
gas constant (8.314 V C K1 mol1), T, the absolute temperature, z the
valency of the ion, and their activity concentrations outside (subscript iana, voltage-dependent Ca-channels, cation channels,
o) and inside (subscript i) the membrane. (B) Schematic representation such as those encoded by the cyclic nucleotide gated chan-
of the system used for measuring the membrane potential of plant cells. nel (CNGC) and glutamate receptor (GLR) gene fami-
(C) Example of the calculation of ion distributions at electrochemical lies, ammonium transporters encoded by the ammonium
equilibrium assuming a membrane potential of 100 mV at 20°C.
transporter (AMT) gene family, M, transporters, such as
AtMGT1 and AtMGT10, members of the Zn-regulated
The resting membrane potential of root cells is often transporter (ZRT)-, Fe-regulated transporter (IRT)-like
more negative than 2100 mV (Maathuis and Sanders, protein (ZIP) family, which transport Fe2, Zn2, Cu2
1993; Walker et al., 1996; Britto and Kronzucker, 2006). It and Mn2, Cu transporters encoded by CTR/COPT
is generated primarily by the activity of plasma membrane genes, boric acid channels, formed by nodulin-26-like
H-ATPases encoded by members of the AHA gene fam- intrinsic proteins (NIPs) and plasma membrane intrinsic
ily (Gaxiola et al., 2007). These H pumps are clustered in proteins (PIPs), and, in saline environments, Cl chan-
discrete (micro)domains of the plasma membrane and their nels. However, influx into root cells of nutrients present
activity is regulated by phosphorylation-dependent interac- in the soil solution as anions (e.g., nitrate, phosphate, sul-
tions with cytosolic 14-3-3 proteins in response to diverse phate, molybdate, chloride) is not thought to be mediated
ATP
H+ H+
K+ PPi ATP
NH3
H+ Ca2+
Ca2+ ATP
Pumps
Urea ATP
Ca2+
Cation Amino Zn2+
ATP
acid Heavy ATP
NH4+ Cu2+
K+ metals ATP
Urea
Channels
Ca2+ G-X
B ATP
K+, Ca2+ H+ H+
K+
K+ NO3−
Cation H+
Cl− H+
Na+ H2PO4−
Cl− H+ H+
Anion Ca2+ SO42−, MoO42−
Malate
Anion H+ H+
Organic Mg2+ Cl−
Citrate H+ H+
acid
transporters
Zn2+, Mn2+ K+
Coupled
H+ H+
Cations NA-Fe, NA-Zn
H+ H+
Mg2+ B Amino acid
Carriers
Fe2+, Zn2+ H+ H+
Cu2+, Mn2+ NO3− Cl−
H+ H+
Cu+ SO42− Na+
H+ H+
K+ B
H+ H+
Fe2+, Mn2+ NO3−
Vacuole
Cytoplasm
FIGURE 2.9 Transport proteins of the tonoplast and plasma membrane of plant cells. See text (Section 2.4.1) for details.
by facilitated diffusion but by active transport against their AtSLAC1 protein have been proposed as candidates for the
electrochemical gradient, as discussed below. In the stele, S-type anion channels of root cells (Teakle and Tyerman,
uniporters and channels facilitate the efflux of potassium, 2010).
nitrate, sulphate, phosphate, chloride and organic acids In addition to uniporters and channels, solute transport
from xylem parenchyma cells into xylem vessels in the across membranes can be catalysed by primary or second-
direction of their electrochemical gradients (Section 3.2). ary active transporters that move solutes against their elec-
Similar transport proteins are present in the plasma mem- trochemical gradient. Several ATPases are present in the
branes of other plant cells, where they serve both general plasma membrane of plant cells. These catalyse the efflux
and specific functions. Channels in the plasma membrane of H, Ca2 and heavy metals from the cytoplasm. The
of root cells facilitating the efflux of malate or citrate plasma membrane H-ATPases catalyse H efflux, which
into the rhizosphere, such as members of the Al-activated is then coupled directly, through the proton electrochemi-
malate transporter (ALMT) family and the multidrug and cal gradient, or indirectly, via the cell membrane potential,
toxin extrusion (MATE) protein family, respectively, have to the movement of other solutes. The plasma membrane
been implicated in Al tolerance and improving P availabil- Ca2-ATPases remove Ca2 from the cytosol to maintain
ity in acid soils (Delhaize et al., 2007; Ryan et al., 2011). the low cytosolic Ca2 concentrations required for cell sig-
Channels facilitating the efflux of chloride, such as the nalling (Section 6.6). In the stele, Ca2-ATPases and CPx-
depolarization activated R-type and S-type anion chan- ATPases catalyse the efflux of Ca2 and other divalent
nels (White and Broadley, 2001; Roberts, 2006; Teakle cations from the symplast to the xylem (Section 3.2).
and Tyerman, 2010) in the plasma membrane of root cells, A multitude of secondary active transporters are
may be required for charge compensation of other ion present in the plasma membranes of root cells, which
fluxes. Recently, homologues of the Arabidopsis thaliana couple H influx to the movement of solutes against their
electrochemical gradients (Fig. 2.9). Proton-coupled trans- extremely sensitive to inhibition by Ca2 (Gaxiola et al.,
porters in the plasma membrane of root cells are respon- 2007; Martinoia et al., 2007).
sible for the uptake of anions, such as nitrate (e.g., NRT1 Magnesium is essential for both H-ATPases and H-
and NRT2 transporters), phosphate (e.g., PHT1 transport- PPiases, since their substrates are Mg.ATP and Mg.PPi
ers), sulphate (SULTR1 transporters), chloride and (prob- (White et al., 1990c; Gaxiola et al., 2007). In addition, the
ably) molybdate (White and Broadley, 2001; Buchner H-PPiases require Mg2 for their activity (White et al.,
et al., 2004; Fitzpatrick et al., 2008; White and Hammond, 1990c; Gaxiola et al., 2007). Inorganic pyrophosphate is
2008; Gojon et al., 2009; Miller et al., 2009; Shinmachi generated in several major biosynthetic pathways, such
et al., 2010). In addition, proton/potassium symporters, as starch synthesis or activation of sulphate. Cytosolic
such as those encoded by the KUP/HAK gene family, facil- PPi concentrations generally lie in the range 50–400 µM,
itate K uptake by root cells (White and Karley, 2010), and which is adequate to drive this proton pump (White et al.,
homologues of the maize yellow stripe 1 protein (ZmYS1) 1990c). Under most circumstances, H-PPiases contribute
allow proton-coupled symport of Fe and Zn conjugates far less than H-ATPases to proton transport into the vacu-
(White and Broadley, 2009) into root cells. Proton-coupled ole. Therefore it has been suggested that H-PPiases act as
transporters also alleviate element toxicities by remov- ancillary enzymes to maintain the proton electrochemical
ing chloride, sodium and boron from root cells (White gradient across the tonoplast when the activity of the H-
and Broadley, 2001; Munns and Tester, 2008; Miwa and ATPases is restricted by substrate availability, for example
Fujiwara, 2010). In the stele, proton-coupled transport- during anoxia (White et al., 1990c), or at high tempera-
ers load nitrate and B into the xylem (Miller et al., 2009; tures, which promote protein degradation (Martinoia et al.,
Miwa and Fujiwara, 2010). Similar transport proteins 2007).
are present in the plasma membranes of other plant cells, The proton electrochemical gradient generated by the
where they serve both general and specific functions. The tonoplast H-ATPase and H-PPiase supports the activi-
transport of amino acids, peptides and sugars across the ties of a large number of proton-coupled transporters.
plasma membrane is also catalysed by proton-coupled These catalyse the efflux of K (e.g., NHX and KEA
transporters. transporters), Na (NHX transporters), Ca2 (CAX trans-
The tonoplast of the vacuole similarly contains a porters), NO3 (e.g., AtCLC-a and AtCLC-c), sucrose
variety of primary active transporters, proton-coupled (e.g., AtSUT4), and various divalent cations, including
transporters, uniporters and channels (Fig. 2.9). In cells of Mg2, Zn2 and Mn2 (e.g., CAX, MGT and MTP trans-
higher plants, the electrical potential difference between porters) from the cytosol to the vacuole, and the influx of
the vacuole and the cytosol is about 20 to 60 mV K, nitrate (e.g., AtCLC-a), SO42 (e.g., AtSULTR4-1;
and the pH of the vacuolar sap can be as low as pH 3 AtSULTR4-2) and iron (e.g., AtNRAMP3) from the vacu-
(Martinoia et al., 2007). Based on estimates of solute con- ole to the cytosol in times of high demand for growth
centrations in the cytosol and vacuole, it is thought that (Shigaki and Hirschi, 2006; Martinoia et al., 2007; Gojon
sequestration of K, Na, Ca2, Mg2, Zn2, Mn2 and et al., 2009; Miller et al., 2009; White and Broadley, 2009;
nitrate requires active transport into the vacuole, whereas White and Karley, 2010; Zifarelli and Pusch, 2010). The
the movement of other anions is likely to be passive sequestration of K, Cl and NO3 in vacuoles is impor-
(White and Broadley, 2001, 2003; Martinoia et al., 2007; tant for turgor regulation and the sequestration of Na,
Teakle and Tyerman, 2010; White and Karley, 2010). Ca2 and heavy metals is important to avoid cytoplasmic
The tonoplast contains two distinct types of proton poisoning (Section 17.6). In addition, the sequestration of
pumps, the H-ATPases and the H-PPiases that generate essential elements and metabolites in the vacuole provides
the negative electrical potential across the tonoplast and storage for times of need (Martinoia et al., 2007).
lower the pH of the vacuole (Gaxiola et al., 2007). The The tonoplast also contains Ca2-ATPases (e.g.,
tonoplast H-ATPases of plants are complex oligomeric AtACA4) that pump Ca2 into the vacuole (White and
proteins comprising two subcomplexes: the peripheral V1 Broadley, 2003) and a variety of ATP Binding Cassette
complex, which consists of eight subunits (A, B, C, D, E, (ABC) transporters that protect the cytoplasm by remov-
F, G and H) and is responsible for ATP hydrolysis, and ing heavy metals, oxidation products conjugated to glu-
the trans-membrane V0 complex, which consists of up to tathione and xenobiotics from the cytosol into the vacuole
five subunits (a, c, c, d and e) and is responsible for pro- (Martinoia et al., 2007). These transporters are also
ton translocation (Gaxiola et al., 2007). These subunits are involved in the sequestration of chlorophyll catabolites and
encoded by the VHA genes. Plants possess two distinct natural pigments in the vacuole (Martinoia et al., 2007).
H-PPiases, which are both single subunit enzymes. The Several ion channels have been recorded in the tono-
Type I H-PPiases require K for their activity and are plast. These facilitate the movement of K, Cl, NO3,
relatively insensitive to inhibition by Ca2, whereas Type ammonia, amino acids, urea, Ca2, SO42, HPO42, sugars
II H-PPiases do not require K for their activity and are and organic acids in the direction of their electrochemical
gradients (White and Broadley, 2003; Martinoia et al., 2007;

Teakle and Tyerman, 2010; White and Karley, 2010). The TABLE 2.7 Respiratory energy costs for ion uptake in
rapid efflux of K and Cl from the vacuole, through fast roots of Carex diandra
vacuolar (FV), slow vacuolar (SV) or vacuolar potassium Plant age (days)
(VK) channels and Cl channels, respectively, is required Proportion of total ATP
demand required for 40 60 80
for stomatal closure and other osmotically driven plant
movements (White and Broadley, 2001; White and Karley, Ion uptake 36 17 10
2010). The sequestration and release of NO3, ammo- Growth 39 43 38
nia, amino acids and urea are central to the N economy
Maintenance of biomass 25 40 52
of plants (Martinoia et al., 2007). Aquaporins have been
a
shown to facilitate the transport of ammonia (e.g., AtTIP2;1, Based on Van der Werf et al. (1988).
AtTIP2;3) and urea (e.g., AtTIP1;1, AtTIP1;2; AtTIP2;1,
AtTIP4;1) across the tonoplast (Martinoia et al., 2007;
Miller et al., 2009). The rapid efflux of Ca2 from the vacu- υmax
υ (µmol K+ [g fresh wt]−1 h−1)

ole through SV, voltage-regulated, cADPR-regulated or IP3- 10
regulated channels is important for cell signalling (Section
8.6). The influx of malate through anion channels is a pre-
requisite for crassulacean acid metabolism (CAM), which
separates CO2 fixation from photoassimilation and allows 5
plants to restrict water loss in arid environments by clos-
ing stomata during the day. In CAM plants at night, malate Km
enters the vacuoles as malate2 through an anion channel,
and is accumulated in monovalent and uncharged forms by 0
0 0.05 0.10 0.15 0.20
vacuolar acidification (White and Smith, 1989; Martinoia
K+ concentration (mM)
et al., 2007). Malate is subsequently released to the cyto-
plasm during the day for photoassimilation to occur. FIGURE 2.10 Rate of K uptake (v) as a function of the external con-
From the preceding discussion, it is apparent that pro- centration of KCl () or K2SO4 (Δ); Km  0.023 mM. Adapted from
ton pumps are responsible for energizing solute transport Epstein (1972).
across cell membranes. However, it is important to note
that these pumps not only generate the proton electro- the plasma membrane of root cells occurs when these ions
chemical gradient across the tonoplast and plasma mem- are present at high concentrations in the rhizosphere solu-
brane, and the acidic conditions of the apoplasm (pH ~5.5) tion (Britto and Kronzucker, 2006). As the rhizosphere
and the vacuole (pH 4.5–5.9), but also maintain cytosolic concentration increases, and the rate of unidirectional
pH at its optimal value (pH 7.3–7.6; Felle, 2001). influx increases, the quotient of the unidirectional rates
of ion influx and ion efflux across the plasma membrane
approaches unity (see Fig. 2.13). The energy costs of this
2.4.2 Energy Demand for Solute Transport ‘futile’ cycling represent a substantial proportion of the total
The energy demand for ion uptake by roots can be consid- respiratory energy cost of the root. Similarly, under saline
erable, especially during rapid vegetative growth (Table conditions, a considerable proportion (almost all) of the
2.7). Early calculations suggested that, in seedlings, up total energy budget of the root is expended on the removal
to 36% of the total respiratory energy cost, expressed as of sodium (Britto and Kronzucker, 2006, 2009) and chlo-
ATP consumption, is required for ion uptake; this propor- ride (Teakle and Tyerman, 2010) from the symplasm.
tion declines in older plants in favour of ATP demand for
growth and maintenance of biomass (Table 2.7). Kurimoto 2.4.3 The Kinetics of Solute Transport in
et al. (2004) subsequently calculated that up to 76% of
total respiratory energy cost was required to support low-
Plant Roots
affinity nitrate influx across the plasma membrane of As a general rule, the rate of solute uptake by plant cells,
cereal roots. Although several of the assumptions of these excised plant tissues and roots of intact plants saturates
calculations have been criticized, recent studies addressing with increasing external solute concentration. Emanuel
the weaknesses of the original calculations also found very Epstein and colleagues in the early 1950s suggested that
high energy costs of ion uptake (Britto and Kronzucker, this relationship was similar to that between an enzyme
2006, 2009; Teakle and Tyerman, 2010). and its substrate (Fig. 2.10). In their analogy, the transport
Recently, it has been observed that high rates of appar- protein was an enzyme that catalysed the movement of its
ently ‘futile’ cycling of K, NH4, NO3 and SO42 across substrate from one side of a membrane to the other. Using
the terms from enzymology, they defined the relationship marked effect on the uptake rate. These observations led to
between the rate of transport of a solute and its concentra- the hypothesis of dual systems for K transport, with System
tion by the Michaelis-Menten equation: I having a higher selectivity than System II.
In view of the usually very low concentrations, par-
V (Vmax S) /( K m S) ticularly of P and K, in soil solutions (Section 13.2) and
results of ion uptake studies in the low concentration range
where V is the rate of solute transport at a solute concen- (10 µM), the term Cmin was introduced to define the con-
tration of S, Vmax is the maximal rate of solute transport, centration at which net uptake of ions ceases before the
and Km is the Michaelis constant, which is the solute ions are completely depleted (Fig. 2.11). The Cmin con-
concentration at which half the maximal transport rate is centration is an important factor in ion uptake from soils,
reached. The Km value reflects the affinity of the trans- because it is the lowest concentration at which roots can
porter for the solute; just as in enzymatic reactions it indi- extract an ion from the soil solution. Cmin concentrations
cates the affinity of the enzyme for its substrate. differ considerably between plant species (Asher, 1978).
When assayed at low concentrations in the exter- For P, for example, a value of 0.12 µM has been found
nal medium, solute uptake is often described well by this in tomato (Itoh and Barber, 1983a), 0.04 µM in soybean
equation, as illustrated by K uptake by barley roots (Fig. (Silberbush and Barber, 1984) and 0.01 µM in ryegrass
2.10). It is evident from this experiment that the relation- (Breeze et al., 1984). For K, the corresponding values
ship between K uptake and K concentration in the exter- were 2 µM in maize (Barber, 1979) and 1 µM in barley
nal medium is the same whether the source of K is KCl (Drew et al., 1984). Cmin concentrations for nitrate can
or K2SO4. However, as we shall see later, when substrate vary from between more than 50 µM to less than 1 µM
concentrations are higher, the accompanying anion can depending not only on the plant species but also on the
have a significant effect on the uptake rate of a cation and
vice versa. As a first approximation, in the low concentra-
tion range, Michaelis-Menten kinetics can also be applied TABLE 2.8 Boron concentration in shoots and shoot
to describe uptake rates of many other solutes including dry weight of two barley genotypes with increasing B
the anions nitrate, phosphate, sulphate and chloride (e.g., supply
Epstein, 1972; Deane-Drummond, 1987; Siddiqi et al.,
1990; Teo et al., 1992; Laine et al., 1993; Wang et al., B supply (µM)
1993; White and Broadley, 2001; Li et al., 2007), the cati- 0 2.5 7.5 15
ons ammonium, calcium, magnesium, manganese, zinc B concentration (mg kg1 dw)
and cadmium (e.g., Kelly and Barber, 1991; Huang et al.,
1992a; Kronzucker et al., 1998; Rawat et al., 1999; Sadana Schooner 5.6 10.0 22.1 46.4
et al., 2005; Broadley et al., 2007; Lux et al., 2011) and Sahara 3771 2.5 5.5 7.8 11.7
chelates such as Fe-phytosiderophores (von Wirén et al., Shoot dry weight mg per plant
1995). A low-capacity saturable uptake system can some-
Schooner 129 140 132 121
times be discerned in B-deficient plants (e.g., Dannel
et al., 2000); but the relationship between the uptake of Sahara 3771 74 84 92 107
B and its concentration in the external solution is often Based on Nable et al. (1990b).
reported to be linear in B-replete cells (Seresinhe and
Oertli, 1991) and plants (Table 2.8). However, the relation-
ship between solute concentration and its uptake by roots
cannot always be fitted to a simple Michaelis-Menten Imax
equation. This is a consequence both of the theoretical
I (uptake rate)
limitations of Michaelis-Menten kinetics and the presence

of multiple mechanisms for the transport of a particular Imax(Cs-Cmin)
I=
solute (White, 2003). Km + (Cs-Cmin)
The original concept of a single protein-mediated mech-
anism of ion transport (one carrier system for each ion) did Km
not sufficiently describe the kinetics of uptake when wide
concentration ranges were tested. At concentrations above External concentration (CS)
Cmin
1 mM, for example, the kinetics of K uptake differ consid-
erably from those at lower concentrations (Epstein et al., FIGURE 2.11 Schematic presentation of the relationships between
1963). The apparent selectivity of transport is lower (Na uptake rates (net influx  I) of ions and their external concentrations;
competes with K) and the accompanying anion has a Cmin  net uptake zero (influx  efflux).
environmental conditions (Deane-Drummond and Chaffey,

1985; Marschner et al., 1991). For ammonium, Cmin con-
centrations decreased from 30 to 1.5 µM as the root zone
temperature increased (Marschner et al., 1991).
External For the kinetics of ion uptake by plants, the Michaelis-
solution
φCV φVC φCX
Menten equation has been modified to include the param-
eter Cmin, and the term I, designating unidirectional influx,
JCV Xylem has replaced the term V (Fig. 2.11). Very often, though,
JOC only the net uptake of ions is determined experimen-
Vacuole tally, which is the net result of influx and efflux across the
φCO
φOC plasma membrane (Fig. 2.12). Efflux can become similar
in magnitude to influx, particularly at extreme low or high
Cytoplasm external concentrations, and therefore can be an important
component in determining net uptake (Fig. 2.13; Elliott
et al., 1984; Britto and Kronzucker, 2006). It is also note-
worthy that, at a given external concentration, the efflux of
FIGURE 2.12 Nomenclature of unidirectional (ø) and net (J) solute a particular mineral nutrient can be many times higher from
fluxes across the plasma membrane between cytoplasm (c) and the exter- roots of plants sufficiently supplied than from roots of defi-
nal solution (o) or xylem (x), and across the tonoplast between the cyto- cient plants (e.g., McPharlin and Bieleski, 1989; Lee et al.,
plasm (c) and the vacuole (v) of a stereotypical root cell. Figure adapted
from White and Broadley (2001).
1990; Topa and Sisak, 1997).
1.0 1.0
0.8 0.8
0.6 0.6
K+
0.4 0.4 NO3−
0.2 0.2
0 0
0.1 1 10 100 0.1 1 100
0.8 0.8
Efflux:Influx ratio
0.6 0.6
0.4 0.4 SO42−
0.2 NH4+ 0.2
0 0
0.001 0.1 10 0.01 1 100
1.0 1.0
0.8 0.8
0.6 0.6
0.4 0.4
Na+ Cl−
0.2 0.2
0 0
0.1 10 100 0.1 1 100
External ion concentration
FIGURE 2.13 Ratios of efflux to influx across the plasma membrane for K, NO3, NH4, SO42, Na and Cl. Data from various studies, each
study plotted as a different symbol. Adapted from Britto and Kronzucker (2006).
The efflux of ions and other solutes is affected by sev- uptake in this experiment, therefore the contribution of
eral factors: (i) the integrity of the plasma membrane, (ii) increased efflux at higher root P concentrations cannot
the presence of transport proteins allowing efflux, (iii) the be evaluated. However, for nitrate, ammonium, potas-
electrochemical driving force for transport, and (iv) the sium, phosphate and sulphate, there is evidence that both
concentration of the solute in the cytoplasm. In pea, for increased efflux and reduced influx contribute to the
example, the initial high rate of net uptake of sulphate by decline in net uptake when internal concentrations are
S-deficient roots placed in a solution containing sulphate increased (Lee et al., 1990; Britto and Kronzucker, 2006).
decreases to about 30% within one hour due to a marked In the high concentration range (1 mM), a linear rela-
increase in sulphate efflux, despite a slight increase in influx tionship is often found between external concentrations and
(Deane-Drummond, 1987; Bell et al., 1995). Similarly, for the rate of ion uptake by plant roots. This has been observed,
nitrate and ammonium, the efflux component can account for example, for the anions nitrate, phosphate, sulphate
for a high proportion – almost 40–50% of the influx – and chloride (Loneragan and Asher, 1967; Epstein, 1972;
most probably due to the high concentrations of nitrate Borstlap, 1983; Clarkson and Saker, 1989; Siddiqi et al.,
and ammonium in the cytoplasm (Britto and Kronzucker, 1990; White and Broadley, 2001; Li et al., 2007), the cati-
2003, 2006). The rapid exchange between ions in the exter- ons ammonium, potassium, sodium, calcium, magnesium,
nal solution and in the cytoplasm is reflected in low half- iron and zinc (Epstein, 1972; Borstlap, 1983; Wang et al.,
times for exchange (t{1/2}), which are between 7–14 min for 1993; Rawat et al., 1999; White, 2001; Vallejo et al., 2005;
ammonium (Kronzucker et al., 1998), 10–50 min for potas- Broadley et al., 2007), for boron (Dannel et al., 2000), and
sium (White et al., 1991; Szczerba et al., 2006), 7–75 min for Fe-phytosiderophores (von Wirén et al., 1995). Several
for calcium (White et al., 1992), 10–20 min for sulphate explanations for the linear relationship (formerly defined
(Deane-Drummond, 1987; Bell et al., 1995), 10–20 min as System II; Epstein, 1972) have been proposed. The first
for chloride (Britto et al., 2004), 4–107 min for nitrate explanation is that it reflects influx through non-saturating
(Lee and Clarkson, 1986; Macklon et al., 1990; Britto and transport proteins, perhaps ion channels, in the plasma
Kronzucker, 2003) and 23–115 min for phosphate (Lee membrane of root cells. The second explanation is that it is
et al., 1990; Macklon et al., 1996). These rates of exchange the consequence of rapid chelation or metabolism of a sol-
with the cytoplasmic pool are usually orders of magnitude ute in the cytoplasm, or its removal by sequestration in the
faster than the rates of exchange with the vacuole (e.g., vacuole or transfer to the xylem, which maintains the elec-
Macklon et al., 1990; White et al., 1991, 1992; Bell et al., trochemical gradient, and reduces efflux, across the plasma
1995). membrane. The third explanation is that it represents a non-
The parameters of ion uptake kinetics are also strongly saturating, apoplasmic flux to the xylem. However, given the
affected by the nutritional status of plants. Roots of plants usually low ion concentrations in soil solutions, the ecologi-
deficient in a particular nutrient generally exhibit a greater cal significance of the low-capacity, non-saturating mecha-
Imax and a lower Cmin for that nutrient than plants suffi- nisms for the nutrition of plants grown in natural soils has
ciently supplied. Occasionally, but not always, deficient been questioned. There are, however, at least two excep-
plants also exhibit a lower Km. An example is given in tions: plants growing in saline soils (Section 17.6), and the
Table 2.9 for P. In plants with greater tissue P concentra- uptake of mineral nutrients from the apoplast following
tions, Imax for P uptake is substantially lower, and Km their long-distance transport in the xylem (Section 3.2) and
is also slightly lower. The Imax values were based on net phloem (Section 3.3).
TABLE 2.9 Short-term P uptake parameters of soybean plants with different P

nutritional status
P concentration
(mg kg1 dw)
Plants grown at P Imax (mol cm1
concentration (µM) Shoot Root sec1  1014) Km (µM)
0.03 2.2 2.3 17.6 1.6
0.3 3.4 3.0 16.9 1.7
3.0 5.9 5.6 6.5 1.2
30.0 6.6 9.0 3.7 1.0
Based on Jungk et al. (1990).
2.5 FACTORS AFFECTING ION UPTAKE The volume of root tissue accessible for apoplasmic
BY ROOTS solute movement, the free space, represents only a small
fraction of the total root volume. For example, the free
2.5.1 Influx to the Apoplasm space is estimated to occupy 5% of a maize root (Shone
Before reaching the plasma membrane of root cells, ions and Flood, 1985). The presence of this free space ena-
must pass through the cell wall. In general, neither diffu- bles individual cortex cells to contribute to solute uptake
sion nor mass flow of ions or other low-molecular-weight from the external solution. Solute concentrations in the
solutes is restricted at the external surface of the roots. The free space depend on various factors such as the capacity
cell walls and water-filled intercellular spaces of the root for solute uptake by epidermal cells, the presence of root
cortex are, to a certain extent, accessible to these solutes hairs, the solute concentration in the rhizosphere solution
from the external solution. and the rate of transpiration. As shown more than 50 years
The main barrier to solute flux through the apoplasm of ago by Vakhmistrov (1967), at low external concentrations
young roots is the endodermis, the innermost layer of cells root hair formation is usually extensive and the uptake of
of the cortex (Fig. 2.14). Suberization of the radial and mineral nutrients is limited mainly to the rhizodermal cell
transverse walls of the endodermal cells (the Casparian layer, i.e. the outer-most cells of the cortex. This is particu-
band) creates an effective barrier against solute move- larly relevant for roots growing in soil, where the impor-
ment into the stele. In most angiosperm species suberiza- tance of root hairs for the acquisition of nutrients present
tion of the radial and transverse cell wall is also found in at low concentrations in the soil solution or with restricted
the hypodermis, or exodermis (cell layer below rhizoder- soil mobility, such as P, has been clearly demonstrated
mis; Enstone and Peterson, 1992; Ma and Peterson, 2003). (Gahoonia and Nielsen, 2004; Gahoonia et al., 2006; Zhu
These barriers may also protect the inner cortex from colo- et al., 2010).
nization by microorganisms, for example preventing the
colonization of sorghum roots by endophyte Polymyxa spp. 2.5.2 Effects of pH
(Galamay et al., 1992).
The pH of the external solution can have profound effects
Suberization of the exodermis generally occurs after
on the uptake of nutrients by plant roots. These can be
the formation of the endodermal Casparian band, par-
divided into three broad categories: (i) effects of solution
ticularly in fast growing roots (Ma and Peterson, 2003).
pH on the chemical species present in solution, (ii) effects
There are different views on the effectiveness of the exo-
of apoplasmic pH on the concentrations of ions present in
dermis as a barrier to solute movement through the root
the apoplasm, and (iii) influence of rhizosphere pH for the
apoplasm (Clarkson et al., 1987; Enstone and Peterson,
proton electrochemical gradient and the driving force for
1992). However, since the development of the exodermis
proton-coupled solute transport. In addition, solution pH
generally occurs after that of the endodermis, its function
can affect ion transport by protonation/deprotonation of
is thought to be largely structural. In plants adapted to sub-
amino-acid residues of transport proteins.
merged conditions, the exodermis serves another function,
The pH of the soil solution influences the availability
namely as an effective barrier against oxygen diffusion
of cations and anions for root uptake (White and Broadley,
(leakage) from the root aerenchyma into the rhizosphere
2009). In alkaline soils, the availability of P, Zn, Fe, Mn,
(e.g., Soukup et al., 2007).
Cu and B is very low, whereas in acid soils, plant growth is
mainly limited by toxic concentrations of Al3 and Mn2
in the rhizosphere. In addition, the pH of the external solu-
CB = Casparian band
O = Xylem tion also determines the chemical species present in the
rhizosphere. This is particularly relevant to the uptake of
Rhizodermis solutes that can be protonated and are transported across
the plasma membrane as specific chemical species, such
Cortex
as boron, phosphate and ammonium. The rate of B uptake
Endodermis CB Root hairs
decreases strongly when the pH of the external solution is
increased (Fig. 2.15). This pattern is closely related to the
Stele
decrease in the ratio of boric acid, which is the substrate of
the transporter catalysing boron uptake by root cells (Miwa
and Fujiwara, 2010), to the borate anion. Similarly, the rate
Exodermis
of phosphate uptake decreases as the pH of the external
solution increases (Fig. 2.16). This can be explained by a
FIGURE 2.14 Schematic representation of cross section of a differenti- decrease in the concentration of H2PO4, the substrate of
ated root zone of maize. the proton-coupled phosphate symporter in the plasma
100 −Ca2+
µmol K+ [(g fresh wt)−1 3 h−1]

Relative B uptake (%)
+20
80
60 +10 +Ca2+
40
0
20
−10
0
6 7 8 9 10 11
2 3 4 5 6 7 8 9 10
Solution pH pH
FIGURE 2.15 Relative uptake of B by barley roots as a function of the
FIGURE 2.17 Net uptake of K by barley roots from solutions con-
external solution pH. Uptake at pH 6  100 at each supply concentration.
taining 5 mM KBr as a function of the pH of an external solution with
Solid line: percentage of undissociated H3BO3. Key for B concentrations
(Ca2) or without calcium (Ca2). Modified from Jacobson et al.
(mg l1): 1.0 (open triangle), 2.5 (open square), 5.0 (open circle), 7.5 (filled
(1960).
triangle), 10.0 (filled square). Based on Oertli and Grgurevic (1975).
the rate and selectivity of ion influx to root cells. As the

external pH is lowered, the effective CEC of the apoplasm
100 decreases, particularly in monocotyledonous species due to
binding of H to the cation exchange sites. Thus, apoplas-
80
Relative values (%)
mic pH can affect ion uptake.

The pH of the rhizosphere solution can also affect ion
60 uptake by altering both substrate (H) concentration and
the electrochemical driving force for proton-coupled sol-
40
ute transport. A decrease in pH can increase the activity
of proton-coupled solute transporters and enhance anion
20
uptake. Thus, the uptake of anions is generally either not
affected or stimulated by low pH. In short-term experi-
0
4 5 6 7 8 9 ments with maize roots, decreasing the external pH from
Solution pH 8 to 4 increased nitrate influx by a factor of about 10
(McClure et al., 1990b) and phosphate uptake by a factor
FIGURE 2.16 Relationships between solution pH and the proportion of of about 3 (Sentenac and Grignon, 1985). For phosphate,
H2PO4 (dashed line) in solution, and the uptake of phosphate (solid line)
and sulphate (dotted line) by bean plants. Data are expressed as relative
this increase at low pH was also observed when the con-
values. Figure adapted from Hendrix (1967). centration of the monovalent species (H2PO4) was kept
constant. In contrast, the efficiency of H efflux decreases
as the external solution becomes more acidic and, conse-
membrane, in the pH range of 5.6 to 8.5 (Fig. 2.16). In quently, the membrane potential of root cells decreases
contrast, there is a smaller effect on the uptake rate of from about 150 mV at pH 6 to 100 mV at pH 4
sulphate, since in this pH range only the divalent anion (Dunlop and Bowling, 1978). Accordingly, the driving
SO42 occurs. The effects of the pH of the external solu- force for cation uptake is reduced. In general, the uptake of
tion on ammonium uptake by plant roots are more com- cations, such as K, is inhibited by low pH of the external
plex. At high external pH, ammonium uptake increases solution, although Ca2 often has an ameliorating effect
sharply, probably due to an increase in the proportion of (e.g., Fig. 2.17).
the uncharged species NH3 and NH4OH. The contrasting effects of external pH on cation and
In relation to the effects of apoplasmic pH on the anion uptake are well-documented phenomena, for exam-
uptake of solutes by root cells, it has been noted previously ple, in rice (Zsoldos and Haunold, 1982) and soybean
that both cell walls and biological membranes contain (Rufty et al., 1982b). In the latter case, a decrease in the
charged groups and that ions interact with these groups. pH of the external solution from 6.1 to 5.1 resulted in an
Generally, the strength of these interactions increases with increase in the ratio of anion to cation uptake from about
valency. Fixed negative charges can influence both the 1.0 to 1.25. In long-term growth experiments, the contrast-
absolute and relative concentrations of cations in the apo- ing effects of external pH on the uptake of cations and
plasm and, thereby, ion movements in the apoplasm and anions are reflected in the nutrient composition of plants
140
120
100
(%) 80
60
40
20
4.0
3.3
5.5
8.5
0
K Ca Mg Mn N(NO3−) P S
FIGURE 2.18 Nutrient concentrations (expressed on a dry weight basis and as a percentage of the concentration observed in plants grown at pH
8.5) in shoots of bean (Phaseolus vulgaris) grown in solutions with pH 8.5, 5.5, 4.0 and 3.3, respectively, as indicated in the columns for K. Data
recalculated from Islam et al. (1980).
(Fig. 2.18), with tissue cation concentrations being more

affected than tissue anion concentrations. TABLE 2.10 K and P uptake by barley
The effect of external pH on N uptake depends on plants at different oxygen partial pressure
whether N is supplied as ammonium (NH4) or nitrate around roots
(NO3). As is to be expected, lowering the external pH Relative uptakea
increases the uptake of NO3, but decreases the uptake of Oxygen partial
pressure (%) K P
NH4 (Zsoldos and Haunold, 1982). Interpreting results
with both forms of N at different external pH are, however, 20 100 100
complicated by side-effects, such as changes in cation– 5 75 56
anion balance in the plants and on root metabolism and
0.5 37 30
function.
Based on Hopkins et al. (1950).
2.5.3 Metabolic Activity

The accumulation of ions and other solutes against a con-
centration gradient requires an expenditure of energy,
either directly or indirectly. The main source of energy in TABLE 2.11 Sugar concentration, respiration (O2
non-photosynthesizing cells and tissues (including roots) uptake), and N uptake in barley roots after root
is respiration. Thus, all factors that affect respiration can excision
influence ion accumulation. Net uptake
Oxygen. As oxygen tension is lowered, the uptake (µmol g1 dw min1)
Time (h) Sugar
of ions such as potassium and phosphate is decreased,
after excision (µmol g1 dw) O2 NH4 NO3
particularly at very low oxygen tensions (Table 2.10).
Consequently, oxygen deficiency is one of the factors that 0 82 4.5 1.8 1.5
can restrict plant growth in poorly aerated substrates (e.g., 3 51 3.3 1.1 1.0
waterlogged soils; Section 17.4). Recalculated from Bloom and Caldwell (1988).
Carbohydrates. The main energy substrates for res-
piration are carbohydrates. Therefore, in roots and other
non-photosynthesizing tissues, under conditions of lim-
ited carbohydrate supply from a source (e.g., leaves), a
close correlation can often be found between carbohydrate relationships are of particular ecological importance, for
concentration and the uptake of ions. For example, carbo- example, when leaves are removed (grazing, cutting) or
hydrate concentration, respiration and N uptake decrease in dense plant stands when light supply to the basal leaves
within a few hours after excising roots which cuts off the is limited, since the basal leaves are the main source of
photosynthate supply from the shoot (Table 2.11). These carbohydrates for the roots.
Distinct diurnal patterns in solute uptake (maxima dur- Recently, it has been suggested that diurnal fluctua-
ing the day, minima during the night) have been observed tions in the uptake of nitrate, phosphate, sulphate, ammo-
for nitrate, phosphate, ammonium, potassium, iron and nium, potassium and iron may be related to the delivery of
zinc (e.g., Clement et al., 1978b; Zhang et al., 1991b; Le sucrose to the root in the phloem through the regulation of
Bot and Kirkby, 1992; Macduff et al., 1997; Cesco et al., the expression of genes encoding proteins catalysing their
2002; Vert et al., 2003; Louahlia et al., 2008). Root carbo- transport across the plasma membranes of root cells (Lejay
hydrate concentration may act as a coarse control for ion et al., 2003; Vert et al., 2003; Hammond and White, 2008;
uptake and is one of the factors responsible for the diur- Liu et al., 2009; Vance, 2010).
nal fluctuations in ion uptake. However, in maize roots, for There is evidence that the delivery of sucrose via the
example, diurnal fluctuations in nitrate uptake were only phloem can act as a systemic signal informing the root of
loosely related to root carbohydrate content (Fig. 2.19). In the shoot N and P status (Hermans et al., 2006; Hammond
soybean growing under short-day conditions, the typical and White, 2008) and that the increase in uptake capac-
diurnal fluctuations of nitrate uptake could be reversed by ity in roots of plants lacking sufficient N or P for maxi-
an intervening 3 h period of low light (i.e., imitating long- mal growth (Section 2.5.6) is effected by sucrose-induced
day conditions, repressed flower initiation); uptake rates of expression of nitrate and phosphate transporters.
nitrate were then twice as high during the night as com- Temperature. Physical processes such as exchange
pared with the day (Raper et al., 1991). adsorption of cations in the AFS are only slightly affected
by temperature (Q10  1.1–1.2, with Q10 referring to the
change in a reaction or process imposed by a change in
6 NO3− 0.6
temperature by 10°C). However, chemical and biochemical
Water soluble carbohydrates
reactions show much greater temperature dependence with

Q10  2 and Q10  2, respectively. The Q10 value for the
(% in dry matter)
uptake of ions from solutions of low concentration often

Nitrate reductase activity
[µmol NO2 9 g fw−1 h−1]
4 0.4
exceeds 2, at least within the physiological temperature
range (e.g., Fig. 2.20; Clarkson et al., 1988; Wang et al.,
1993). Ion uptake is more temperature dependent than respi-
2 0.2 ration, especially at temperatures below 10°C. Furthermore,
at very high temperatures root respiration further increases
whereas ion uptake declines (Fig. 2.20), indicating that
membrane transport and respiration are not coupled directly.
0 0 In studies of temperature effects on ion uptake, two
0 6 12 18 24
phenomena are often studied: (i) the immediate effects of
Hours
an abrupt change in root temperature, which occur within
FIGURE 2.19 Diurnal fluctuations in nitrate uptake (solid line), nitrate seconds and reflect the direct effects of temperature on the
reductase activity and concentration of water-soluble carbohydrates in uptake system, and (ii) the long-term effects of growing
maize roots. Nitrate uptake: relative values, uptake at end of the light plants at a particular root temperature. The latter effects
period  100. Adapted from Keltjens and Nijenstein (1987).
are manifest after several days or weeks of growth at a
1.4
40
1.2
[µmol (g fw)−1 h−1]
1.0
30
Ion uptake
[µmol (g fw)−1 h−1]
0.8
O2 consumption
0.6 20
0.4
10
0.2
0 0
0 10 20 30 40
Temperature (C)
FIGURE 2.20 Rates of respiration () and uptake of P () and K () from solutions containing 0.25 mM K and 0.25 mM P by maize root segments
at different temperatures. Adapted from Bravo and Uribe (1981).
particular root temperature and include adaptive responses, than ammonium in cold-sensitive plants like cucumber
for example changes in root membrane properties. The (Tachibana, 1987), and in cold-tolerant species, such as
latter effects are of greater ecological significance. Such barley and ryegrass, the strong preference for ammonium
studies often compare plants exposed to different root tem- compared with nitrate uptake is little affected by the tem-
peratures, but the same shoot temperature. This creates a perature of the root zone (Macduff and Jackson, 1991;
temperature differential between the root and the shoot, Clarkson et al., 1992). Compared with Ca and Mg, uptake
and it is noteworthy that the shoot meristem of, for exam- rates of K are often more affected by root zone tempera-
ple, gramminoid plants is close to stem base. Thus, differ- tures. In winter wheat, the increase in K/(Ca  Mg) ratios
ent temperatures in the rooting medium will influence cell in the shoots with increasing root zone temperature may
division and cell elongation in the shoot. In the long term, cause tetany in grazing beef cattle on winter wheat forage
such experimental systems can affect the growth rates of (Miyasaka and Grunes, 1990).
root and shoot quite differently and, thus, the root/shoot In contrast to plants grown in solution culture, the
biomass ratio (Clarkson et al., 1988, 1992). Accordingly, roots of plants grown in soil must forage for many immo-
long-term effects of root temperature on ion uptake can bile nutrients (Engels and Marschner, 1990; Rengel,
include feedback regulation via plant demand, an example 2001; Lynch, 2007; White and Hammond, 2008; White
of which is shown for maize in Table 2.12. and Broadley, 2009). In soil-grown plants, therefore, root
In maize, low root temperatures (12°C) decrease shoot temperature can affect the uptake of nutrients additionally
and root growth and uptake rates of nitrate and potassium, through effects on root growth rate and root system mor-
as might be expected for a cold-sensitive plant species. phology (Section 13.3).
However, the reduction in ion uptake at low tempera-
ture was not a temperature effect on the roots per se, but
2.5.4 Interactions between Ions in the
reflected feedback regulation via lower shoot demand.
This was shown by increasing the temperature of the shoot Rhizosphere
growing zone (stem base) (24/12°C). Shoot growth was In the preceding sections, for the sake of simplicity, the
strongly increased (i.e., the demand for nutrients) and so transport of a particular ion was treated as a singular
were the uptake rates of nitrate and potassium per unit root process. In reality, however, the transporters catalysing ion
weight (Table 2.12). Similarly, in other graminaceous spe- uptake are rarely specific and ions can compete directly
cies, poor growth at low root temperatures is generally not for transport. This competition is influenced by the proper-
caused by limited uptake of nutrients such as N, K or P ties of the transporter itself and by the concentrations of
(Clarkson et al., 1986, 1992). different ions in solution. Solutes that are not transported
Low root temperatures can affect the uptake of nutri- can also interact with transport proteins altering their activ-
ents differently, P uptake usually being reduced more than ity. In addition, there may be indirect interactions between
the uptake of other nutrients (e.g., Engels and Marschner, ions as a result of their transport across the plasma mem-
1992a; Engels, 1993). The uptake rate of nitrate seems to brane, for example via effects on membrane potential
be more strongly reduced at low root zone temperatures through the movement of charge, or via effects on the pro-
ton electrochemical gradient through the coupling of solute
transport to proton movements.
TABLE 2.12 Shoot and root growth and uptake of
nitrate and K by maize plants grown at different root 2.5.4.1 Competition
zone temperatures (RZT) and the temperatures at the Transport proteins catalyse the movement of nutrients
stem base (shoot growing zone temperature, SGT) for from the rhizosphere solution to the cytoplasm across the
eight days plasma membrane of root cells (Fig. 2.9). Competition
Temperature treatment between ions of the same valency for entry to a chan-
(SGT°C/RZT°C) nel protein or for binding to a carrier protein is common,
24/24 12/12 24/12 whether these ions are ultimately transported or merely
inhibit the transport process. Such competition occurs
Shoot growth (g fw day ) 1 1.91 0.32 1.34
particularly between ions with similar physicochemi-
Root growth (g fw day1) 0.85 0.20 0.26 cal properties (valency and ion diameter), for example
Nitrate uptake (pmol g1 fw day1) 6.40 4.20 7.60 between the alkali cations potassium (K), rubidium
(Rb), cesium (Cs) and sodium (Na), or between the
K uptake (pmol g1 fw day1) 2.50 1.20 3.10
Group II divalent cations calcium (Ca2), strontium (Sr2)
After Engels and Marschner (1992a). and barium (Ba2). It is important to note, however, that
the inhibition of transport of a particular ion by another ion
TABLE 2.13 Interactions between the uptake of NH4 TABLE 2.14 Uptake of labelled Mg2 (28 Mg) by barley
and K by maize rootsa seedlings without or with supply of K and Ca2
(0.25 mM each)
Concentration in roots (µmol g1 fw)
Ammonium Potassium Mg2 Uptake (µmol Mg2 (10 g)1 fw 8 h1)
(NH4)2SO4 (mM) K K K K MgCl2 

MgCl2 MgCl2  CaSO4 CaSO4  KCl
0.00 6.9 6.7 8.2 53.7
Roots 165 115 15.0
0.15 7.3 7.1 6.7 48.4
Shoots 88 25 6.5
0.50 17.1 13.5 8.9 41.1
Based on Schimansky (1981).
5.00 29.4 31.5 9.3 27.1
Based on Rufty et al. (1982a).
a
Duration of the experiment: 8 h;  K indicates addition of 0.15 mM K;
calcium concentration constant at 0.15 mM.
TABLE 2.15 Uptake rates of Mn and Mg by roots of
soybean plants at increasing Mn concentrations in the
does not necessarily imply that the inhibitory ion is itself substrate
transported. Manganese supply (µM)
Radioactive rubidium (86Rb) has often been used as a
Nutrient (μmol g 1
root dw) 1.8 90 275
tracer to study K transport in plants, although this can
give misleading results under certain circumstances (Behl Mn 0.5 3.1 4.8
and Jeschke, 1982). In general, transport proteins catalys- Mg 121.8 81.1 20.2
ing K transport across the plasma membrane of root cells, Based on Heenan and Campbell (1981).
such as K-channels, cation-channels, and proton-coupled
K symporters, do not differentiate between K and
Rb for transport (White, 1997; Maathuis and Amtmann,
1999; Vallejo et al., 2005; Pyo et al., 2010; White and ammonium by root cells will be determined not only by
Karley, 2010). However, the major K-channel in roots of competition of NH4 for cation transporters, but also by
Arabidopsis thaliana (AtAKT1:AtKC1) is relatively imper- rhizosphere pH and cytosolic tolerance of NH4 and NH3
meable to Cs, which inhibits K influx through this uptake.
channel (White and Broadley, 2000). By contrast, proton- Applications of K and Ca fertilizers often induce Mg
coupled K symporters, such as AtHAK5, can transport deficiency in crop plants. This is partly a consequence of
both K and Cs into root cells and, in plants sufficiently cations, such as K and Ca2, inhibiting Mg2 uptake
supplied with K, the influx of both Cs and Na is thought by plant roots (Table 2.14). The presence of Mn2 in the
to occur largely through non-selective cation-channels rhizosphere also inhibits Mg2 uptake by roots (Table
(White and Broadley, 2000; Qi et al., 2008; Munns and 2.15), but has little effect on the uptake of K (Heenan and
Tester, 2008). Campbell, 1981). This presumably reflects the contrasting
The competition between potassium (K) and ammo- specificity of the transporters responsible for the uptake of
nium (NH4) is difficult to explain simply by competi- each cation.
tion for a single transport process at the plasma membrane Competition also occurs between anions for uptake by
(Table 2.13). Whereas NH4 is quite effective in inhibiting root cells. Some well-known examples are competition
K influx, the reverse (inhibition of NH4 uptake by K) between sulphate and molybdate, sulphate and selenate,
is rarely observed (e.g., Mengel et al., 1976; Rufty et al., selenite and phosphate, and phosphate and arsenate.
1982a; Shaviv et al., 1987). This may be explained by two Sulphate and molybdate are thought to enter root cells
phenomena. First, it has been reported recently that, in through the same proton-coupled symporters (Fitzpatrick
Arabidopsis thaliana, NH4 competes with K for trans- et al., 2008). Increasing the sulphate concentration in
port through both AtAKT1 and AtHAK5 (ten Hoopen the rooting medium reduces molybdate uptake strongly.
et al., 2010) and also reduces the expression of AtHAK5 Hence, S fertilization may be an effective tool to reduce
(Qi et al., 2008), whereas K does not appear to affect excessive Mo uptake thereby improving plant growth
the expression or activity of the major NH4 transporter, and animal nutrition in soils containing high concentra-
AtAMT1. Second, a substantial proportion of ammonium tions of Mo (Pasricha et al., 1977; Chatterjee et al., 1992).
may not be taken up as NH4 through transporters such as However, the competition may become a critical factor in
AtAMT1 but as NH3 (Section 2.5.2). Thus, the uptake of soils containing little Mo.
The interactions between selenate and sulphate are also

quite distinct, and of considerable practical importance in TABLE 2.16 Chloride concentrations in roots
view of the absolute requirement for selenium of humans and shoots of barley plants at different nitrate
and animals and the detrimental effects of excessive sele- concentrations in the nutrient solution
nium in the diet (White and Broadley, 2005a, 2009). Concentation in Chloride content
Sulphate and selenate are taken up into root cells through nutrient solution (mM) (µmol g1 fw)
the same proton-coupled symporters (White et al., 2007b).
Cl NO3 Roots Shoot
Increasing sulphate concentration in the substrate strongly
decreases selenate uptake by roots and selenium accumu- 1 0 52 93
lation by plants, suggesting direct competition between 1 0.2 26 73
selenate and sulphate for transport (White et al., 2007b).
1 1.0 13 54
On the other hand, increasing selenate concentration in the
substrate often increases sulphate uptake and accumulation 1 5.0 9 46
by plants (White et al., 2004, 2007b; Stroud et al., 2010), Based on Glass and Siddiqi (1985).
possibly by interfering with the regulation of expression
or activity of high-affinity sulphate transporters by plant
S-status (White et al., 2004, 2007b).
Antagonistic interactions between selenite and phos- This reduction seems to be the result of negative feedback
phate, and also between phosphate and arsenate, are from nitrate stored in the vacuoles of root cells (Glass and
thought to occur because these anions are transported by Siddiqi, 1985). Similarly, nitrate uptake is reduced when
the same proton-coupled symporters into root cells (Li roots contain high chloride concentrations, and chlo-
et al., 2008c; Zhao et al., 2010). In Holcus lanatus L., ride accumulated in the vacuoles seems to be particularly
arsenate-tolerant and non-tolerant genotypes exist, and effective in this respect (Cram, 1973). The active efflux of
arsenate uptake is much lower in the tolerant genotypes chloride and nitrate from the cytoplasm into the vacuole is
(Meharg and Macnair, 1992). The low arsenate uptake rate catalysed, in part, by the same proton-coupled transporters
is achieved by suppression of the P deficiency-induced (members of the CLC family) and several anion channels
high affinity uptake system in the tolerant plants. Similar also facilitate the movement of both Cl and NO3 across
mechanisms of arsenic tolerance have been observed in the tonoplast (White and Broadley, 2001; Martinoia et al.,
other plant species. For example, mutants of Arabidopsis 2007; Teakle and Tyerman, 2010; Zifarelli and Pusch,
thaliana with defective phosphate transport are more tol- 2010). Thus, it is possible that the two anions compete for
erant to arsenate (Shin et al., 2004). Arsenite and undis- transport across the tonoplast, which affects their accumu-
sociated methylated arsenic species are taken up by roots lation in vacuoles, cytoplasmic concentrations and uptake.
through the silicon transport pathway via members of the In addition, several anion channels and proton-coupled
nodulin 26-like intrinsic protein (NIP) family (Zhao et al., symporters in the plasma membranes of root cells appear
2010). Members of this family also transport a range of to facilitate the transport of both chloride and nitrate
small neutral molecules including ammonia, urea, boric (White and Broadley, 2001; Roberts, 2006), suggesting
acid and silicic acid (Maurel et al., 2008; Wallace et al., further interactions in the pathways of their uptake and
2006; Miwa and Fujiwara, 2010). accumulation.
The inability of transport proteins to differentiate effec- Interactions between nitrate and chloride during their
tively between K and Rb, Ca2 and Ba2, SO42 and uptake and accumulation in vacuoles are of great impor-
SeO42, and phosphate and arsenate illustrates that the tance for crop production. The competing effect of chlo-
selectivity of transport proteins in the plasma membrane of ride can be used to decrease the nitrate content of plant
root cells does not indicate any essential role for an ele- species such as spinach which tend to accumulate large
ment in the plant, but merely reflects the physicochemi- amounts of nitrate for use as an osmoticum. On the other
cal similarities between ions. Plant roots may be unable hand, in saline soils, the competing effect of chloride on
to exclude many non-essential or toxic ions from the root nitrate uptake may impair N uptake by the plants (Bernal
symplasm. This has important practical implications, et al., 1974). Under these conditions increasing nitrate sup-
for example, for the entry of heavy metals into the food ply can be an effective means to improve the N nutritional
chain via their uptake and accumulation by plants (e.g., status of the plants and simultaneously prevent chloride
Marschner, 1983). toxicity in sensitive plant species (Section 17.6).
Another distinct type of anion competition occurs An interesting case of the indirect regulation of trans-
between chloride and nitrate. Chloride concentrations port by nutrients is the inhibition of nitrate uptake, and
in plant tissues, particularly in roots, can be reduced stimulation of chloride uptake, by ammonium supply
strongly by increasing nitrate availability (Table 2.16). (Lee and Drew, 1989; Xu et al., 2000; Miller and Cramer,
2004). In almost all cases, increasing the availability of to the resealing of the plasma membrane following dam-
ammonium strongly suppresses nitrate uptake. By contrast, age (Schapire et al., 2009). These functions of Ca2 are
increasing nitrate supply generally has little or no effect on reflected, for example, in the higher rates of efflux of low-
ammonium uptake (Breteler and Siegerist, 1984). Thus, molecular-weight solutes across the plasma membrane of
when nitrogen is supplied as NH4NO3, ammonium is taken Ca-deficient cells when faced with environmental chal-
up in preference to nitrate. In Norway spruce, the rhizo- lenges, such as low temperatures or mechanical damage.
sphere ammonium concentration must fall below about Calcium can be removed fairly readily from its binding
100 µM NH4 before nitrate uptake occurs (Marschner sites at the outer surface of the plasma membrane, for
et al., 1991). In short-term experiments with barley, exter- example by chelators (Van Steveninck, 1965), or can be
nal ammonium inhibited net influx of nitrate within 3 min, exchanged by high concentrations of H or metal cations
and upon removing ammonium from the external solution including Na (Lynch et al., 1987), which will increase
net influx of nitrate resumed within 3 min (Lee and Drew, solute efflux.
1989). Such immediate effects suggest that they arise from Rhizosphere Ca2 concentration also influences the
the effect of ammonium on the electrochemical gradients selectivity of ion uptake, and the relative accumulation
supporting nitrate uptake across the plasma membrane. of K and Na in particular. For example, in the absence
of Ca2 there are clear differences in the K/Na uptake
2.5.4.2 Effects of Extracellular Calcium ratio between the ‘natrophobic’ maize and the ‘nat-
rophilic’ sugar beet. However, the presence of Ca2 in
An example of synergism, first discovered by Viets (1944), the rhizosphere solution shifts the uptake ratio in favour
is the stimulation of cation and anion uptake by extracel- of K at the expense of Na in both species (Table 2.18).
lular Ca2 at low rhizosphere pH (Table 2.17; Fig. 2.17). These shifts in K/Na uptake ratio are likely to be due
It is thought that this phenomenon is the result of Ca2 to the fact that extracellular Ca2 inhibits Na influx
counteracting the negative effects of high H concentra- through voltage-insensitive cation-channels (White, 1999;
tions on plasma membrane integrity or the activity of the Maathuis and Amtmann, 1999; Munns and Tester, 2008),
plasma membrane H-ATPase. Calcium, as a divalent but has little effect on K influx through inward-rectifying
cation, stabilizes membranes through interactions with K-channels (White, 1997a; Maathuis and Amtmann,
the negatively charged headgroups of phospholipids and, 1999). High Ca2 concentrations in the soil solution are
thereby, influences membrane function. It also contributes particularly beneficial for the maintenance of high K/Na
uptake ratios in saline environments as they increase plant
salt tolerance.
TABLE 2.17 K and Cl uptake in barley roots with or 2.5.4.3 Cation–Anion Relationships
without Ca2 supply with external pH 5.0
The uptake of cations and anions occurs through differ-
Uptake rate (µmol g1 dw (2 h) 1) ent transport proteins (Fig. 2.9), therefore direct inter-
External K net Cl net actions between cations and anions for uptake are rare.
solution (mM) K influx uptake Cl influx uptake However, the uptake of one nutrient can influence the
uptake of another indirectly through effects on the mem-
0.1 KCl 116  3 117  6 35  1 34  4
brane potential, the proton electrochemical gradient or via
0.1 KCl  137  2 140  7 53  3 52  4 feedback regulation through plant growth or metabolism.
1.0 CaSO4 The stimulation of cation uptake by anions, and of anion
uptake by cations, is observed frequently, and is generally
TABLE 2.18 K/Na selectivity of roots with or without Ca2 supply

Uptake rate (µmol g1 fw (4 h) 1)
Maize Sugar beet
External solution NaCl 
KCl (10 mM each) Na K Na  K Na K Na  K
Ca 9.0 11.0 20.0 18.8 8.3 27.1
1 mM CaCl2 5.9 15.0 20.9 15.4 10.7 26.1

a consequence of the necessity to maintain charge balance. solutions. Different uptake rates of cations and anions
However, synergism in ion uptake can also be the result of require both compensation of electrical charges and regu-
a general increase in root metabolic activity when nutrients lation of cellular pH. At high external concentrations these
are supplied after a period of deprivation. requirements become a limiting factor for the uptake of K
When present at low concentrations in the rhizosphere, when accompanied by SO42 and for Cl when accompa-
the rate of uptake of a cation is not affected by the accom- nied by Ca2 (Table 2.19).
panying anion and vice versa, as shown in Table 2.19 for Different rates of cation and anion uptake by roots can
K and Cl. At high external concentrations, however, cause perturbations of intracellular pH. The stabilization
an accompanying ion that is taken up relatively slowly of cytosolic pH in the range of 7.3 to 7.6 is achieved by
can reduce the uptake of an oppositely charged ion that is the so-called cellular pH-stat, which consists of two com-
transported at a faster rate: for example, SO42 depresses ponents: the biophysical pH stat, characterized by proton
K uptake and Ca2 depresses Cl uptake from single-salt transport across the plasma membrane or tonoplast (Fig.
2.21), and the biochemical pH stat, which involves produc-
tion and consumption of protons through metabolism and
is achieved by the formation and removal of carboxylic
TABLE 2.19 Rate of K and Cl uptake by maize plants groups (Britto and Kronzucker, 2005; Miller and Cramer,
with different accompanying Ions 2004; Peuke, 2010). The functioning of the biochemi-
Uptake rate (µmol g1 fw h1) cal pH stat is thought to be reflected in the net changes
in organic acid concentrations in roots in response to an
K from Cl from
Concentration imbalance in cation–anion uptake ratio (Table 2.20). When
(mM) KCl K2SO4 KCl CaCl2 K2SO4 is supplied, the excess cation uptake is compen-
0.2 1.6 1.6 0.8 0.7 sated for by an equivalent synthesis of organic acid ani-
ons and when CaCl2 is supplied the excess anion uptake is
2.0 2.7 1.9 2.0 1.0
compensated for by an equivalent decrease in the synthesis
20.0 5.7 2.2 4.3 2.1 of organic acid anions. These changes in organic acid con-
Recalculated from Lüttge and Laties (1966). centrations are also reflected in the rates of CO2 fixation in
the roots (dark fixation).
External
solution Plasma Cytoplasm
(Apoplasm) Vacuole
membrane Tonoplast
CO2
A Cat+ Cat+
(K+)
(K+) R COO
H+ R COO−
H+ H+
R COO−
An−
An−
(SO42−) (SO42−)
B Cat+ Cat+
(Ca2+)
(Ca2+)
CO2
H+ H+ H+
An− An−
2−) (NO32−)
(NO3
R COO− + H+ R COO− + H+
FIGURE 2.21 Model for internal pH stabilization and for charge compensation at different ratios of cation:anion uptake from the external solution. A.
Excessive uptake of cations (Cat), for example, with K2SO4 supply. B. Excessive uptake of anions (An), for example, with Ca(NO3)2 supply.
The main reactions involved in the traditional concept the net H efflux was 2.15 µmol g1 root fresh weight
of the biochemical pH stat in relation to different cat- h1, leading to a decrease in the pH of the external solu-
ion–anion uptake ratios are shown schematically in Fig. tion from 5.60 to 5.12 (Hiatt and Hendricks, 1967). The
2.21. Excessive cation uptake (A) results in an increase cation–anion balance in plants and the consequences for
in cytosolic pH, which increases the synthesis of organic rhizosphere pH and mineral nutrition of plants has been
acids. This produces anions (R.COO) for pH stabiliza- reviewed by Haynes (1990) and, in the context of nitrogen
tion and charge compensation and enables the subsequent nutrition, by Britto and Kronzucker (2005).
transport of cations and anions either into the vacuole or In the cytoplasm, the equilibrium between carboxyla-
the shoot. By contrast, excessive anion uptake (B) is cor- tion (CO2 fixation) and decarboxylation is thought to be
related with a decrease in cytosolic pH, which stimulates regulated by the pH sensitivity of two enzymes, phospho-
the decarboxylation of organic acids from the storage pool enolpyruvate (PEP) carboxylase and malic enzyme (Fig.
(i.e., the vacuoles). This causes an increase in pH as decar- 2.22). An increase in pH activates PEP carboxylase (reac-
boxylation consumes protons. In addition to increases or tion (1)), and both the rate of CO2 fixation and the syn-
decreases in root concentrations of organic acid anions, thesis of oxaloacetate are increased. After oxaloacetate is
the biochemical pH stat also affects the pH in the root reduced to malate by the enzyme malate dehydrogenase,
apoplasm and external solution with excess cation uptake the malate can be directly transported into the vacuoles
increasing proton efflux thus decreasing the external pH (reaction (2)), where it acts as a counterion for excess cati-
whereas excess anion uptake increases it. In the experi- ons (Fig. 2.21A). Alternatively, malate can be incorporated
ment reported in Table 2.20 when K2SO4 was supplied, into the cytoplasmic pool of the organic acids of the Krebs
cycle, and another organic acid from this pool (e.g., citric
acid) can be transported into the vacuole. An oxalate-based
biochemical pH stat may play an important role in plant
TABLE 2.20 Relationship between the uptake of cations species that accumulate large amounts of oxalate, such as
and anions and the organic acid concentration of members of the Chenopodiaceae (Davies, 1986). When
isolated barley roots anions are taken up in excess (Fig. 2.21B), the pH of the
cytoplasm decreases and malic enzyme (reaction (4)) is
Uptake
External (µmol g1 fw) Change in 14
CO2 activated, leading to the decarboxylation of malate and the
solution organic acid fixation production of CO2. As a result of these reactions, the cyto-
(mM) Cations Anions (µmol g1 fw) (relative) plasmic pH is stabilized and the cation–anion ratio in the
2 K2SO4 17 1 15.1 145 cells maintained. This biochemical pH stat responds rap-
1 KCl 28 29 0.2 100 idly to supply of K2SO4, PEP carboxylase activity being
increased by 70% within 20 min (Chang and Roberts,
1 CaCl2 1 15 9.7 60 1992).
Based on Hiatt (1967a, b) and Hiatt and Hendricks (1967). Nitrogen nutrition (NH4; NO3; N2 fixation) has
a strong effect on cation–anion relationships in plants
CO2
COOH
COOH 1
Phosphoenol C O Oxaloacetate
pyruvate C O P PEP Carboxylase
CH2
(PEP)
CH2 COOH
Malate NADPH+H+
ADP
dehydrogenase NADP+
ATP
COOH
COOH
Malic enzyme HC OH Malate
C O
4 CH 2
CH3 CO2
Pyruvate COOH
Krebs
cycle
3 Vacuole
FIGURE 2.22 Model of the pathways of CO2 fixation (‘dark fixation’) and decarboxylation. Reactions (1)–(4) are explained in the text.
because about 70% of the cations and anions taken up by as observed for NO3-fed plants. However, under condi-
plants are either NH4 or NO3 (Van Beusichem et al., tions where NO3 uptake and assimilation are impaired
1988). Nitrogen nutrition affects both organic acid metabo- and cation/anion uptake ratio is high, a strong decrease
lism and the element composition of plant tissues (Table in the pH of the external solution has been observed.
2.21). Plants supplied NH4 are generally characterized by This phenomenon occurs in many plant species with, for
a high cation/anion uptake ratio and plants supplied NO3 example, P deficiency (Schjorring, 1986), Zn deficiency
by a low cation/anion uptake ratio. However, the effects of (Cakmak and Marschner, 1990) and Fe deficiency in dicot-
NH4 and NO3 on organic acid metabolism differ from yledonous plants.
those anticipated from Fig. 2.21, since N assimilation in In plants supplied NH4, net proton efflux and mainte-
roots is correlated with the production or consumption of nance of the cellular pH stat becomes increasingly difficult
protons. The shoots of higher plants have a limited capac- in roots when the pH of the external solution is low. The
ity to dispose of protons, thus, NH4 assimilation takes presence of high NH4 concentrations in the rhizosphere
place in roots (Engels and Marschner, 1993). The assimila- causes a reduction in both the cytoplasmic and vacuolar
tion of NH4 produces protons, thus, despite a high total pH of root cells (Gerendas et al., 1990). Poor growth of
cation uptake by NH4-fed plants, the pH in the cytoplasm NH4-fed plants at low external pH is probably related
decreases during NH4 assimilation and must be stabilized to the difficulty in maintaining cytosolic pH homeostasis
both by enhanced proton excretion and the decarboxyla- in the face of high NH4 fluxes across the plasma mem-
tion of organic acids (Fig. 2.21B). The protons are effluxed brane of root cells, together with cation–anion imbalance,
to the external solution in equimolar amounts to the NH4 and the high energy costs incurred by the futile cycling
taken up (Marschner et al., 1991) or to the excess cat- of ammonium across the plasma membrane (Miller and
ion uptake (van Beusichem et al., 1988). In contrast, the Cramer, 2004; Britto and Kronzucker, 2006).
assimilation of NO3 is correlated with an approximately Maintenance of cytoplasmic pH homeostasis involves
equimolar consumption of H (Raven, 1986). Depending costs in terms of energy, photosynthate and water (Raven,
on whether NO3 reduction and assimilation take place 1985). This is particularly true in relation to N nutrition.
in the root or the shoot, carboxylates are either produced When both NH4 and NO3 are supplied, cytoplasmic pH
in the roots or transported in the phloem from the shoots homeostasis may be achieved by similar rates of H pro-
to the roots to maintain charge balance (Peuke, 2010). duction (NH4 assimilation) and H consumption (NO3
Legumes dependent on biological nitrogen (N2) fixation assimilation) and thus have a very low energy requirement
are characterized by a cation/anion uptake ratio 1 and (Raven, 1985; Allen et al., 1988). This may explain, in
have higher tissue concentrations of organic acid anions part, why optimal growth for many plant species is usually
and greater proton efflux than plants supplied NH4 (Allen obtained with a mixed supply of NH4 and NO3.
et al., 1988).
The pH of the external solution is strongly influenced 2.5.5 External Concentration
by the form of plant nitrogen nutrition due to differences
in cation/anion uptake ratio, nitrogen assimilation and cel- The relationship between the rate of influx (I), or uptake,
lular pH stabilization (Fig. 2.23). When plants with pref- of an ion and its concentration in solution (S) can usually
erential NO3 reduction in the roots, such as sorghum, are be described by Michaelis-Menten kinetics in the low con-
supplied NO3, the external pH usually increases consid- centration range: the flux saturates, transport appears to be
erably with time. When they are supplied NH4NO3, after selective and is closely coupled to metabolism. In contrast,
preferential uptake of NH4 and depletion of external at high external ion concentrations, the uptake rate is often
NH4, a transient decrease in the pH of the external solu- linearly related to solute concentration (I  kS) through
tion during NH4 uptake is followed by an increase in pH, a proportionality parameter (k), is not very selective, and
TABLE 2.21 Ionic balance in shoots of castor oil plants grown with different forms of N supply
Cations Anions
2 2 2
Form of N supply K 
Ca Mg Total NO3 
H2PO4 
SO4 Cl Organic acidsa Total
NO3  99 85 28 212 44 18 11 2 137 212
NH4  55 43 22 120 0 23 33 5 59 120

Van Beusichem et al. (1988).
a
Calculated from the difference of cations–anions.
8
NO3- TABLE 2.22 Influx of nitrate into barley roots without
7
and with an induced high capacity nitrate uptake system
6 NO3 Influx (µmol g1 fw h1)
pH
5 NO3- + NH4+ External conc. Induced Induced

(mM NO3) Non-induced 1 daya 4 daysa
4
0.02 0.10 2.75 1.54
NH4+
3 0.30 0.39 5.27 2.86
20.0 11.54 20.87 8.02
0 1 2 3 4 5 6 7
Time (days) Based on Glass et al. (1990).
a
Pretreatment with nitrate for one or four days.
FIGURE 2.23 Time course of changes in the pH of the external solu-
tion in sorghum plants supplied with 300 mg l1 total N as only NO3,
only NH4, or both at a ratio of 8 NO3 to 1 NH4 as their N source.
Redrawn from Clark (1982b). The kinetic parameters for a given nutrient are influ-
enced greatly by plant nutritional status and nutrient
availability in the rhizosphere. In general, deficiency of a
K+ (P; NO3−; SO4−) nutrient leads to an increase in the capacity of the root sys-
tem to take up that nutrient (Siddiqi et al., 1990; Rawat et
Net uptake rate
al., 1999; Buchner et al., 2004; Li et al., 2007; White and

Na+ (Mg; Ca)
Broadley, 2009; White and Hammond, 2010; White and
Karley, 2010). For example, in plants grown in the absence
of nitrate, (non-induced) roots possess only a low capac-
B ity (constitutive) influx system for nitrate (Table 2.22).
This constitutive nitrate influx system does not saturate
as the external nitrate concentration is increased, suggest-
External concentration
ing that the transport protein has a low affinity for nitrate.
FIGURE 2.24 Schematic representation of the relationships between However, within 20 min of supplying nitrate, a high-affin-
the rates of uptake of K, Na and B by barley roots and the concentrations ity, high-capacity nitrate influx system is induced (Table
of KCl, NaCl and B in the external solution. The relationships between 2.22). Four days after induction, the capacity for nitrate
the uptake rates of other nutrients and their concentrations in the external
influx is lower, suggesting negative feedback regulation of
solution are indicated in brackets.
the activity of this transport system.
As a rule, optimal growth can be achieved at concentra-
tions in the range of the high-affinity system for nutrients
is not particularly sensitive to temperature or metabolic such as K, P and N when these are supplied continuously
inhibitors. The typical relationship between external con- to plants in the external solution (Asher and Edwards,
centration and uptake rate of K is presented in Fig. 2.24 1983). Similarly, for the micronutrient cations Zn, Cu,
schematically, without consideration of Cmin. Similar Mn and Fe, the concentration of the free metal species
uptake isotherms have been obtained for many nutrients, (Zn2, Cu2, Mn2, Fe3) in the external solution required
although their kinetic parameters (Km, Imax, Cmin, k) differ. for optimal growth derived from chelate-buffered nutri-
The concentrations in soil solutions are usually low for ent solutions, suggest that nutritional requirements can be
K (1 mM) and ammonium (100 µM), and extremely met by extremely low concentrations of free cations in the
low for phosphate (10 µM), boron (10 µM), nickel external solution; in the order of 109 to 1012 for Mn2,
(10 µM), zinc (1 µM), copper (1 µM), molybdate Zn2 and Cu2, and perhaps even lower for Fe2/3 pro-
(1 µM), manganese (1 µM), and iron (0.1 pM in cal- vided a continuous supply is maintained (Bell et al., 1991;
careous and alkaline soils). On the other hand, the concen- Laurie et al., 1991; Webb et al., 1993; Fox et al., 1996;
trations of Ca2, Mg2 chloride, sulphate and nitrate are Degryse et al., 2006).
often in the millimolar range, especially in arable soils Thus, under optimal conditions in which a constant
(Table 13.3; Tyler and Olsson, 2001; White and Broadley, nutrient supply is maintained (e.g., in nutrient solutions),
2001, 2009). Therefore acquisition of most nutrients will only very low concentrations of nutrients are required in
require significant energy input, and a selective, high- the external solution for maximal plant growth. At higher
affinity transport system to enable the plant to satisfy plant supply, higher uptake rates reflect what is known as ‘luxury
demand and avoid ion toxicities. uptake’. In soil-grown plants in general, and under field
conditions in particular, the conditions in the root environ-

ment are far from optimal and the maintenance of a con- TABLE 2.23 Relationship between tissue
stant nutrient supply to the root surface is unlikely to occur. concentration and K influx to barley roots
Higher external concentrations and luxury uptake in pre- K Concentration K Influx (µmol
ceding periods can be important in providing an internal (µmol g1 fw) g1 fw h1)
reserve for periods of high demand or interrupted root sup-
20.9 3.05
ply. This also holds true for natural vegetation experiencing
transient nutrient flushes under favourable weather condi- 32.1 2.72
tions (Rorison, 1987; Millard and Grelet, 2010). 47.9 2.16
In both soil and nutrient solutions, essential ele-
57.8 1.61
ments can be supplied in concentrations so high that they
From Glass and Dunlop (1979).
become toxic to plants; e.g. Mn in acid soils, B in sodic
(Na-rich) soils, and Mn and Fe in waterlogged or flooded
soils (White and Brown, 2010). However, there is consid-
erable genotypic variation in the uptake of nutrients, and
these differences, both between and within plant species, 4 100
Root NO3− concentration (mol g−1)

have important consequences for ecology and agriculture
NO3− influx [nmol g−1 h−1]

(White and Brown, 2010). The ability to grow well in soils 3 75
with high concentrations of elements allows plants to sur-
vive in these soils and also enables the development of 2 50
plants for the phytoremediation of contaminated land. For
example, in barley, the cultivar Sahara 3771 has a lower B
uptake capacity than the cultivar Schooner, and therefore 1 25
requires a higher concentration in the external solution for
optimal growth (Table 2.8). This can be a disadvantage 0
in low B soils, but is an effective mechanism in avoid- 0 24 48 72 96 120 144
Duration of NO3− pretreatment (h)
ing B toxicity when plants are grown in soils with high B
availability.
FIGURE 2.25 Nitrate concentration () and nitrate influx () to roots
The ability to grow well in soils with low availability of barley plants with different nitrate (NO3)-pretreatment. Adapted from
of nutrients confers an ecological advantage in these envi- Siddiqi et al. (1989).
ronments. Genotypic variation within crop species in the
nutrient supply required for optimal growth is well docu-
mented, for example for N, P and K (Hirel et al., 2007;
Rengel and Damon, 2008; White and Hammond, 2008;
Fageria, 2009). Such genotypic variation can be used to that, as the tissue concentration of a particular mineral
improve fertilizer use efficiency in agriculture through the element increases, its influx declines, and vice versa.
development of crops that acquire and/or utilize mineral An example of this feedback regulation is shown for K
elements more effectively. influx in barley roots in Table 2.23. Similar relationships
between internal concentrations and influx rate are well
documented, for example, for nitrate (Fig. 2.25), P (Table
2.5.6 Plant Nutritional Status 2.9), sulphate (Fig. 2.26), Fe, Zn and Cu (Broadley et al.,
The rate of uptake of a nutrient at a given external concen- 2007; White and Broadley, 2009). Immediate effects on
tration is often determined by plant growth rate which is nutrient influx are due to post-translation modifications
thought to affect the uptake of a particular mineral nutri- of regulatory components or transport proteins, whereas
ent through plant nutritional status (e.g., Clement et al., longer-term effects are mediated by transcriptional
1978a; Clarkson et al., 1988; Laine et al., 1993; Walker responses. The mechanisms that may be involved in this
et al., 2001). Nutrient uptake responds rapidly to fluctua- feedback regulation are summarized in Fig. 2.27.
tions in root nutrient concentrations and more slowly to The uptake of NH4 and NO3 is closely related to the
long-term changes in plant demand or external nutrient N status of plants. For example, NH4 uptake capacity is
supply. A rapid decrease in the net uptake of a nutrient by negatively correlated with the concentrations of NH4 and
roots upon an abrupt increase in its external concentration certain amino acids, such as glutamine and asparagine, in
can be the consequence of an increase in its cytosolic con- the roots (Causin and Barneix, 1993; Rawat et al., 1999;
centration and increased efflux across the plasma mem- von Wirén et al., 2000). Accordingly, NH4 uptake capac-
brane (Britto and Kronzucker, 2006). It is also observed ity increases rapidly within a few days after the withdrawal
1400 Shoot
(A) (B)
[nmol (g root fresh wt)−1 h−1]
1200
Plasma 8 Tonoplast
1000 membrane
SO42−
800
Influx
P 7
600 i
400 Gene
P expression
SO4 2−
i
200
Protein
0 synthesis
0 1 2 3 4 5 0 10 20
Time (hours) Protein
Time (days)
modification
FIGURE 2.26 Time course of changes in the influx of sulphate (SO42)
and phosphate (Pi) in roots of barley plants deprived of external sulphate 1 3
supply for up to 5 days (A) and then resupplied with sulphate for up to 24
hours (B). Adapted from Clarkson and Saker (1989).
2 4
5 6
of N supply (Lee and Rudge, 1986; Rawat et al.,

9
1999). In Arabidopsis thaliana, the increase in NH4
uptake capacity is correlated with a decrease in glutamine
Apoplasm Cytosol Vacuole
and increased expression of the AtAMT1 gene (Rawat et al.,
1999). A decrease in NH4 efflux is also observed, but this FIGURE 2.27 Model for the regulation of nutrient uptake by roots via
is not the major factor responsible for increasing net NH4 plant nutritional status. Changes in gene expression, protein synthesis and
uptake in N-starved plants (Morgan and Jackson, 1988). protein modification in response to either the cytosolic concentration of
The regulation of NO3 uptake involves the induction of the mineral nutrient or its metabolites can regulate: (1) nutrient influx via
the number and activity of transporters, and the electrochemical gradient
a high-capacity, high-affinity uptake system and the nega- across the plasma membrane; (2) nutrient efflux via nutrient concentra-
tive feedback regulation of NO3 uptake by increasing tion in the cytosol, the number and activity of transporters, and the elec-
internal NO3 concentrations (Table 2.22). In non-induced trochemical gradient across the plasma membrane; (3) nutrient influx
plants, NO3 supply rapidly increases both NO3 influx and to the vacuole via nutrient concentration in the cytosol, the number and
NO3 concentrations in root tissues. Later, the increase in activity of transporters, and the trans-tonoplast electrochemical gradi-
ent; (4) nutrient efflux from the vacuole via nutrient concentration in the
NO3 concentrations reduces NO3 influx. This negative vacuole, the number and activity of transporters, and the trans-tonoplast
feedback regulation may be caused by high NO3 con- electrochemical gradient; (5) complexation in the cytosol, sequestration
centrations in the vacuoles and follow a turgor-regulated in organelles and metabolism of nutrients, which affect the concentrations
event (Glass, 1983). However, negative feedback regula- of the nutrient and its metabolites in the cytosol; (6) complexation or pre-
tion by elevated concentrations of reduced N in the form cipitation of the nutrient in the vacuole, which affects its ability to exit
the vacuole; (7) transport of a nutrient or metabolite from the root to the
of the amino acids glutamine and asparagine (Lee et al., shoot in the xylem, which affects their cytosolic concentrations; (8) trans-
1992; Louahlia et al., 2008) or of NH4 is a more likely port of a nutrient or metabolite from the shoot to the root in the phloem,
explanation, which is consistent with the inhibitory effects which affects their cytosolic concentrations; (9) modification of the rhizo-
of NH4 supply on NO3 uptake. It is also noteworthy that sphere through the secretion of protons, enzymes and organic solutes to
increased root sucrose concentrations appear to increase increase the concentration of the mineral nutrient in the apoplasm, and its
availability for influx.
the uptake of both ammonium and nitrate through up-
regulation of genes encoding high-affinity transport pro-
teins (Lejay et al., 2003; Louahlia et al., 2008; Girin et al., status. Induction of the transport system for sulphate
2010). requires transcription and protein synthesis, as does the
In a similar manner, sulphate uptake is regulated by induction of enzymes involved in the S assimilatory path-
plant S status. The dynamics of this feedback regulation way (Hawkesford and De Kok, 2006; Hell et al., 2010).
on the uptake of sulphate is shown in Fig. 2.26. Without Early studies suggested that sulphate stored in the vacuoles
external sulphate supply, the capacity of barley roots for played an important role in negative feedback regulation of
sulphate uptake (influx) increases rapidly within 3–5 days, sulphate uptake (Cram, 1983). However, it is now thought
but decreases strongly within a few hours of sulphate that the accumulation of reduced S compounds, such as
resupply and is lost within one day of sulphate resup- cysteine or reduced glutathione (GSH), are the dominant
ply. The influx rate of P is unaffected by the S nutritional signals of tissue S status that regulate sulphate uptake
Although the transport systems induced by nutri-

TABLE 2.24 Phosphorus concentrations in tissues of ent deficiencies are generally specific for the nutrient
barley plants following the supply of P to plants grown the plant lacks, they can also transport other elements.
without P For example, the uptake of selenate increases in sulphur-
P concentration (µmol P g1 dw)a deficient barley plants, and the uptake rate of arsenate
increases in P-deficient barley plants (Lee, 1982). This is
7 days P 7 days P likely to be a consequence of an increased abundance of
8 days Pb 1 day Pc 3 days Pd
the HvSULTR1 proton-coupled sulphate symporter and
Shoot total 49 (20) 151 (61) 412 (176) of the HvPht1;1 and HvPht1;2 proton-coupled phosphate
Youngest leaves 26 (5) 684 (141) 1647 (493) symporters, respectively (Smith et al., 1997; Buchner
et al., 2004; Schunmann et al., 2004; Christophersen et al.,
Roots 43 (24) 86 (48) 169 (94)
2009). Similarly, caesium uptake increases in K-deficient
Based on Clarkson and Scattergood (1982). Arabidopsis thaliana as a consequence of the up-regulation
a
Numerals in parentheses are relative values; 100 represents control with
continuous phosphorus supply of 150 µM P throughout the experiment. of the AtHAK5 gene, which encodes a high-affinity plasma
b
Eight days of growth without P. membrane proton-coupled K (and Cs) symporter (Qi
c
Seven days of growth without P and 1 day of growth upon addition of P
(150 µM). et al., 2008; White and Karley, 2010). Unusual and unex-
d
Seven days of growth without P and 3 days of growth upon addition of pected responses also occur sometimes. In tomato roots,
P (10 µM).
for example, Cd uptake rate increases as the Cd concen-
tration in the root increases (Petit et al., 1978). This may
reflect the induction of synthesis of compounds, such as
metallothioneins or phytochelatins, which chelate and
and assimilation (Hawkesford and De Kok, 2006; Hell detoxify heavy metals. A similar mechanism may be
et al., 2010). These compounds can be the product of involved in the differences in the rate of Cu uptake in
root cell metabolism or, in the case of GSH and its pre- Cu-sufficient and Cu-deficient plants: on resupplying Cu
cursor γ-glutamylcysteine, can be translocated from the to deficient plants the uptake rate is much lower than in
shoots to the roots as a systemic signal of shoot S status Cu-sufficient plants (Jarvis and Robson, 1982).
(Herschbach and Rennenberg, 2001; Hell et al., 2010). The As observed in the preceding paragraphs, the relation-
expression of genes involved in S uptake and assimilation ships between the rate of uptake of a particular nutrient
are also controlled by the delivery of sucrose (Lejay et al., and plant nutritional status cannot always be explained
2003) and a regulatory microRNA (miRNA395; Pant et al., satisfactorily by consideration of root tissue concentration
2008, 2009; Kragler, 2010; Liang et al., 2010) from the alone; feedback regulation of ion uptake by shoot nutri-
shoot. tional status is also evident (Fig. 2.27). Such feedback con-
Similarly to the examples for nitrate and sulphate, P trol is essential for the coordination of nutrient uptake by
uptake (influx) capacity increases after P is withheld from the demand of the plants for growth. Variation in the sup-
the external solution (White and Hammond, 2008). This ply of sugars to the roots (Hermans et al., 2006; Hammond
is correlated with increased transcription of genes encod- and White, 2008; Peuke, 2010), or general differences in
ing proton-coupled P transporters. This transcriptional the rates of root export of nutrients in the xylem to the
response appears to be mediated through the interplay of shoot, can be considered as coarse feedback mechanisms.
biochemical signals indicating root and shoot P status. It For example, an increased supply of sucrose in the phloem
is thought that low root P status initiates a complex regu- leads to greater root biomass and up-regulates the expres-
latory cascade through the PHR1 transcription factor and sion of genes encoding transporters for nitrate, phosphate,
that the increased transport of sucrose and microRNA sulphate, ammonium, potassium and iron (Lejay et al.,
(miR399) in the phloem acts as systemic signals of low 2003; Hermans et al., 2006; Hammond and White, 2008;
shoot P status (Hammond and White, 2008; Buhtz et al., Liu et al., 2009). However, there are also fine controls
2010; Kragler, 2010; Vance, 2010). Although resupplying specific to particular nutrients. For example, the uptake
P to P-deficient plants ultimately reduces their capacity for rates of P (Drew et al., 1984) and K (Table 2.25) may be
P uptake, this response does not occur immediately (Table more closely related to the concentrations of these nutri-
2.24). Thus, resupplying P after a period of deficiency ents in the shoots than in the roots. Drew and Saker (1984)
can result in greatly increased tissue P concentrations and proposed models for the regulation of K and P uptake in
to P toxicity (Clarkson and Scattergood, 1982; Cogliatti which the fraction of these nutrients delivered to the shoot
and Clarkson, 1983). Such rapid changes in P supply are in excess of demand is translocated back to the roots to
unlikely to occur in soil grown plants. In nutrient solution convey information concerning the nutritional status of the
culture, however, this factor should be considered, espe- shoot. There is good evidence for the cycling of nutrients
cially after the replacement of solutions. within the plant (Section 3.4), and that the translocation
TABLE 2.25 Relationship between the rate of K uptake Rhizosphere Cytoplasm

by maize and the K concentrations of the root and
shoot ATP
K concentration H+
(g kg1dw)
K uptake ADP + P
(pmol cm1 sec1) Root Shoot
15.8 58.5 80.0 FeIII Chelate
NADPH / NADH
28.0 55.5 64.5
Soil
33.8 49.9 43.5 particle
FeIII chelate NADP+ / NAD+
36.8 55.1 41.3
From Barber (1979).
Fe2+ Fe2+
of nutrients and/or their metabolites from the shoot to the

roots in the phloem plays an important role in regulating FIGURE 2.28 Model for root responses to iron deficiency in dicots and
ion uptake by roots. This is the case, for example, for P non-graminaceous monocots (Strategy I): increased acidification of the
(Drew et al., 1984; Drew and Saker, 1984) and K (White rhizosphere by H-ATPases, induction of ferric reductase activity, reduc-
tion of Fe(III)-chelates to Fe2, uptake of Fe2 across the plasma mem-
and Karley, 2010), and for N through nitrate or amino
brane by Fe deficiency-inducible, high-affinity Fe2 transporters.
acids (Cooper and Clarkson, 1989; Liu et al., 2009) and S
through glutathione (Liu et al., 2009; Hell et al., 2010). In
addition, the synthesis of specific microRNAs in shoots of
nutrient-deficient plants, and their translocation to the root confined to the apical zones of growing roots and are fully
in the phloem, could regulate adaptive responses to limited repressed within about one day after resupply of iron.
availabilities of N, S, P, Fe and Cu (Liu et al., 2009; Buhtz Strategy I is typical of dicotyledonous and non-gram-
et al., 2010; Kong and Yang, 2010; Liang et al., 2010; inaceous monocotyledonous plants. It is characterized by
Lundmark et al., 2010; Vance, 2010; Vidal et al., 2010). an increase in ferric (Fe3) reduction capacity, acidifi-
In addition to regulating the uptake capacity for nutri- cation of the rhizosphere, and the release of organic acid
ents, signals of root and shoot nutritional status can also anions and phenolic compounds into the soil solution (Fig.
affect the biomass and morphology of the root system, 2.28). These root responses are closely related to changes
adaptive responses to nutrient deficiency that mobilize in root morphology and anatomy, particularly in the for-
nutrients from recalcitrant compounds, and biochemical mation of transfer cell-like structures in rhizodermal cells.
pathways for the assimilation of nutrients such as N, S and The plasma-membrane ferric (chelate) reductases that cat-
P. For example, plant N status also influences plant shoot/ alyse the reduction of Fe3 to Fe2 in roots of Fe-deficient
root biomass ratio, root morphology and N assimilation and Fe-sufficient plants have similar enzymatic character-
(Hodge, 2004; Hermans et al., 2006; Garnett et al., 2009), istics (Holden et al., 1991), and are encoded by members
plant S status influences plant shoot/root biomass ratio and of the ferric reductase oxidase (FRO) gene family (Puig
modulates sulphate assimilation (Hawkesford and De Kok, et al., 2007; White and Broadley, 2009; Guerinot, 2010).
2006; Hell et al., 2010), and plant P status influences plant Members of the ZIP transporter family, such as the AtIRT1
shoot/root biomass ratio, root morphology, the release of transporter of Arabidopsis thaliana, then mediate Fe2
protons, phosphatases and organic acids into the rhizo- influx to root cells (Puig et al., 2007; White and Broadley,
sphere, and P metabolism in plants (White and Hammond, 2009; Guerinot, 2010). The expression of both AtFRO2
2008). These responses serve primarily to increase the and AtIRT1 in roots of Arabidopsis thaliana vary diur-
acquisition of nutrients and improve the physiological effi- nally, being less at night than in the day, and are increased
ciency by which plants utilize nutrients for growth when under Fe deficiency (Vert et al., 2002). An example of the
they are in short supply. root responses to Fe deficiency, and the corresponding
Regarding their response to Fe deficiency, plants can enhanced rates of Fe uptake, of a Strategy I plant (cucum-
be classified into two categories (Strategy I and Strategy ber) are shown in Table 2.26. The higher reduction rates of
II) (Marschner et al., 1986a, b; Römheld, 1987a,b; Fe3at the outer surface of the plasma membrane of root
Schmidt, 1999; Puig et al., 2007; White and Broadley, rhizodermal cells are correlated with enhanced rates of Fe
2009; Guerinot, 2010). In both strategies, the responses are uptake.
TABLE 2.26 Proton excretion (pH), reducing capacity of the roots and iron uptake ratea in
cucumber (Strategy I) with or without Fe preculture
Fe nutritional Reducing capacity Fe uptake
status Chlorophyll H excretion (µmol FeII g1 (µmol (g1
(preculture) (mg g1 dw) (pH solution) root dw (4 h) 1) root dw (4 h) 1)
Fea 12.2 6.2 3.2 0.03
Fe 7.8 4.8 96.8 2.60

Data compiled from Römheld and Kramer (1983) and Römheld and Marschner (1990).
a
Supply of 1  106 M FeEDDHA, pH 6.2.
COOH COOH COOH Rhizosphere Apoplasm Cytoplasm

H N NH
OH OH
Mugineic acid (MA)
H2C H C MnII FeIII

C O Phytosiderophores
O C O CH2
O C O
Fe
CH2
Soil
H2C N
NH particle
CH2 O
OH CH
CuII FeIIIPS
CH
C O
ZnII
ZnIIIPS CuIIPS
MnIIPS
FIGURE 2.29 Model for root responses to iron deficiency in graminaceous species (Strategy II): enhanced synthesis and release of phytosiderophores
into the rhizosphere, chelation of Fe3, Fe2, Cu2 and Mn2, and transport of metal-phytosiderophore chelates across the plasma membrane by trans-
port proteins. The structures of the phytosiderophore mucigenic acid and its corresponding Fe(III) chelate are also shown.
Strategy II is confined to graminaceous plant species The stability constant for the Fe3-mugineic acid com-
(cereals and grasses) and characterized by an Fe defi- plex in water is in the order of 1033 (Murakami et al.,
ciency-induced enhanced release of non-proteinogenic 1989). The release of phytosiderophores is induced by
amino acids called phytosiderophores (Takagi et al., Fe deficiency (Table 2.27) and rapidly decreases when
1984; von Wirén et al., 1995; White and Broadley, 2009; Fe is resupplied to a Fe-deficient plant (Fig. 2.30A).
Guerinot, 2010). The chemistry of phytosiderophores is Both the release of phytosiderophores and the uptake of
species specific and determines the contrasting abilities metal-phytosiderophore complexes follow a distinct diur-
of different grasses and cereals to acquire Fe (Römheld nal rhythm (Fig. 2.30B) being highest in the first hours
and Marschner, 1990; Bashir et al., 2006; Nagasaka after onset of light. The Fe3-phytosiderophore com-
et al., 2009). Enzymes involved in the synthesis of phy- plex enters the root cytoplasm via proton-coupled Fe3-
tosiderophores from L-methionine include S-adenosyl- phytosiderophore symporters in the plasma membrane of
methionine synthetase, nicotianamine synthase, root cells of cereals and grasses (Fig. 2.29; Römheld and
nicotianamine amino-transferase and deoxymugineic acid Marschner, 1990; von Wirén et al., 1995; Schaaf et al.,
synthase (Bashir et al., 2006; Guerinot, 2010). The expres- 2004; Ishimaru et al., 2006; White and Broadley, 2009;
sion of genes encoding these enzymes, and also of genes Guerinot, 2010). Homologues of the maize yellow stripe 1
involved in S uptake and methionine synthesis, is often (ZmYS1) protein belonging to the oligopeptide transporter
rapidly up-regulated in response to Fe deficiency (Bashir (OPT) family mediate Fe3-phytosiderophore uptake
et al., 2006; White and Broadley, 2009; Guerinot, 2010). by Strategy II plants (von Wirén et al., 1995; Schaaf
Phytosiderophores, such as mugineic acid (Fig. 2.29), et al., 2004; Ishimaru et al., 2006; Puig et al., 2007; White
form highly stable complexes with Fe3, Zn2 and Cu2. and Broadley, 2009; Guerinot, 2010). The corresponding
[µmol Fe equiv. (g root dry wt)−1 h−1]

1.5 Resupply of Fe
9
Release of PS
1.0
6
0.5 3
0 0
10 11 12 13 14 15
Age of plants (days) 0 6 12 18 24
(A) (B) Time of day
FIGURE 2.30 Release of phytosiderophores (PS) from barley roots as affected by plant Fe nutritional status (A), and diurnal rhythm of release of
phytosiderophores (B). Assays were performed on iron-sufficient () and iron-deficient () plants. Data from Römheld (1987a, b) and A. Walter, per-
sonal communication.
graminaceous species. These differences are, as a rule,

TABLE 2.27 Release of phytosiderophores (PS; closely related to the amount and type of the phytosi-
mugineic acid) and uptake of Fe-phytosiderophores derophores released under Fe deficiency (Clark et al.,
in Fe-sufficient and Fe-deficient barley plants 1988; Römheld and Marschner, 1990; Bashir et al., 2006;
PS release Fe uptake Nagasaka et al., 2009). The manipulation of phytosi-
Fe nutritional Chlorophyll (µmol g1 (µmol g1 derophore synthesis, their release into the rhizosphere, and
status concentration root dw root dw the uptake of metal-phytosiderophores by roots offer strat-
(preculture) (mg g1 dw) (4 h) 1) per 4 h) 1) egies to improve Fe uptake by plants and thereby the gen-
Fe 12.8 0.4 0.4 eration of crops for soils with high Fe concentrations but
low Fe availability.
Fe 7.5 8.2 3.4
Römheld and Marschner (1990).
2.5.7 Studying Nutrition at Constant Tissue
Concentration
The common approach of studying plant nutrition in rela-
Zn2 and Cu2-phytosiderophore complexes are also tion to nutrient uptake, nutrient supply, plant nutritional
transported by proton/metal–phytosiderophore symport- status and growth rate has been questioned by Ingestad
ers, but often with a lower affinity and capacity than the and coworkers (Ingestad and Agren, 1992; Ingestad, 1997).
Fe3-phytosiderophore complex (Marschner et al., 1989; These authors have argued that, particularly during the
Ma et al., 1993; von Wirén et al., 1996; Broadley et al., exponential phase of plant growth and even in a flowing
2007; White and Broadley, 2009). The release of phytosi- solution, low external concentrations and plant tissue con-
derophores can also enhance the uptake rate of these met- centrations are difficult to maintain constant. Thus, the rela-
als by increasing their mobility in the rhizosphere (Zhang tionships between external supply, uptake rates and plant
et al., 1991a, b, c). In addition to their ability to take up nutritional status are difficult to ascertain unequivocally.
Fe3-phytosiderophores, graminaceous species can also In order to overcome these difficulties, and also to
take up Fe2 (Ishimaru et al., 2006; Cheng et al., 2007; define the effectiveness of a nutrient in terms of biomass
White and Broadley, 2009; Guerinot, 2010). production per unit of nutrient at different internal con-
Although the phytosiderophore uptake system of centrations, Ingestad and colleagues have used a different
Strategy II plants resembles the siderophore uptake sys- theoretical and experimental approach. In principle, this
tem of microorganisms, its affinity for phytosiderophores approach is based on relative values. To set a constant rela-
is two to three orders of magnitude higher than that for tive uptake, the relative addition rates of nutrients (i.e., the
siderophores such as ferrioxamine B (Bar-Ness et al., supply of nutrients) are divided by the amount of the nutri-
1991, 1992; Crowley et al., 1992), or for synthetic iron ent already in the plant. Accordingly, only the amount of
chelates such as FeEDDHA (Römheld and Marschner, nutrients supplied count, and not the external concentra-
1990; Bar-Ness et al., 1991, 1992). tion. Using this approach, a range of different but constant
Differences exist in ‘Fe efficiency’ (i.e., sensitiv- relative growth rates can be achieved at different degrees
ity to Fe deficiency chlorosis) both between and within of nutrient limitations. Interestingly, although the root/shoot
TABLE 2.28 Uptake and translocation of K (42K) and Ca (45Ca) supplied to different zones of the
seminal roots of maizea
Root zone supplied
(distance from tip, cm)
Nutrient
(1 mM) Accumulation and translocation 0–3 6–9 12–15
K Translocation to shoot 3.8 14.6 15.6
Accumulation in zone of supply 11.5 3.8 1.9
Translocation to root tip – 4.3 2.0
Total 15.2 22.7 19.5
Ca Translocation to shoot 2.4 2.2 2.4
Accumulation in zone of supply 4.1 1.6 0.4
Translocation to root tip – – –
Total 6.5 3.8 2.8
Based on Marschner and Richter (1973).
a
Data expressed as micromoles per 12 plants in 24 h.
dw ratio of the nutrient-limited plants is often large, visual cells in the basal zones. The apical root zones have higher
deficiency symptoms are absent. respiration rates (Thomson et al., 1989b), which decrease
This highly formal concept is an interesting variation to rapidly when the carbohydrate supply to roots is inter-
the common approach for studying the nutrition of plants, rupted, for example following excision (Brouquisse et al.,
in which the influence of external concentration and plant 1991). In general, there is a tendency for the rate of ion
nutrient status on nutrient uptake, growth responses and uptake per unit root length to decrease with distance from
various physiological and biochemical parameters (e.g., the root apex. However, this tendency strongly depends
photosynthesis) are studied. This concept allows study- on the identity of the ion, plant nutritional status and plant
ing the effects of mineral nutrition under suboptimal but species. When K or Ca are supplied to different regions of
steady-state conditions. However, these steady-state con- seminal roots of maize (Table 2.28), the uptake rate of K is
ditions, in which the relative nutrient supply is adjusted slightly lower in the apical zone than the sub-apical zone,
to the relative growth rate, are not typical of those expe- despite the high K requirement for growth. The high K
rienced by plants growing in the field. For field grown concentration in root apical cells of about 200 mM (Huang
plants, fluctuations in nutrient supply to the roots are as and Van Steveninck, 1989a) is maintained not only by
common as fluctuations in other environmental parameters uptake from the external solution but also by delivery from
such as irradiation, temperature and water supply. To cope more basal root zones (Table 2.28) or from the shoot via
with these fluctuations plants possess a range of adaptive the phloem (Gould et al., 2004). Similar observations have
mechanisms. Fluctuations in nutrient supply are compen- been made in other cereals (White et al., 1987; Vallejo
sated for by modulating uptake capacity, changes in root et al., 2005) and also in non-mycorrhizal long roots of
morphology and physiology and root/shoot biomass ratio perennial plant species such as Norway spruce (Häussling
(Chapters 14–16), and the storage and remobilization of et al., 1988).
mineral nutrients (Chapters 3 and 6). In contrast to K the uptake of Mg, and particularly of
Ca, is higher in apical than in basal root zones (Marschner
2.6 UPTAKE OF IONS AND WATER ALONG and Richter, 1973; Ferguson and Clarkson, 1976;
Häussling et al., 1988; White, 2001). This is also shown
THE ROOT AXIS in Table 2.28. Because Ca mobility in the phloem is low,
Roots vary both anatomically and physiologically along apical cells of the root must meet their Ca demand for
their longitudinal axes. This should be borne in mind when growth by direct uptake from the external solution. Root
models for ‘the’ behaviour of root tissue and root cells apical zones also contribute considerably to Ca delivery
are based on studies with isolated roots or roots of intact to the shoot (Table 2.28; Clarkson, 1984; White, 2001).
plants. In the apical zone, non-vacuolated cells dominate. At the root tip, Ca may reach the xylem through an exclu-
These cells differ in many respects from the vacuolated sively apoplasmic pathway or may be transported across
TABLE 2.29 Rate of P uptake by various root zones of

barley plants after different pre-treatment
Lateral
P uptake rate (pmol mm3 (24 h)1) roots
Root zone (distance from

root tip, cm)
Pretreatment
for 9 days 1 2 3
With P 2,019 1,558 970 Root
hairs
Without P 3,150 4,500 4,613
Based on Clarkson et al. (1987).
Mucilage
FIGURE 2.31 Schematic representation of anatomical changes along

the Casparian band through immature, unsuberized endo- the axis of a maize nodal root. In basal zones there is degeneration of cor-
dermal cells (White, 2001; Moore et al., 2002). Calcium tical cells and formation of tertiary endodermis.
delivery to the xylem is also high in basal root zones,
where lateral roots emerge from the pericycle, disrupt-
ing the integrity of the Casparian band (Clarkson, 1984; or P supply (He et al., 1992; Lynch, 2007). Despite these
White, 2001). The apoplasmic pathway is also important anatomical changes, the basal root zones still have a con-
for the movement of Na, Zn, Fe and Cd to the xylem (Yeo siderable capacity for ion uptake (Drew and Fourcy, 1986)
et al., 1987; Taiz and Zeigler, 2006; Broadley et al., 2007; and also for radial transport, indicating that the strands of
Plett and Møller, 2010; Lux et al., 2011). The delivery of cells bridging the cortex maintain sufficient ion transport
these elements to the xylem is often greatest at the root tip. capacity from the rhizodermis to the endodermis (Drew
The decline in P uptake along the root axis is much and Saker, 1986).
less striking than that for Ca (Ferguson and Clarkson, Water uptake can affect ion uptake both directly,
1975; Clarkson et al., 1978; Rubio et al., 2004). In soil- through effects on the rate of radial transport of ions
grown maize this decline is mainly related to a decrease in through the apoplasm, and indirectly, by influencing
root hair viability and, thus, in absorbing root surface area the supply of ions to the plasma membrane of root cells.
(Ernst et al., 1989). The gradient in P uptake along the root Water uptake is usually low at the extreme root apex, but
axis also depends on the P nutritional status of the plant increases in the elongation zone and reaches a maximum
and may be reversed under deficiency in favour of the in the root hair zone, where the endodermis is undergo-
basal zones (Table 2.29). The situation is different under ing suberization (e.g., Sanderson, 1983; Boyer, 1985;
Fe deficiency in Strategy I plants where the apical, but not Häussling et al., 1988). Water uptake is often reduced
the basal, root zones increase their capacity for Fe uptake strongly following suberization of the endodermis and,
by a factor of up to 100 (Römheld and Marschner, 1981b). particularly, the exodermis. Water can reach the xylem
Apical, or immediately sub-apical, root zones generally through both the apoplast and via root cells (Steudle,
contribute most to nitrate and ammonium uptake by intact 2000). Transport through root cells is facilitated by
plants irrespective of their nutritional status, although the aquaporins (Javot and Maurel, 2002; Hachez et al., 2006;
magnitude of the decline in uptake with distance from the Maurel et al., 2008). Aquaporins are found in various
root apex depends greatly on root anatomy (Reidenbach membranes of root cells, including the plasma membrane
and Horst, 1997; Colmer and Bloom, 1998; Sorgona et al., and the tonoplast. Recent data, using mercury to inhibit the
2010). Indeed, it should be noted that the uptake of most activity of aquaporins, suggest that rapid changes in root
elements is restricted when the rhizodermis and cortex hydraulic conductivity in response to many stimuli, such
cells of basal (older) regions of the roots collapse and die. as diurnal cycles, nutrient deficiency, salt stress, low tem-
Formation of cortical gas spaces (aerenchyma) par- peratures, anoxia and drought, are the result of changes
ticularly in more basal root zones can often be observed in cell membrane permeability achieved by regulation of
(Fig. 2.31). The formation of aerenchyma is a typical aquaporin activity (Javot and Maurel, 2002; Maurel et al.,
response to oxygen deficiency in the root zone in plant 2008). The abundance of aquaporins is often greatest in the
species adapted to wetland conditions (Section 16.4), but elongation and mature root zones (Hachez et al., 2006).
it can also be induced, for example, in maize roots under In these root zones, strong expression of genes encoding
fully aerobic conditions by temporary deprivation of N aquaporins is observed in the endodermis and exodermis,
Cortex root hair cells, play a key role in the acquisition of min-
eral nutrients, especially K and P (Gahoonia and Nielsen,
Stele
2004; Gahoonia et al., 2006; Jung et al., 2009; Zhu et al.,
A
2010), the relative importance of the two pathways for
Early metaxylem
solute transport across the root cortex is unknown. It will
Late metaxylem depend on: (i) the external concentration versus the capac-
Root ity and affinity of the transport system for a particular
hairs Phloem
Endodermis solute at the plasma membrane of root cells; (ii) the root
Casparian band zone considered: depending on environmental conditions
and the growth rate of the root, the exodermis can develop
Exodermis
B (hypodermis) within a centimetre of the root apex or remain undeveloped
Rhizodermis (Ma and Peterson, 2003) and may possess ‘passage cells’
(Storey and Walker, 1987); and (iii) the hydraulic con-
FIGURE 2.32 Segment of a transverse section of a maize root showing ductivity of the root zone considered and the transpiration
(A) symplasmic and (B) apoplasmic pathways of solute movement across rate of the shoot. For water, estimates of the contribution
the root.
of the apoplasmic pathway to radial transport across roots
vary between about 10 and 70% (Javot and Maurel, 2002;
presumably to allow water to bypass the Casparian band Hachez et al., 2006; Maurel et al., 2008).
through a transcellular pathway (Hachez et al., 2006) The endodermis is also not a perfect barrier to the apo-
plasmic movement of water and solutes from the cortex
2.7 RADIAL TRANSPORT OF IONS AND to the stele (Fig. 2.32). In addition to the presence of pas-
sage cells in some plant species, this barrier may be ‘leaky’
WATER ACROSS THE ROOT
at two sites along the root axis, at least. At the root apex,
There are two parallel pathways of movement of solutes where the Casparian band is not yet fully developed, the
and water across the cortex towards the stele: one pass- apoplasmic movement of water and solutes to the stele can
ing through the apoplasm (cell walls and intercellular occur. However, the movement of some solutes, such as
spaces) and another passing from cell to cell in the sym- polyvalent cations like aluminium, through the apoplasm
plasm through the plasmodesmata (Fig. 2.32). In most of of the root apex can be restricted by mucilage formed at
the root, the apoplasmic movement to the stele is restricted the external surface of the rhizodermal cells (Section
by the Casparian band in the walls of endodermal cells 15.4). The apoplasmic pathway to the stele is also possi-
(White, 2001). This band is suberized and joins each endo- ble in basal root zones where the structural continuity of
dermal cell (stage I endodermis). In the basal regions of the endodermis is disrupted transiently by the emergence
the root, suberin lamellae cover the entire surface of endo- of lateral roots from the pericycle, as has been demon-
dermal cells (stage II endodermis). This prevents endo- strated, for example, for Ca (White, 2001), Al (Rasmussen,
dermal cells taking up solutes from the apoplasm (Moore 1968) and water (Häussling et al., 1988; Wang et al.,
et al., 2002). Thick cellulose secondary walls are deposited 1991). This ‘bypass-flow’ becomes particularly impor-
over the suberin lamellae, which can be lignified (stage tant for water supply to the shoot at high transpirational
III endodermis). The nature and extent of these cell wall demand (Sanderson, 1983) and in the accumulation of Na
modifications are determined by both genetic and environ- in leaves under saline conditions (Yeo et al., 1987; Plett
mental factors. and Møller, 2010). Both genetic and environmental factors
In most angiosperms, another apoplasmic barrier, the influence the movement of water and solutes via the apo-
exodermis, can develop in parallel with the endodermis plasmic pathway through their effects on the development
(Ma and Peterson, 2003). The exodermis develops in the of the endodermis and exodermis. Accelerated deposition
same three stages as the endodermis. Formation of an exo- of suberin and lignin restricts the apoplasmic movement of
dermis is found, for example, in Zea mays, Allium cepa, or cations and other solutes to the xylem (White, 2001;
Helianthus annuus, but not in Vicia faba or Pisum sativum Enstone et al., 2002; Krishnamurthy et al., 2009; Lux
(Enstone and Peterson, 1992). However, there are some- et al., 2011) and reduces hydraulic conductivity (Boyer,
what different views on the function of the exodermis as 1985; Cruz et al., 1992).
an effective barrier for transport of water and solutes in the The symplasmic pathway plays a key role in deliver-
apoplasm of the root cortex. Termination of the apoplasmic ing most nutrients to the xylem, beginning either at the
pathway at the exodermis, as suggested by Enstone and rhizodermis and the root hairs, at the exodermis, or at the
Peterson (1992), would confine the entry of solutes and endodermis. Radial transport in the symplasm requires
water to the root symplast to the rhizodermal cells in basal movement through plasmodesmata, which connect neigh-
root zones. Although rhizodermal cells, and in particular bouring root cells (Fig. 2.33). Plasmodesmata have a
(A) (B)
Plasma TABLE 2.30 Intracellular K activity and number of
membrane
plasmodesmata in tangential walls of hair and hairless
Proteins cells of the root epidermis
Number of
ER Microchannel
plasmodesmata
Cytosol
Plant K activity Per cell
Callose species Cell type (mM) Per µm2 junction
Protein
Trianea Hair 133 2.06 10,419
Cell wall bogotensis
Hairless 74 0.11 693
Cytoplasmic sleeve Raphanus Hair 129 0.16 273
Desmotubule sativus
Hairless 124 0.07 150
Plasma membrane
From Vakhmistrov (1981).
ER
FIGURE 2.33 Schematic representation of plasmodesmata including High cytosolic Ca2 concentrations induce closure of
substructural components. Solute fluxes between adjacent cells occur in plasmodesmata (Tucker, 1990) and many environmental
the cytoplasmic sleeve, between the plasma membrane and the appressed stimuli that increase cytosolic Ca2 also disrupt the sym-
endoplasmic reticulum (ER) forming the desmotubule. Partial control plasmic movement of water and nutrients across the root.
of solute fluxes by callose deposition in the cell wall. The cytoplasmic
sleeve is interrupted by actin and other proteins that create microchannels
The number of plasmodesmata per cell varies consider-
through which solutes can diffuse. Modified from Maule (2008). ably between plant species and cell type (Table 2.30).
Rhizodermal cells that have developed into root hairs
generally have more plasmodesmata than other rhizoder-
mal cells. The relatively small number of plasmodesmata
complex structure (Lucas and Lee, 2004; Maule, 2008; in Rhaphanus raises the question as to whether the root
Lucas et al., 2009). The simplest type, which occurs in hairs are of major importance for symplasmic radial trans-
young tissues, comprises a tube of appressed endoplasmic port in this plant species. However, not only the number
reticulum (ER) running through the pore, the desmotubule. of plasmodesmata, but also whether they are functional
The transport of solutes and water between cells occurs must be taken into account. In the endodermis of young
in the ‘cytoplasmic sleeve’, i.e. the cytosol between the barley roots, on average 20,000 plasmodesmata per cell
desmotubule and the plasma membrane (Fig. 2.33). Protein have been found (Helder and Boerma, 1969). In the terti-
structures in the cytoplasmic sleeve create microchannels ary (lignified) endodermis of older zones of barley roots,
through which solutes can diffuse (Lucas and Lee, 2004; there are far fewer plasmodesmata, but the number appears
Maule, 2008; Lucas et al., 2009). In more mature tissues, to be sufficient to permit considerable radial transport of
the structure becomes more complex through the addition both water and ions through the endodermis (Clarkson
of branches and the formation of central cavities (Maule, et al., 1971).
2008; Lucas et al., 2009). Plasmodesmata can be closed The mechanism of symplasmic transport of solutes
and opened by the production and degradation of a ‘col- seems to be chiefly by diffusion, facilitated by radial water
lar’ of callose (β-1,3-glucan) and they generally have a size flux and cytoplasmic streaming. During their radial trans-
exclusion limit of about 1 kDa, which is regulated physi- port through the symplasm, elements can be metabolized
cally by the collar and also by interactions with cytosolic and/or sequestered in the vacuoles of root cells. When a
proteins (Lucas and Lee, 2004; Maule, 2008; Lucas et al., nutrient is supplied to roots of a plant that is deficient in
2009). Indeed, plasmodesmatal microchannels can dilate that nutrient (‘low-salt’ roots), it is accumulated in vacu-
to allow the passage of solutes in excess of 20 kDa. The oles of root cells resulting in an immediate accumulation
primary role of plasmodesmata appears to be cell com- in roots and a delay in its translocation from the roots to
munication, as they regulate the transport of transcription the shoots (Fig. 2.34). Thus, when the supply of a nutrient
factors and microRNAs that control plant development and is suboptimal, the roots usually have higher tissue concen-
responses to biotic and environmental challenges (Lucas et trations of that particular nutrient than the shoot. In long-
al., 2009). Addionally, the regulation of plasmodesmatal term studies, this phenomenon is responsible, in part, for
conductance represents another mechanism of cellular the often observed shift in the relative growth rates of roots
control of ion fluxes across the root. and shoots in favour of the roots under nutrient deficiency.
(A) (B)
ving)
12 closed (li
open Soils heath
Roots Large metaxylem
content (µmol plant−1)
10 vessels
Nodal roots
8
Shoots
Primary root
6
Shoots
4
Roots
FIGURE 2.35 Model of root hydraulic conductivity and formation of a
42K
2 soil rhizosheath in the root system of maize. Modified from Wenzel et al.
(1989).
0
0 2 4 6 8 10 12 0 2 4 6 8 10 12
Time (hours) Time (hours)
delay in metaxylem maturation not only affects hydraulic
FIGURE 2.34 Accumulation and translocation rates of K (42K) in bar-
ley plants from a solution containing 1 mM KCl (0.5 mM CaSO4) after conductivity of the roots and plant water relations (Wang
preculture with (A) or without (B) 1 mM KCl. et al., 1991) but also the movement of solutes to the xylem
and their translocation to the shoot.
The vacuoles of root cells also remove potentially

2.8 RELEASE OF IONS INTO THE XYLEM
toxic elements from the symplasmic pathway. For exam-
ple, vacuolar sequestration of Na in the root accounts for After radial transport through the symplasm to the stele,
the restricted shoot transport of Na in natrophobic plant ions and organic solutes (amino acids, organic acids) are
species (Chapter 3). Preferential accumulation in roots released into the xylem. This release (xylem loading)
also restricts the translocation of Ca, Mo, Cd and Al to the into fully differentiated, non-living xylem vessels occurs
shoot (Conn and Gilliham, 2010). In contrast, in plants suf- across the plasma membrane of xylem parenchyma cells.
ficiently supplied with P, symplasmic transport of P, and The membrane potential of these cells is slightly nega-
its translocation to the shoot, is greater than accumula- tive (Bowling, 1981) and the xylem sap has a pH between
tion in root vacuoles (Lamaze et al., 1987). The exchange about 5.2 and 6.0 (Section 3.2). Solutes enter the xylem
rate between ions in the vacuoles of cortex cells and those through ion channels or uniporters, if their electrochemi-
in the symplasm depends on the ion species (K  Na; cal gradients allow this, or their transport is coupled to the
NO3  SO42), and the half-time for exchange is gener- proton electrochemical gradient generated by the plasma
ally in the order of at least a few days. membrane H-ATPase or directly to ATP hydrolysis. The
The radial transport of water and solutes is strongly xylem parenchyma cells are also responsible for the reab-
influenced by maturation of the xylem vessels along the sorption of solutes from the xylem sap by tissues along the
root axis. For example, in graminaceous species such as pathway to the shoot.
maize growing in soil, two root zones can be observed: The key role of the H-ATPases in the plasma mem-
‘sheathed’ zones which are covered by a layer of strongly brane of parenchyma cells in xylem loading is now well
adhering soil, the rhizosheath and ‘bare’ zones (Fig. 2.35). established. Protons are pumped into the xylem both to
The development of the rhizosheath appears to be related generate a negative membrane potential and to acidify the
to the presence of root hairs (Haling et al., 2010). In the xylem sap (De Boer and Volkov, 2003). The K electro-
sheathed zones, the metaxylem vessels are still alive and chemical gradient is sufficient for K to be loaded into the
non-conducting, whereas in the bare zones the metaxy- xylem by voltage-gated, outwardly rectified K-channels,
lem is mature (McCully and Canny, 1988). Accordingly, such as the AtSKOR K-channel of Arabidopsis thaliana
the hydraulic conductivity of bare roots is about 100 times (Gaymard et al., 1998). Similarly, anion channels can
greater than that of sheathed roots (Wenzel et al., 1989). facilitate the movement of nitrate, sulphate, phosphate and
This difference in hydraulic conductivity and thus water chloride from the symplasm to the xylem in the direction
uptake results in high water contents in the rhizosphere of their electrochemical gradients (White and Broadley,
soil of the sheathed zones and low water contents in the 2001; Köhler et al., 2002; Gilliham and Tester, 2005). In
rhizosphere soil of the bare zones. Living metaxylem ves- addition, nitrate can be loaded into the xylem by mem-
sels can be found up to 20–30 cm proximal to the root tip bers of the NRT1 (nitrate transporter 1) family (Lin et al.,
in maize (Wenzel et al., 1989), and up to 17 cm proximal 2008). Cations present at low concentrations in the root
to the root tip in soybean (Kevekordes et al., 1988). This symplasm are loaded into the xylem by active transport
TABLE 2.31 Root uptake and translocation to the shoot of phosphate and sulphate in the wildtype
and pho1 mutant of Arabidopsis thaliana
Phosphatea Sulphate
Root uptake Translocation Root uptake Translocation
Genotype (nmol g1 h1) to shoot (%) (nmol g1 h1) to shoot (%)
Wildtype 1,593 35 291 25
Mutant 1,559 0.9 367 12
Poirier et al. (1991).
a
Supply of 8 µM Pi.
mechanisms. Members of the P2A and P2B Ca2-ATPase are up to 400 mM. Upon maturation of the metaxylem ves-
families load Ca into the xylem and members of the heavy sels, the accumulated K, together with the other solutes
metal P1B-ATPase family load Zn and Cu into the xylem in the vacuoles, is released into the transpiration stream.
(White and Broadley, 2003, 2009). It is thought that Mg According to McCully and Canny (1988), this leakage
and Mn are also loaded into the xylem by ATPases, from maturing xylem vessels could account for about 10%
although the genes encoding these transporters are not yet of the shoot demand of growing maize plants. Thus, a sig-
known. Boron is loaded into the xylem by orthologues of nificant proportion of the solutes present in the xylem sap
the Arabidopsis AtBOR1 transporter (Miwa and Fujiwara, (including proteins) may derive not from active xylem
2010). The regulation of xylem loading separately from loading but from maturing xylem vessels.
solute uptake offers additional possibilities to control the
selectivity and rate of long-distance transport to the shoot, 2.9 FACTORS AFFECTING ION RELEASE
for example in response to shoot demand. INTO THE XYLEM AND EXUDATION
Separate genetic control of solute uptake and xylem
loading from that of root cortex cells is in agreement with
RATE
the observation that selective inhibitors of protein syn- The permeability of plant membranes to water is higher
thesis strongly impair xylem loading of nutrients, such than that to ions. Plant cells or roots therefore behave as
as K, without affecting their accumulation in the roots osmometers. Ion release into the apoplasm of the stele
(Läuchli and Pflüger, 1978; Morgan et al., 1985) and that decreases both the osmotic potential and the water poten-
diurnal fluctuations in nutrient uptake by the roots and tial (they become more negative) in the stele, and a cor-
their delivery to the xylem do not coincide (Herdel et al., responding net flux of water from the external solution is
2001). Another example of the separate genetic control induced. As a result of this water flux, the hydrostatic pres-
of solute uptake and xylem loading is shown in Table sure increases. As the endodermis with its Casparian band
2.31. Compared with the wildtype, the pho1 mutant of ‘seals’ the apoplasm of the stele, the hydrostatic pressure in
Arabidopsis thaliana requires very high external P supply the stele induces a volume flow of water and solutes in the
for normal growth. The recessive gene (pho1) regulates the non-living xylem vessels towards the shoot. Because of this
loading of P into the xylem (Poirier et al., 1991). At low P ‘root pressure’ droplets are sometimes released on the tips
supply, the mutant becomes severely P deficient because of and margins of leaves, a process known as guttation. This is
impaired translocation of P to the shoot, although P uptake particularly apparent in seedlings and young plants at night
by the roots does not differ from the wildtype plant (Table and in the early morning (under conditions of high relative
2.31). Sulfate translocation to the shoot is similar in the air humidity and low transpiration). Exudation from the
mutant and the wildtype plant. Evidence for tight regula- stumps of cut plants (e.g., freshly mown grass) is also the
tion of P loading into the xylem is also shown by the ina- result of root pressure. Root pressure and the correspond-
bility of maize plants to meet the P demand of the shoot at ing volume flow in the xylem are of particular importance
low root zone temperatures (Engels and Marschner, 1992a). for the long-distance translocation of Ca into low-transpir-
The discovery of the abundance of living metaxy- ing organs such as fruits. Volume flow and composition of
lem vessels in more than half of the total root length in the xylem exudate can provide important information on
mature maize plants (Wenzel et al., 1989) renewed the the influence of external and internal factors on root activ-
view of leakage as a mechanism of ion release into the ity and metabolism, nutrient uptake and assimilation in the
xylem (McCully et al., 1987). The concentrations, of K, roots, release into the xylem and the cycling of nutrients and
for example, in the vacuoles of living metaxylem vessels organic solutes in plants.
TABLE 2.32 Relationship between external concentration, exudate concentration, and exudate volume
flow in decapitated sunflower plants
External solution Exudate (mM) Concentration factor Exudation
KNO3  CaCl2 volume flow
(mM each) K Ca2 NO3 K Ca2 NO3 (mL (4 h)1)
0.1 7.3 2.8 7.4 73 28 74 4.0
1.0 10.0 3.2 10.7 10 3.2 10.1 4.5
10.0 16.6 4.2 10.3 1.7 0.4 1.0 1.6
TABLE 2.33 Exudate volume flow and K and Ca concentrations in the exudate of
decapitated maize plants at different temperatures
Exudation
concentration (mM)
Exudate volume Ratio K/
Temperature (°C) flow (mL h1) K Ca2 Ca2
8 1.32 13.4 1.5 8.9
18 5.48 15.2 1.0 15.2
28 7.93 19.6 0.8 24.5
Marschner (1995).
a
Concentration of KNO3 and CaCl2 in the external solution: 1 mM each.
For technical reasons, it is difficult to measure ion xylem exudate. However, the relative concentration differ-
release into the xylem directly. Although secretions from ence decreases as the external concentration is increased
xylem-feeding insects, such as the meadow spittlebug (Table 2.32). Thus, the concentration gradient (‘concentra-
(Philaenus spumarius), can be obtained from intact plants tion factor’) between the external solution and the xylem
(e.g., Watson et al., 2001; Malone et al., 2002; Teakle exudate decreases, and can even fall below 1 in the case
et al., 2007), most experimental evidence on xylem load- of Ca, i.e. the concentration of Ca in the xylem exudates is
ing comes from studies of xylem exudate, or xylem sap, lower than that in the external solution. The volume flow
obtained from isolated roots or decapitated plants (Section of xylem exudation shows a somewhat different pattern,
3.2). Because of reabsorption along the xylem pathway and is maximal at an external concentration of 1.0 mM
(Section 3.2), and the contribution of solutes from matur- in the experiment reported in Table 2.32. At 0.1 mM, this
ing metaxylem vessels, the concentration of ions at the flow is limited by the ion concentration in the xylem. In
sites of collection can differ from that at the sites of load- contrast, at 10.0 mM, the flow is limited by water availa-
ing into the non-living xylem vessels. When interpreting bility (i.e., the low water potential in the external solution)
analyses of xylem exudate it should be kept in mind that (i) and the small concentration gradient between the external
at least two separately regulated membrane transport proc- solution and the xylem. The increase in the concentration
esses are involved in symplasmic radial transport of nutri- of nutrients in the xylem exudate with increasing exter-
ents from the external solution into the xylem (i.e., influx nal concentration from 1.0 to 10.0 mM does not compen-
to the symplasm and xylem loading), (ii) an apoplasmic sate for the decrease in the exudation volume flow. Thus,
pathway can contribute to the delivery of water and solutes in contrast to their accumulation in roots, which generally
to the xylem, and (iii) xylem sap volume flow is affected follows a hyperbolic relationship with the external con-
by root hydraulic conductivity and rate of transpiration. centration, the rate of root pressure-driven translocation
of nutrients to the shoot can decline at high external solute
2.9.1 External and Internal Factors Affecting concentrations due to limited water uptake.
An increase in the root zone temperature often has a
the Composition of Xylem Sap greater effect on the exudation volume flow than on the
As a rule, an increase in the external ion concentration ion concentrations in the exudate (Table 2.33). This is
leads to an increase in the concentration of ions in the consistent with the expectation that a root behaves as an
TABLE 2.34 Exudation volume flow and ion TABLE 2.35 Flow rate and ion concentration in the
concentration in the exudate of decapitated maize xylem exudate of wheat seedlingsa
plants with (O2 treatment) or without root respiration
Treatment
(N2 treatment)
Parameter KNO3 K2SO4
Exudate
Exudation volume concentration Exudation flow rate 372 180
Treatmenta,b flow (ml h1) (mM) (µl h1 50 plants1)
K Ca2 Ion concentration (mM)
O2 8.83 16.6 1.8 Potassium 23.3 24.5
N2 1.90 15.2 1.7 Calcium 9.1 9.5
Marschner (1995). Nitrate 18.1 0.0

a
Concentration of KNO3 and CaCl2 in the external solution: 0.5 mM
each. Sulphate 0.2 0.8
b
Respiration treatment consisted of bubbling oxygen or nitrogen through
the external (nutrient) solution. Organic acids 9.6 25.8
From Triplett et al. (1980).
a
Seedlings were supplied with either KNO3 (1 mM) or K2SO4 (0.5 mM) in
the presence of 0.2 mM CaSO4.
osmometer. Temperature has a marked effect on the rate of

solute uptake, transport through the symplasm and release
into the xylem, and water moves accordingly along the between the treatments (Table 2.35). When plants are sup-
water potential gradient. Since different transport proteins plied sulphate rather than nitrate, the difference in negative
facilitate the movement of each nutrient across the plasma charge in the exudate is approximately compensated for by
membrane of root cells, and the relative contributions of elevated concentrations of organic acid anions. However, the
symplasmic and apoplasmic transport differ between nutri- capacity of the roots to maintain the cation–anion balance
ents, temperature can have differential effects on the deliv- by organic acid synthesis in the K2SO4 treatment appears to
ery of each element to the xylem. For example, an increase be limited, which leads to a decrease in the rate of K and
in the root temperature results in an increase in K concen- Ca release into the xylem and a corresponding decrease in
tration but in a decrease in Ca concentration of the exudate. exudation flow rate when compared to the KNO3 treatment.
This shift in the K/Ca ratio may reflect temperature effects Because of the energy demand for ion transport proc-
either on membrane selectivity or on the relative impor- esses, release of ions into the xylem and the correspond-
tance of the apoplasmic pathway of radial transport of Ca ing changes in root pressure are also closely related to the
and water (Engels et al., 1992). Similar shifts in the K/Ca carbohydrate status of the roots (Table 2.36). Variation in
transport ratio are also observed at different soil tempera- the length of the photoperiod one day before decapitation
tures (Walker, 1969). This temperature effect may have affected the carbohydrate status of roots and, correspond-
important implications for the Ca nutrition of plants. ingly, the rate and duration of exudation volume flow after
The rate of xylem loading is closely related to root res- decapitation. Both the uptake and transport rate of K in
piration (Table 2.34). A lack of oxygen strongly reduces roots with high carbohydrate content are greater than in
exudation volume flow but not the concentrations of K and roots with low carbohydrate content. The higher trans-
Ca in the exudate. Oxygen deficiency seems to affect the port rate is closely related to the exudation volume flow.
release of ions into the xylem and root hydraulic conduc- In roots with low carbohydrate content, reserves are rap-
tivity to the same degree. idly depleted after decapitation and there is a correspond-
The cation–anion balance in the xylem exudate needs ing decline in the rate of exudation volume flow within 8 h.
to be maintained. Thus, the accompanying ion can affect This depletion of carbohydrates in the roots of decapitated
xylem exudation flow and sap composition even at low plants and the consequent decline in xylem exudation flow
rhizosphere concentrations (Table 2.35). When KNO3 is is one of the factors restricting studies of xylem loading.
supplied, the exudation flow rate is almost twice as high as Solute fluxes into the xylem and exudation volume flow
the flow rate when an equivalent concentration of K2SO4 is also exhibit endogenously regulated diurnal fluctuations.
added. Since the K concentration in the exudate is the same These are maintained in plants transferred to continuous
in both treatments, the translocation rate of K supplied as darkness (Ferrario et al., 1992a, b) but disappear in plants
K2SO4 is only about half the rate of K supplied as KNO3. maintained under continuous light (Herdel et al., 2001).
In contrast to the K concentration, the concentrations of These phenomena are related not only to the carbohydrate
nitrate and sulphate in the exudate exhibit large differences status of the roots but also to plant nutritional status.
TABLE 2.36 Relationship between photoperiod, TABLE 2.37 Role of shoot demand on net uptake,
carbohydrate content of roots, and uptake and net translocation and flux of K in the xylem exudate of
translocation of K in decapitated maize plantsa maize
Photoperiod (h) K flux (µmol g1 root fw h1)
12/12b 24/0 Net Xylem
Shoot Net uptake translocation exudates
Carbohydrate in roots (mg) 122 (48) 328 (226)c
demanda 0–3 days 0–3 days day 3
Total potassium uptake (mmol) 1.3 5.0
High 2.26 1.83 8.55
Potassium translocation in 1.0 3.5
Low 2.28 1.17 2.46
exudation volume flow (mmol)
Engels and Marschner (1992b).
Exudation volume flow (mL 8 h1) 30.3 88.5 a
Shoot demand altered by the shoot base temperature, see Table 2.12.
Relative decline in flow rate 60 12

within 8 h (%)
Marschner (1995).
a
Data per 12 plants.
b
Hours of light/hours of darkness. This pretreatment with different day
lengths was for one day (i.e., the day prior to decapitation). Malone et al., 2002; Teakle et al., 2007), but this technique
c
Numbers in parentheses denote carbohydrate content after 8 h is rarely applied. Alternatively, xylem volume flow can be
(decapitation).
increased artificially by increasing the external pressure in
the root zone (pressure chamber) or by collecting exudates
from the cut stump under a vacuum. Using either method,
2.9.2 Xylem Exudate, Root Assimilation and xylem volume flow is increased and, hence, concentrations
of nutrients are usually decreased. The estimated translo-
Cycling of Nutrients cation rates to the shoot can differ between these methods
Analyses of xylem exudates provide valuable information and can also be quite different from the results using intact
about assimilation of nutrients in the roots, for example plants (Salim and Pitman, 1984; Allen et al., 1988; White,
the capacity of roots for nitrate reduction or N2 fixation. In 1997b). Furthermore, irrespective of the collection method,
soybean and other tropical legumes, the proportion of urei- the xylem sap also contains shoot-derived nutrients recy-
des (Chapter 16) to total N in the xylem exudate reflects cled in the phloem and reloaded into the xylem in the roots
nodule activity and is also a suitable indicator in field- (Section 3.4). The recirculation of water may also have to
grown plants of the relative contribution of N2 fixation to be considered in interpreting analyses of xylem sap (Tanner
the N nutrition of legumes (Peoples et al., 1989). In non- and Beevers, 1990). The fractions of recycled nutrients can
legumes, the forms of N in xylem exudate can provide be particularly high in the cases of K, P, N and S (Section
information on its assimilation by roots and the relative 3.4), and may lead to misinterpretation of, for example, the
importance of various organic and inorganic fractions in its capacity of roots to reduce nitrate and sulphate.
long-distance translocation in the xylem (Van Beusichem The proportion of recycled nutrients in the xylem sap
et al., 1988). Similarly, studies of the chelation of heavy depends on various factors, such as plant species and nutri-
metals in xylem sap have increased our understanding of tional status in general and shoot demand in particular, as
their translocation within the plant, and the transport pro- shown for K in Table 2.37. When shoot demand is high, K
teins involved (Section 3.2). Analyses of xylem exudate can translocation in the xylem exudate increases greatly and net
also inform about the metabolism and translocation of hor- translocation is also higher, but net uptake is unaffected.
monal signals from roots to shoots (Chapters 3 and 5). The Accordingly, root K concentration is lower in the plants
discovery of unexpectedly high concentrations of sugars in with high shoot demand. The differences between net trans-
xylem exudates of annual species provided insight to their location and xylem transport of K reflect differences in the
carbon economy (Cataldo et al., 1988; Canny and McCully, recirculation of K which, against expectation, was higher in
1989). plants with high shoot demand. Recycling of K to the roots
There are, however, several factors that need to be con- in plants with high shoot demand may be explained by the
sidered in the interpretation of such analyses. Xylem sap role of K in xylem transport of nitrate (Section 3.4). This is
collected from decapitated plants represents only the root an example of the important insight xylem sap analyses can
pressure component of xylem volume flow. For evalua- provide when combined with other measurements, such as
tion of the transpirational component, xylem sap should be rates of net uptake and accumulation in plant tissues or the
collected from intact plants in situ. This can be achieved analysis of phloem sap, to the regulation of xylem loading
using xylem-feeding insects (e.g., Watson et al., 2001; and the recycling of nutrients within the plant.

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Ion Uptake Mechanisms of Individual Cells

Marschner’s Mineral Nutrition of Higher Plants. DOI: 10.1016/B978-0-12-384905-2.00002-9

TABLE 2.3 Diameter of cell wall pores and hydrated

µmol (g root fresh wt)−1

(A) Transmembrane (B) Lipid linked (C) Protein attached

The most abundant membrane lipids are (i) phospholi-

2.4 SOLUTE TRANSPORT ACROSS

GS-X Ca2+ H+ Na+

ATP ATP ATP

Symport Antiport Uniport

Pumps Carriers Channel

Primary active transport Secondary Passive transport

(A) environmental signals including exposure to salt and low

gradients (White and Broadley, 2003; Martinoia et al., 2007;

υ (µmol K+ [g fresh wt]−1 h−1)

limitations of Michaelis-Menten kinetics and the presence

environmental conditions (Deane-Drummond and Chaffey,

0.4 0.4 SO42−

0.2 NH4+ 0.2

TABLE 2.9 Short-term P uptake parameters of soybean plants with different P

µmol K+ [(g fresh wt)−1 3 h−1]

the rate and selectivity of ion influx to root cells. As the

mic pH can affect ion uptake.

(Fig. 2.18), with tissue cation concentrations being more

2.5.3 Metabolic Activity

reactions show much greater temperature dependence with

uptake of ions from solutions of low concentration often

[µmol (g fw)−1 h−1]

(NH4)2SO4 (mM) K K K K MgCl2 

The interactions between selenate and sulphate are also

TABLE 2.18 K/Na selectivity of roots with or without Ca2 supply

1 mM CaCl2 5.9 15.0 20.9 15.4 10.7 26.1

NH4  55 43 22 120 0 23 33 5 59 120

5 NO3- + NH4+ External conc. Induced Induced

al., 1999; Buchner et al., 2004; Li et al., 2007; White and

conditions in particular, the conditions in the root environ-

Root NO3− concentration (mol g−1)

NO3− influx [nmol g−1 h−1]

of N supply (Lee and Rudge, 1986; Rawat et al.,

Although the transport systems induced by nutri-

TABLE 2.25 Relationship between the rate of K uptake Rhizosphere Cytoplasm

of nutrients and/or their metabolites from the shoot to the

Fe 7.8 4.8 96.8 2.60

COOH COOH COOH Rhizosphere Apoplasm Cytoplasm

Mugineic acid (MA)

H2C H C MnII FeIII

[µmol Fe equiv. (g root dry wt)−1 h−1]

graminaceous species. These differences are, as a rule,

TABLE 2.29 Rate of P uptake by various root zones of

Root zone (distance from

FIGURE 2.31 Schematic representation of anatomical changes along

The vacuoles of root cells also remove potentially

K Ca2 Ion concentration (mM)

O2 8.83 16.6 1.8 Potassium 23.3 24.5

N2 1.90 15.2 1.7 Calcium 9.1 9.5

Marschner (1995). Nitrate 18.1 0.0

osmometer. Temperature has a marked effect on the rate of

Root NO3− concentration (mol g−1)