Review

Uptake, allocation and signaling of

nitrate
Ya-Yun Wang, Po-Kai Hsu and Yi-Fang Tsay
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan

Plants need to acquire nitrogen (N) efficiently from the
soil for growth. Nitrate is one of the major N sources for
higher plants. Therefore, nitrate uptake and allocation
are key factors in efficient N utilization. Membrane-
bound transporters are required for nitrate uptake from
the soil and for the inter- and intracellular movement of
nitrate inside the plants. Four gene families, nitrate
transporter 1/peptide transporter (NRT1/PTR), NRT2,
chloride channel (CLC), and slow anion channel-
associated 1 homolog 3 (SLAC1/SLAH), are involved
in nitrate uptake, allocation, and storage in higher
plants. Recent studies of these transporters or channels
have provided new insights into the molecular mecha-
nisms of nitrate uptake and allocation. Interestingly,
several of these transporters also play versatile roles in
nitrate sensing, plant development, pathogen defense,
and/or stress response.
Nitrogen for plant growth
N, a building block of fundamental biological molecules,
such as nucleotides, amino acids, and proteins, is a key
nutrient for plant growth and development. Because N is a
major limiting factor in plant growth, considerable
amounts of N fertilizers are applied each year to increase
crop or fruit yields. However, excess use of N fertilizer has
caused pollution of rivers and oceans, leading to eutrophi-
cation and an increase in marine dead zones [1,2]. To
reduce the consumption of N fertilizer and increase N
utilization efficiency (NUE) in plants, an understanding
of how plants take up, store, and metabolize N has become
an urgent issue.
Most terrestrial plants absorb N from the soil. Both
inorganic and organic forms of N can be directly taken up
by plants. The concentrations of these two types of N in the
soil vary depending on soil type, temperature, and the
activities of microorganisms. Inorganic forms of N, such
as nitrate, ammonium, and urea, predominate in cropland,
and are often supplied in fertilizers; by contrast, organic
forms of N, such as amino acids, peptides, and proteins, are
dominant in the soils of boreal forests [3]. Being immobile
organisms, plants cannot choose their surrounding envir-
onments but only adapt to them. Therefore, they have
evolved several uptake and transport systems to optimize
and support their growth in response to rapidly changing
conditions. Using Arabidopsis (Arabidopsis thaliana) as a
model plant, several N uptake and transport systems have
been identified [4–11]. Nitrate is one of the major N sources
Corresponding author: Tsay, Y.-F. (yftsay@gate.sinica.edu.tw)
458 1360-1385/$ – see front matter ß 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.tplants.2012.04.006 Trends in Plant Science, August 2012, Vol. 17, No. 8
for higher plants. In this review, we focus on the physio-
logical functions of NRT1, NRT2, CLCa/b in the CLC
family, and SLAH3 in the uptake, allocation, and sensing
of nitrate (Figure 1).
Nitrate transporter and channels in higher plants
Chlorate resistant 1 (CHL1) was the first NRT1 transport-
er to be identified. It was identified as a nitrate uptake
mutant using chlorate selection [12]. The family to which
CHL1 belongs was named the NRT1/PTR family because
NRT1/PTR members in animals, fungi, and bacteria trans-
port dipeptides. In Arabidopsis, there are 53 NRT1/PTR
transporters. Of the 16 members characterized so far, some
transport nitrate and some transport dipeptides [13–25].
With the exception of CHL1 (AtNRT1.1) and MtNRT1.3,
which are dual-affinity nitrate transporters [26–28], most
of the NRT1 nitrate transporters characterized are low-
affinity nitrate transporters. Given that nitrate transport
activity has only been found in plant members of the PTR/
NRT1 family, it is likely that the nitrate transport activity
of this family evolved from an ancient dipeptide transport-
er. Most nitrate and peptide transporters characterized in
the NRT1/PTR family are proton-coupled transporters
[14,16,18,19,22,25,27].
In contrast to NRT1/PTR transporters, which have
diverse substrate specificity, all NRT2 transporters isolat-
ed from Aspergillus, Chlamydomonas, and higher plants
transport nitrate [5,9,29,30]. In Chlamydomonas and
higher plants, a membrane protein, NAR2, is required
for the nitrate transport activity of NRT2 transporters
(Figure 2) [31–36]. It is believed that the NRT2s are also
proton-coupled transporters. In Arabidopsis, there are
seven NRT2 transporters, four of which show nitrate-
related phenotypes when mutated [37–41].
The third type of nitrate transporters belongs to the
CLC family, some members of which function as anion
channels, and others as anion–proton exchangers [8]. In
Arabidopsis, there are seven CLC genes. CLCa and CLCb
function as proton–nitrate exchangers, and have a higher
selectivity for nitrate over chloride [42–44]. For CLCa, the
proline residue at 160, which is also found at the corre-
sponding position in CLCb, is important for nitrate selec-
tivity [45–47].
Studies of an Arabidopsis mutant defective in stomatal
closure has led to the identification of the anion channel
SLAC1 [48,49]. SLAC1 contains ten transmembrane
domains and shares sequence similarity with bacterial
and fungal dicarboxylate transporters. In Arabidopsis,
 Review Trends in Plant Science August 2012, Vol. 17, No. 8

(a) Embryo cell

Vacuole

NRT2.7

NR

Se
ed
T1
NO3– H+ (?)

:6
Mesophyll cell
Vacuole
H+

CLCa,b
:7
T1 e NO3–
NR b iliz
mo
Re
Guard cell
NRT1.1

e
.4 or
RT1 e st
N l
tio SLAH3
Pe
NRT1.5
Xylem

NRT2.1
NRT2.2
HATS
NRT1.8 Uptake NRT2.4
NRT1.9 NRT1.1(CHL1)
LATS
NRT1.2
Efflu
x
NAXT1

(b) Xylem
Pericycle/
Epidermis Cortex Endodermis Parenchyma
NO 3–
NO3–
NO3–
NO3– NO 3–
NO3– NO3– NO3–
NO3 –

NO3– NO 3–

NO3– NO3–
Casparian band

Key: NRT1.1(CHL1) NRT1.2 NAXT1 NRT1.8

NRT2.1 NRT2.4 NRT1.5

TRENDS in Plant Science

Figure 1. Physiological functions of Arabidopsis nitrate transporters. (a) Contribution of nitrate transporter 1 (NRT1), NRT2, chloride channel (CLC) a/b, and slow anion
channel-associated 1 homolog 3 (SLAH3) to different steps of nitrate uptake and allocation. Nitrate is taken up by roots from soils and transported to shoots and seeds for
storage and/or further assimilation. Abbreviations: HATS, high-affinity transport system; LATS, low-affinity transport system. (b) Detailed illustration of nitrate uptake and
movement in roots. NRT1.1 (CHL1), NRT1.2, NRT2.1, NRT2.2, and NRT2.4 are involved in nitrate uptake from soils. Nitrate excretion transporter 1 (NAXT1), a transporter in
the NRT1 family, mediates nitrate efflux under acid load. NRT1.5, NRT1.8, and NRT1.9 play a role in regulating root-to-shoot xylem transport of nitrate. The illustration does
not mean that these transporters have a polarized distribution in the root cells.

there are five SLAC genes: SLAC1 and SLAH1–SLAH4
(SLAC1 homologs). When expressed in oocytes, SLAC1 and
SLAH3 show nitrate transport activity, although kinases
need to be co-injected to activate their channel activity
[50,51].
Acquisition of nitrate from soil
Nitrate concentrations in the soil can fluctuate by more
than four orders of magnitude [52,53]. To survive, plants
have evolved two nitrate uptake systems, the high-affinity
transport system (HATS), and the low-affinity transport
459
 Review Trends in Plant Science August 2012, Vol. 17, No. 8

NO3–
high-affinity nitrate uptake [37,40,41], with NRT2.4 hav-
Cytosol Vacuole ing a very high-affinity range [41]. The relative contribu-
NAXT1 NO3–
tion of these transporters to nitrate uptake is dependent on
H+ (?) the developmental stages of the root and the N status of the

CL
H+

Ca
NO3– plant.

b,
NRT2.7 The Km values of plants with an HATS are in the micro-
NRT1.5
molar range. Consistently, the Km value of HvNRT2.1 and
H+ CIPK23 NO3– H+(?) the high-affinity Km value of CHL1 (AtNRT1.1) measured
CPK21
when expressed in Xenopus oocytes have been found to be
PO4– PO4– approximately 30 mM and approximately 50 mM, respec-

NAR2
NRT1 NRT2 SLAH3
– +
NO3– H+ NO3 H NO3–
TRENDS in Plant Science
Figure 2. Directions of nitrate movement mediated by nitrate transporter 1 (NRT1),
NRT2, chloride channel (CLC) a,b, and slow anion channel-associated 1 homolog 3
(SLAH3) in plasma membrane or tonoplast. Most NRT1 and NRT2 transporters
characterized are H+-coupled symporters. NRT1.1 (CHL1), a dual-affinity nitrate
transporter, is phosphorylated by calcineurin B-like-interacting protein kinase 23
(CIPK23) to change the affinity. NAR2 is required for NRT2 nitrate transport. CLCa,b
localized in the tonoplast are H+/NO3 antiporters. Anion channel SLAH3 mediates
nitrate release from guard cells and its function is regulated by CPK21.
Abbreviations: CPK21, calcium-dependant protein kinase 21; NAXT1, nitrate
excretion transporter 1.
system (LATS). Thermodynamic calculations as well as the
association of both the HATS and the LATS with mem-
brane depolarization suggest that both systems require
energy potentially provided by proton gradients [54–56]. In
Arabidopsis, CHL1 is a dual-affinity nitrate transporter
involved in both high- and low-affinity nitrate uptake
[26,27]. In addition to CHL1, NRT1.2, in the NRT1 family,
also participates in low-affinity nitrate uptake [20]. In the
NRT2 family, NRT2.1, NRT2.2, and NRT2.4 participate in
tively [27,57]. The Km value of AtNRT1.2, as well as the
low-affinity Km value of CHL1 (AtNRT1.1), are both approx-
imately 5 mM when expressed in Xenopus oocytes
[20,27,58]. Consistent with these measurements in oocytes,
some plant nitrate uptake studies showed that the LATS
exhibits a saturable response with a Km value in the mM
range [26,59,60]. However, in other studies, the plant LATS
showed a linear response to concentration [61–63]. Accord-
ing to thermodynamic calculations, it is unlikely that plant
LATS is mediated by the passive transport of a channel. The
linear response might need to be reevaluated with wider
ranges of concentrations. Nevertheless, it might be possible
that a channel facilitates nitrate uptake when the mem-
brane potential is depolarized and/or the cytosolic nitrate
concentration is low.
Nitrate uptake is regulated at both the transcriptional
and post-transcriptional level. As shown in Table 1, ex-
pression of CHL1 (NRT1.1), NRT1.2, NRT2.1, NRT2.2,
and/or NRT2.4 is regulated at the transcriptional level
by nitrate, nitrite, ammonium, glutamine, N starvation,
light, sucrose, diurnal rhythm, and/or pH. A strong corre-
lation between the transcript abundance of these nitrate
transporter genes and nitrate uptake activities suggests

Table 1. Summary of the physiological functions and regulations of identified nitrate transporters in Arabidopsis thaliana
Gene Function Nitrate response N starvation Other regulations Refs
CHL1 (NRT1.1) Nitrate sensing Induction Repression Nitrite (long-term) and high [12,27,54,58,66,
pH repression 70,103–108]
High- and low-affinity nitrate uptake Auxin, light, sugar, and nitrite
(short-term) induction
NRT1.2 Low-affinity nitrate uptake Constitutive Not known Not known [27,109]
NRT1.3 Not known Induction (shoot) Not known Light induction [108,109]
NRT1.4 Leaf nitrate homeostasis Constitutive Not known Not known [18,109]
NRT1.5 Root xylem loading Induction Not known High pH and potassium [14,108]
limitation repression
Sugar induction
NRT1.6 Delivery of nitrate to developing embryos Not known Induction Not known [16]
NRT1.7 Nitrate remobilization from old to Not known Induction Sucrose induction [25]
young leaves
NRT1.8 Xylem unloading and cadmium resistance Induction Not known Cadmium induction [13]
NRT1.9 Nitrate loading into root phloem Constitutive Not known Not known [22]
NAXT1 Root nitrate efflux Not known Not known Acidic pH induced at [76]
protein level
NRT2.1 High-affinity nitrate uptake Induction Induction Ammonium and [37,40,65,66,
glutamine repression 105,106,108–112]
Light and sugar induction
NRT2.2 High-affinity nitrate uptake Induction Not known Not known [37,40,109,112]
NRT2.4 High-affinity nitrate uptake at low Repression Induction Ammonium repression [41,108,109]
nitrate concentration
Light induction
NRT2.7 Nitrate storage in mature embryos Constitutive Not known Not known [39,109]
CLCa Nitrate accumulation in vacuoles Induction Not known Not known [43,44]
CLCb Nitrate accumulation in vacuoles Not known Not known Not known [42]

460
Review
that transcriptional regulation plays a key role in modu-
lating nitrate uptake activities. Transcript abundance of
CHL1 (NRT1.1) and/or NRT2.1 has been particularly
linked to response to nitrate induction, N-starvation upre-
gulation, ammonium repression, and/or diurnal changes
[36,40,64–66].
Studies of 35S–NRT2.1 transgenic Arabidopsis and
Nicotiana have revealed post-transcriptional repression
of nitrate uptake in response to ammonium or dark treat-
ments [36,67]. In wild type, both the AtNRT2.1 mRNA
level and the HATS are downregulated by prolonged dark.
In the Arabidopsis 35S–NRT2.1 plant, the mRNA level
remained unchanged in response to prolonged dark, but
the HATS and NRT2.1 protein levels were reduced. These
results indicate that, in addition to transcriptional control,
protein stability also provides a path through which the
HATS is repressed by prolonged (24-h) dark treatments
[36]. Another example of post-transcriptional regulation
was revealed in a study of NAR2. In Arabidopsis and
Chlamydomonas, an oocyte functional assay as well as
mutant phenotypes indicated that the two-transmem-
brane-domain protein NAR2 is required for the nitrate
transport activity of the NRT2 transporter. In the Arabi-
dopsis nar2.1 mutant, the disappearance of NRT2.1
protein in the membrane fraction suggested that
NAR2.1 is required for the plasma membrane targeting,
and/or the protein stability, of NRT2.1 [68].
Protein phosphorylation provides another regulatory
route for nitrate uptake. CHL1 (NRT1.1), expressed in
root epidermis, cortex, and endodermis, is a dual-affinity
nitrate transporter mediating both the HATS and the
LATS [26,27,58]. The switch between the two affinities
is controlled by phosphorylation at the T101 residue be-
tween the second and third transmembrane domains [69],
and this phosphorylation is regulated by the calcineurin B-
like-interacting protein kinase CIPK23 [70]. Dual-affinity
transport activity is not unique to CHL1, as it is also
exhibited by the potassium transporter KUP and the
nitrate transporter MtNRT1.3 [28,71,72]. Furthermore,
some of the transporters characterized to date might not
have been examined with wide enough ranges of substrate
concentrations to eliminate completely the possibility that
they are dual-affinity transporters. It is not known wheth-
er there are two nitrate binding sites with different affini-
ties in CHL1, or one binding site for both affinities that is
altered by conformational change. The crystal structure of
PepTso, a prokaryotic homolog of CHL1, revealed two
hydrophilic cavities, an extracellular cavity and a central
cavity [73]. The peptide binding site is proposed to be in the
central cavity. The leucine/sodium symporter LeuT, a
prokaryotic ortholog of neurotransmitter/sodium sympor-
ters (NSSs), also contains two hydrophilic cavities, and it
has been reported that it may have two leucine binding
sites, with one in each cavity [74]. It will be interesting to
determine whether there are also two substrate binding
sites in CHL1, and whether or how phosphorylation in the
cytosolic loop affects the accessibility and/or affinity of
these binding sites.
Involvement of multiple transporters in nitrate uptake
together with multiple routes of regulation would mean
that plants have a sophisticated uptake system fine-tuned
Trends in Plant Science August 2012, Vol. 17, No. 8
to optimize nitrate uptake capacity in response to internal
demand and external changes.
Nitrate efflux
Net root nitrate uptake is the balance between nitrate
influx (uptake) and efflux. Although the physiological role
of nitrate efflux remains unclear [75,76], it is known that it
increases upon external environment and other stresses
[77,78]. Using a functional biochemical approach, nitrate
excretion transporter 1 (NAXT1) was identified by mass
spectrometry in the plasma membrane of Arabidopsis
suspension cells (Figures 1 and 2) [76]. Interestingly,
NAXT1, a member of the NRT1/PTR family, is mainly
expressed in the cortex of mature roots. Under normal
conditions, nitrate efflux activity in wild type and naxt1
mutants is similar. However, acidification of the cytosol by
acid load stimulated nitrate efflux in wild type but not in
naxt1 mutants, indicating that NAXT1 is responsible for
nitrate excretion induced by lower pH in the cytosol [76].
The appearance of phenotypes of naxt1 under acid load
conditions is consistent with the fact that NAXT1 belongs
to the proton-coupled symporter family. However, direct
evidence for the proton-coupling mechanism of NAXT1 is
still missing. Similar to NAXT1, NRT1.5, which is respon-
sible for loading nitrate into the xylem, also mediates
nitrate efflux (see next section) [14]. It is generally believed
that nitrate influx is an active process, with energy
obtained from proton coupling, whereas nitrate efflux is
a passive process, which is more likely to be mediated by a
channel. Therefore, it was surprising that two transporters
in the proton-coupled transporter family, NAXT1 and
NRT1.5, were found to be involved in nitrate efflux. The
efflux activity of NRT1.5 is pH dependent, suggesting that
efflux is also mediated by a proton-coupled mechanism. It
is worth noting that SLAH3 (a homolog of SLAC1), when
coexpressed with a calcium-dependent protein kinase
(CPK21) in Xenopus oocytes, exhibits a nitrate-induced
anion current [50]. SLAH3 shows strong selectivity for
nitrate over chloride. Given that no nitrate efflux differ-
ence can be detected in the naxt1 mutant under normal
conditions, it is possible that an unknown channel or a
homolog of SLAH3 is responsible for root nitrate efflux
under normal conditions. Moreover, the influence of nitrate
excretion on plant growth remains to be analyzed further.
Translocation from root to shoot
Once inside plant cells, nitrate can be stored in vacuoles or
assimilated to ammonium for amino acid synthesis. Under
many conditions, a significant proportion of nitrate assim-
ilation takes place in shoots because the reducing power
required for the assimilation processes comes from photo-
synthesis. For root-to-shoot nitrate translocation, the first
step is the loading of nitrate into xylem vessels. To date,
only one nitrate transporter, NRT1.5, has been identified
to mediate this process (Figures 1 and 3).
NRT1.5, a low-affinity nitrate transporter located in the
plasma membrane, is expressed in the two pericycle cells
adjacent to the protoxylem (the xylem-pole pericycle) in
Arabidopsis roots (Figures 2 and 3) [14]. In nrt1.5 mutants,
less nitrate is transported to the shoots and the nitrate
content in xylem sap is reduced [13,14], suggesting that
461
Review
Root-to shoot xylem
transport of nitrate
Co
Pe
Xy
En Ph
Downward phloem
transport of nitrate
Nitrate removal
from xylem
TRENDS in Plant Science
Figure 3. Regulation of root-to-shoot nitrate transport. Nitrate transporter 1.5
(NRT1.5), expressed in the xylem pole pericycle, mediates xylem-loading of nitrate
(red arrow) and facilitates root-to-shoot xylem transport of nitrate (blue arrow).
NRT1.9, expressed in companion cells of phloem, mediates phloem loading (dark-
purple arrow) for downward phloem transport of nitrate (dark blue arrow), which
may facilitate xylem-to-phloem transfer of nitrate (broken purple arrow), and then
reduce root-to-shoot nitrate transport. NRT1.8, expressed in xylem parenchyma,
mediates nitrate removal from xylem (green arrow), and could also reduce root-to-
shoot nitrate transport. Abbreviations: Co, cortex; En, endodermis; Pe, pericycle;
Ph, phloem; Xy, xylem.
NRT1.5 is responsible for exporting nitrate out of pericycle
cells for xylem loading. Interestingly, in the mutants, the
amount of potassium transported to the shoot is also
reduced in a nitrate-dependent manner [14], suggesting
that there is homeostatic balance between the major cat-
ion, potassium, and the anion, nitrate, in the xylem stream.
NRT1.5 is a bidirectional (influx–efflux) nitrate transport-
er. Influx activity is higher when the external solution is
more acidic (pH 5.5), whereas efflux activity is more obvi-
ous at neutral pH values (pH 7.4). Given that the pH value
of xylem sap often ranges from 5.7 to 6.2 [79–81], further
study is required to determine how the two directions of
nitrate movement are controlled. Root-to-shoot nitrate
transport is not completely abolished in the nrt1.5 mutant,
and when tissue nitrate content is low, no nitrate translo-
cation defect can be detected [14]; therefore, there must be
other xylem loading mechanism(s) that are more active
under low nitrate conditions.
Two NRT1 transporters, NRT1.8 and NRT1.9, are in-
volved in regulating root-to-shoot nitrate translocation
(Figure 1). NRT1.8, a low-affinity nitrate transporter lo-
cated in the plasma membrane, is expressed dominantly in
the xylem parenchyma cells of roots (Figure 3) [13]. nrt1.8
mutants show increased nitrate content in xylem sap and
increased root-to-shoot nitrate translocation, suggesting
that NRT1.8 functions in removing nitrate from the xylem
sap back into the root cells [13]. Studies of NRT1.8 have
revealed an interesting interplay between nitrate and
cadmium. In wild type, expression of NRT1.8 is upregu-
lated by cadmium, and cadmium treatment reduces the
amount of nitrate translocated to the shoot. The nrt1.8
mutant shows a nitrate-dependent cadmium-sensitive
phenotype and, compared with the wild type, an increased
amount of cadmium is transported to the shoot. Therefore,
although the detailed mechanism remains to be deter-
mined, it seems that the function of NRT1.8 in removing
462
Trends in Plant Science August 2012, Vol. 17, No. 8
nitrate from xylem sap also allows Cd2+ to stay in the roots,
and consequently enhances Cd2+ tolerance [13].
In a similar manner to nrt1.8 mutants, nrt1.9 mutants
also show enhanced root-to-shoot nitrate transport [22]
(Figure 1). However, unlike NRT1.8, which is expressed
in xylem parenchyma cells, NRT1.9 is expressed in the
companion cells of the phloem (Figure 3), indicating that
both NRT1.8 and NRT1.9 are negative regulators of root-
to-shoot nitrate transport but through different mecha-
nisms. In nrt1.9 mutants, downward transport of nitrate
through the phloem is reduced, resulting in increased
upward (root-to-shoot xylem) transport of nitrate [22].
Moreover, nrt1.9 mutants show enhanced plant growth
under high nitrate conditions. These results suggest that
NRT1.9 mediates the downward phloem transport of ni-
trate in roots and that, under high nitrate conditions,
phloem transfer of nitrate in the root serves as an ‘overflow
route’ to prevent excess amounts of nitrate being trans-
ported to the shoot. Under laboratory growth conditions,
nrt1.9 mutants gain a growth advantage under high ni-
trate conditions, but the lack of this additional regulatory
path in nrt1.9 mutants may result in a loss of flexibility to
adapt to certain environmental changes in the wild.
Studies of NRT1.5, NRT1.8, and NRT1.9 indicate that
the root-to-shoot xylem transport of nitrate is a key step in
regulating nitrate distribution and so needs to be fine
tuned by several accessory pathways. Root-to-shoot nitrate
transport is also crucial for the homeostatic interplay
between nitrate and other cations.
Allocation in vegetative tissues
After transportation to leaves, nitrate can undergo assimi-
lation in cytosol or storage in vacuoles. NRT1.4, a low-
affinity nitrate transporter, plays a role in regulating leaf
nitrate homeostasis and leaf development (Figure 1) [18].
NRT1.4 is dominantly expressed in the leaf petioles. The
nitrate content of the petiole is reduced but the nitrate
content in the leaf lamina is slightly increased in nrt1.4
mutants, indicating that NRT1.4 regulates nitrate distri-
bution in leaves. Interestingly, the leaves of nrt1.4 mutants
are wider than those of the wild type, suggesting that the
nitrate distribution in leaves influences leaf expansion.
These studies indicate that the petiole, a nitrate storage
site, serves as a checkpoint for regulating nitrate distribu-
tion in leaves. Interestingly, in some crop species, farmers
use petiole nitrate content to monitor the N fertilizer
demands of the crops [82,83]. Moreover, NRT1.4 expressed
in the petiole could affect lamina nitrate content and
lamina growth, suggesting that there are multiple layers
of interplay between the lamina and petiole for nitrate
distribution and assimilation. For example, it would be
interesting to find out when and how N demand in the
lamina sends a signal to the petiole to trigger the remobi-
lization of stored nitrate.
It is well known that sucrose can be allocated from
source (mature) leaves to sink (young) leaves to support
the growth of sink tissues with low levels of photosynthesis
activity. A study of NRT1.7 has shown that nitrate can also
be allocated from older leaves to younger leaves (Figure 1)
[25]. NRT1.7 is expressed in the phloem of minor veins in
older leaves (Figure 4). In nrt1.7 mutants, more nitrate
 Review
Younger leaves Older leaves
O3–
NO
NO3– Xy
Source-to-sink
remobilization of nitrate
Ph
TRENDS in Plant Science
Figure 4. Function of nitrate transporter 1.7 (NRT1.7) in nitrate remobilization from
older to younger leaves. Root-to-shoot xylem transport of nitrate (red arrows) is
driven by transpiration, so less nitrate is transported to the smaller younger leaves
via the xylem. NRT1.7 (yellow circles), expressed in the phloem of the minor veins
in older leaves, mediates phloem-loading of nitrate in older leaves (short blue
arrows) and facilitates remobilization of excess nitrate stored in older leaves into
younger leaves (long blue arrow). Abbreviations: Ph, phloem; Xy, xylem.
accumulated in older leaves, less 15NO3– dropped on the
older leaves could be transported to younger leaves, and
less nitrate was detected in the phloem sap of the older
leaves. These findings indicate that phloem loading medi-
ated by NRT1.7 in the minor veins of older leaves could
remove excess nitrate from older leaves to feed younger
leaves. In addition to NRT1.7, NRT2.4 also participates in
phloem nitrate transport in shoots, particularly under N
starvation conditions [41]. NRT2.4 is expressed in the
phloem parenchyma of leaves, and the expression is highly
induced by N starvation. Consistent with expression pat-
terns, the nitrate content in the phloem exudates of mature
leaves in nrt2.4 mutants is decreased under N starvation
conditions but not under normal conditions, suggesting a
special role for NRT2.4 in phloem-mediated nitrate remo-
bilization. [41]. Intriguing questions are raised, particular-
ly relating to the temporal and spatial differences between
NRT1.7 and NRT2.4 in their contribution to nitrate remo-
bilization under normal and starvation conditions.
For some time, it was generally believed that nitrate
could only be transported via the xylem, whereas organic
N, such as amino acids, could be transported via both the
xylem and phloem. Therefore, it was surprising to find that
phloem nitrate transport mediated by NRT1.7 and NRT1.9
plays a crucial role in controlling nitrate distribution.
Based on this new information, the understanding of ni-
trate allocation has been modified: it is now thought that
root-to-shoot xylem transport driven by transpiration is
the primary route of long-distance nitrate transport and
that phloem nitrate transport driven by osmotic gradients
provides a secondary route by which to modulate local
nitrate redistribution.
NRT1.7 expression is diurnally regulated, with levels
peaking at dusk. This is the opposite situation to that of the
major nitrate reductase gene NIA2, suggesting that nitrate
in older leaves is assimilated during the light period and
exported during the dark period. Expression of NRT1.7 is
upregulated by N starvation and, under N starvation
Trends in Plant Science August 2012, Vol. 17, No. 8
conditions, nrt1.7 mutants show growth retardation, sug-
gesting that remobilization of nitrate among leaves is
important to sustain plant growth under such conditions
[25]. Because excess nitrate is stored in the vacuoles to
await export from the leaf, nitrate has to be transported
across the tonoplast and the plasma membrane of the
mesophyll cells before it can be loaded into the phloem.
How N starvation and/or N demand from the younger
leaves could coordinately regulate these steps is an inter-
esting question.
The genes responsible for exporting nitrate out of
vacuoles have not yet been identified. However, the genes
responsible for transporting nitrate into vacuoles are well
documented. CLCa and CLCb, proton–nitrate exchangers
located in the tonoplast, mediate nitrate import into the
vacuole (Figures 1 and 2) [42,43]. CLCa expressed in meso-
phyll cells accounts for up to 50% of nitrate storage [44]. The
expression level of CLCb is much lower than that of CLCa. In
contrast to the clca mutant, the clcb mutant shows no
difference in nitrate content compared with the wild type
[42]. Both CLCa and CLCb are diurnally regulated and
upregulated by nitrate. CLCa activity can be inhibited by
the binding of ATP to the cystathionine-synthetase (CBS)
domains at its cytosolic C-terminus [84]. This suggests that
nitrate storage activity is regulated by the energy and
photosynthesis status of the mesophyll cells, and that
nitrate storage activity consequently affects nitrate assimi-
lation rate as this is determined by nitrate reductase activity
and cytosolic nitrate concentration.
Nitrate in guard cells
Nitrate is one of the major anionic osmotica in higher
plants. Stomatal closure and opening are driven by
changes in turgor pressure, and several nitrate transpor-
ters and channels are associated with stomatal pheno-
types. CHL1 is expressed not only in roots, but also in
the guard cells in leaves (Figure 1) [85]. In the presence of
nitrate, CHL1 expressed in guard cells promotes stomatal
opening. In chl1 mutants, less nitrate is accumulated in
guard cells, and stomatal opening and transpiration rates
are reduced in the presence of nitrate. Therefore, chl1
mutants show more drought tolerance than does the wild
type in the presence of nitrate [85]. Interestingly, the
transcription factor NIN-like protein 7 (NLP7), a positive
regulator of nitrate-induced expression of N-related genes,
including CHL1, also shows drought tolerance when mu-
tated [86]. These results further strengthen a linkage
between nitrate and water loss.
Anion efflux is a key player in stomatal closure. SLAC1,
identified by stomatal closure mutants, exhibits similar
permeability for nitrate and chloride [51]. In contrast to
SLAC1, SLAH3 shows a strong preference for nitrate over
chloride (Figures 1 and 2) [50]. Even though SLAH3 shows
similar anion selectivity to the S-type anion current found
in guard cell protoplasts, no stomatal phenotype has been
seen in slah3 mutants under the conditions tested [50].
These findings suggest that, under normal conditions,
SLAC1 is more dominant in regulating stomatal closure.
In addition to CPK, high external nitrate is required to
activate SLAH3 with K0.5 of 8.3 mM [50]. Therefore, under
nitrate-rich conditions, the nitrate-preferred channel can
463
 Review
F changing the phosphorylation status at the T101 residue,
Nitrate supply for early
em
embryo development
Nitrate storage in
mature embryos
TRENDS in Plant Science
Figure 5. Nitrate transport to embryos. Nitrate transporter 1.6 (NRT1.6), expressed
in the funiculus (F), provides nitrate for early embryo development (orange arrow).
NRT2.7, located in the tonoplast, mediates nitrate storage in mature embryos (blue
arrow). The GUS (blue) staining and the GFP (green) signals in this photo
represents NRT1.6 and NRT2.7 expression patterns, respectively. This figure was
produced by combining two micrographs reproduced, with permission, from
[20,44].
be activated to substitute for the nonselective one in regu-
lating stomatal closure.
Movement in reproductive tissues
In addition to organic N, nitrate also accumulates in seeds
and affects seed dormancy [87]. NRT1.6, a plasma-mem-
brane localized, low-affinity nitrate transporter, is only
expressed in the vascular bundles of the siliques and the
funiculi, suggesting that it is involved in delivering nitrate
to developing seeds (Figures 1 and 5). Indeed, in nrt1.6
mutants, the amount of nitrate in mature seeds is reduced
and the seed abortion rate is increased [16]. The abnormal
embryos are caused by the collapse of the suspensor cells at
the one- or two-cell stage, indicating that nitrate is impor-
tant for early embryo development.
In the NRT2 family, NRT2.7 is expressed in the mature
seeds and located in the tonoplast (Figures 2 and 5) [39].
NRT2.7 alone without NAR2 can mediate nitrate uptake in
Xenopus oocytes and less nitrate is detected in nrt2.7 seeds,
indicating that NRT2.7 plays a role in nitrate accumula-
tion in seeds. If, as expected, NRT2.7 functions as a proton-
coupled nitrate transporter, one would expect that NRT2.7
located in the tonoplast is involved in transporting nitrate
out of the vacuole. However, reduced seed nitrate content
suggests that NRT2.7 is involved in importing nitrate into
the vacuole (Figure 2). The magnitude of the reduction in
seed nitrate content in the mutants is ecotype dependent
[39], indicating that different ecotypes might adapt differ-
ent mechanisms for nitrate storage in seeds.
Versatile roles of nitrate transceptor CHL1
In addition to being a transporter involved in nitrate
uptake, CHL1 also functions as a nitrate sensor, regulating
a transcriptional response called the primary nitrate re-
sponse [70,88]. In the primary response, several nitrate-
related genes, including NRT1.1, NRT2.1, the nitrate re-
ductase-encoding genes NIA1 and NIA2, and the nitrite
reductase-encoding gene NIR, are induced by nitrate with-
in 10 min. A study of the uptake- and sensing-decoupled
mutant chl1-9 (P492L) showed that nitrate does not need
to be transported across the membrane when CHL1 exerts
464
Trends in Plant Science August 2012, Vol. 17, No. 8
its function as a nitrate sensor to trigger the signal [70]. By
CHL1 can sense the concentration changes in the external
environment, leading to different levels of primary re-
sponse. At a low nitrate concentration, T101 is phosphory-
lated by CIPK23, and phosphorylated CHL1 triggers a low-
level of primary response [70].
CHL1 also plays a role in regulating root architecture.
The growth of the primary root in wild type is inhibited by
exogenous L-glutamate (Glu). This inhibition effect is an-
tagonized by nitrate. However, the antagonism by nitrate
is lost in chl1 mutants, suggesting that CHL1 senses the
presence of nitrate and mediates the effect [89].
In addition to primary root growth, CHL1 (NRT1.1) also
participates in promoting lateral root elongation in nitrate-
rich patches of the external medium [90]. A study has
shown that, in the absence of nitrate, CHL1 (NRT1.1)
facilitates the uptake of the phytohormone auxin, whereas
in the presence of nitrate, CHL1-dependent auxin trans-
port is inhibited [91]. This study also showed that this
nitrate-regulated CHL1-dependent auxin transport is re-
sponsible for nitrate-promoted lateral root elongation. In
addition, ethylene is involved in repressing lateral root
development under high nitrate conditions, and this re-
pression is alleviated in ethylene-insensitive mutants
(etr1-3 and ein2-1) and in chl1 mutants [92]. The expres-
sion of CHL1 is also affected by ethylene signaling, sug-
gesting that CHL1 (NRT1.1) is also involved in the
ethylene-mediated inhibition of lateral root development.
As well as having nitrate-related functions, several
reports have shown that CHL1 also has nitrate-indepen-
dent functions [93–96]. The chl1 mutant has an acidic
intracellular pH and reduced primary root growth in the
absence of nitrate [95,96]. Mutation of CHL1 (NRT1.1) also
enhances ammonium and/or low pH tolerance in Arabi-
dopsis and real-time PCR analysis suggests that this
tolerance results from the upregulation of aliphatic gluco-
sinolate biosynthesis through a nitrate-independent signal
from CHL1 [93].
Effect of NRT2.1 on lateral root development and
pathogen defense
The high-affinity uptake complex NRT2.1–NAR2.1 also
participates in regulating lateral root development. Under
nitrate-limiting conditions, both genes are positive regu-
lators of lateral root initiation even in nitrate-free condi-
tions, suggesting that the regulation of lateral root growth
by NRT2.1 and NAR2.1 is independent of their uptake
function [33,97]. However, under high sucrose and low
nitrate conditions, NRT2.1 and NAR2.1 function as repres-
sors [33,98]. These results indicate that the NRT2.1–
NAR2.1 complex has dual effects on lateral root develop-
ment. Another surprising phenotype seen in nrt2.1
mutants is reduced susceptibility to the bacterial pathogen
Pseudomonas syringae pv tomato DC3000 (Pst) [99]. This
phenotype probably has reduced sensitivity to the bacterial
phytotoxin coronatine because no difference could be found
between the wild type and nrt2.1 mutant when challenged
with a P. syringae strain defective in coronatine synthesis.
The multiple effects of NRT1.1 and/or NRT2.1 on nitrate
signaling, root development, and pathogen defense suggest
Review
that, in addition to being a nutrient source, nitrate is a
signaling molecule coordinating multiple cellular process-
es. A challenge for future research will be to find out
whether these nitrate transporters are directly or indirect-
ly involved in these responses, and to find out the details of
the underling mechanism(s) and potential cross talk
among these responses.
Concluding remarks
Since the identification of the first member of the NRT1/
PTR family in 1993, considerable effort has been put into
characterizing the transport activities and in vivo func-
tions of other members in this family. Although some
progress has been made, the information available is still
just the tip of the iceberg. For example, there are at least 80
genes in the rice (Oryza sativa) NRT1 family [5,100] but to
date only two of them have been functionally characterized
[101,102]. Even in Arabidopsis, less than one third of
NRT1 and NRT2 genes have been characterized. A big
question that plant scientists have is why higher plants
need so many nitrate transporters. Considering that ni-
trate is a major N source and plants are multicellular
organisms with specialized cells and tissues, it is not
surprising that several nitrate transporters are required
to optimize plant growth in response to the external envi-
ronment. However, how those genes function and coordi-
nate external stimuli with internal growth requires more
study. As more details come to light, it might be possible to
build a holistic picture of nitrate usage in plants; this
knowledge could then be used to improve crop yield in
unfavorable environments or harsh climatic regions.
Conflict of interest
The authors declare that they have no conflicts of interest relevant to this
paper.
Acknowledgments
This work was supported by grants from the National Science Council
(NSC 100-2321-B-001-013 and NSC 100-2311-B-001-004-MY3) and the
Academia Sinica Postdoctoral Fellows Program.
References
1 Diaz, R.J. and Rosenberg, R. (2008) Spreading dead zones and
consequences for marine ecosystems. Science 321, 926–929
2 Canfield, D.E. et al. (2010) The evolution and future of earth’s nitrogen
cycle. Science 330, 192–196
3 Näsholm, T. et al. (2009) Uptake of organic nitrogen by plants. New
Phytol. 182, 31–48
4 Tegeder, M. and Rentsch, D. (2010) Uptake and partitioning of amino
acids and peptides. Mol. Plant 3, 997–1011
5 Tsay, Y.F. et al. (2007) Nitrate transporters and peptide transporters.
FEBS Lett. 581, 2290–2300
6 Kojima, S. et al. (2006) Molecular mechanisms of urea transport in
plants. J. Membr. Biol. 212, 83–91
7 Ludewig, U. et al. (2007) Molecular mechanisms of ammonium
transport and accumulation in plants. FEBS Lett. 581, 2301–2308
8 Barbier-Brygoo, H. et al. (2011) Anion channels/transporters in
plants: from molecular bases to regulatory networks. Annu. Rev.
Plant Biol. 62, 25–51
9 Dechorgnat, J. et al. (2011) From the soil to the seeds: the long journey
of nitrate in plants. J Exp. Bot. 62, 1349–1359
10 Tsay, Y-F. et al. (2011) Integration of nitrogen and potassium
signaling. Annu. Rev. Plant Biol. 62, 207–226
11 Ho, C-H. and Tsay, Y-F. (2011) Nitrate, ammonium, and potassium
sensing and signaling. Curr. Opin. Plant Biol. 13, 604–610
Trends in Plant Science August 2012, Vol. 17, No. 8
12 Tsay, Y.F. et al. (1993) The herbicide sensitivity gene CHL1 of
Arabidopsis encodes a nitrate-inducible nitrate transporter. Cell
72, 705–713
13 Li, J.Y. et al. (2010) The Arabidopsis nitrate transporter NRT1.8
functions in nitrate removal from the xylem sap and mediates
cadmium tolerance. Plant Cell 22, 1633–1646
14 Lin, S.H. et al. (2008) Mutation of the Arabidopsis NRT1.5 nitrate
transporter causes defective root-to-shoot nitrate transport. Plant
Cell 20, 2514–2528
15 Komarova, N.Y. et al. (2008) AtPTR1 and AtPTR5 transport
dipeptides in planta. Plant Physiol. 148, 856–869
16 Almagro, A. et al. (2008) Characterization of the Arabidopsis nitrate
transporter NRT1.6 reveals a role of nitrate in early embryo
development. Plant Cell 20, 3289–3299
17 Dietrich, D. et al. (2004) AtPTR1, a plasma membrane peptide
transporter expressed during seed germination and in vascular
tissue of Arabidopsis. Plant J. 40, 488–499
18 Chiu, C.C. et al. (2004) Mutation of a nitrate transporter, AtNRT1:4,
results in a reduced petiole nitrate content and altered leaf
development. Plant Cell Physiol. 45, 1139–1148
19 Chiang, C.S. et al. (2004) Mechanisms and functional properties of two
peptide transporters, AtPTR2 and fPTR2. J. Biol. Chem. 279, 30150–
30157
20 Huang, N.C. et al. (1999) Cloning and functional characterization of
an Arabidopsis nitrate transporter gene that encodes a constitutive
component of low-affinity uptake. Plant Cell 11, 1381–1392
21 Rentsch, D. et al. (1995) NTR1 encodes a high affinity oligopeptide
transporter in Arabidopsis. FEBS Lett. 370, 264–268
22 Wang, Y.Y. and Tsay, Y.F. (2011) Arabidopsis nitrate transporter
NRT1.9 is important in phloem nitrate transport. Plant Cell 23,
1945–1957
23 Karim, S. et al. (2005) Structural and functional characterization of
AtPTR3, a stress-induced peptide transporter of Arabidopsis. J. Mol.
Model 11, 226–236
24 Song, W. et al. (1996) Cloning of a second Arabidopsis peptide
transport gene. Plant Physiol. 110, 171–178
25 Fan, S.C. et al. (2009) The Arabidopsis nitrate transporter NRT1.7,
expressed in phloem, is responsible for source-to-sink remobilization
of nitrate. Plant Cell 21, 2750–2761
26 Wang, R. et al. (1998) The Arabidopsis CHL1 protein plays a major
role in high-affinity nitrate uptake. Proc. Natl. Acad. Sci. U.S.A. 95,
15134–15139
27 Liu, K.H. et al. (1999) CHL1 is a dual-affinity nitrate transporter of
Arabidopsis involved in multiple phases of nitrate uptake. Plant Cell
11, 865–874
28 Morère-Le Paven, M-C. et al. (2011) Characterization of a dual-affinity
nitrate transporter MtNRT1.3 in the model legume Medicago
truncatula. J. Exp. Bot. 62, 5595–5605
29 Orsel, M. et al. (2002) Nitrate transport in plants: which gene and
which control? J. Exp. Bot. 53, 825–833
30 Forde, B.G. (2000) Nitrate transporters in plants: structure, function
and regulation. Biochim. Biophys. Acta. 1465, 219–235
31 Okamoto, M. et al. (2006) High-affinity nitrate transport in roots of
Arabidopsis depends on expression of the NAR2-like gene AtNRT3.1.
Plant Physiol. 140, 1036–1046
32 Orsel, M. et al. (2006) Characterization of a two-component high-
affinity nitrate uptake system in Arabidopsis. Physiology and
protein–protein interaction. Plant Physiol. 142, 1304–1317
33 Orsel, M. et al. (2007) Nitrate signaling and the two component
high affinity uptake system in Arabidopsis. Plant Signal. Behav. 2,
260–262
34 Yong, Z. et al. (2010) Characterization of an intact two-component
high-affinity nitrate transporter from Arabidopsis roots. Plant J. 63,
739–748
35 Feng, H. et al. (2011) Multiple roles of nitrate transport accessory
protein NAR2 in plants. Plant Signal. Behav. 6, 1286–1289
36 Laugier, E. et al. (2012) Regulation of high-affinity nitrate uptake in
roots of Arabidopsis depends predominantly on posttranscriptional
control of the NRT2.1/NAR2.1 transport system. Plant Physiol. 158,
1067–1078
37 Li, W. et al. (2007) Dissection of the AtNRT2.1:AtNRT2.2 inducible
high-affinity nitrate transporter gene cluster. Plant Physiol. 143,
425–433
465
Review
38 Orsel, M. et al. (2002) Analysis of the NRT2 nitrate transporter family
in Arabidopsis. Structure and gene expression. Plant Physiol. 129,
886–896
39 Chopin, F. et al. (2007) The Arabidopsis AtNRT2.7 nitrate transporter
controls nitrate content in seeds. Plant Cell 19, 1590–1602
40 Cerezo, M. et al. (2001) Major alterations of the regulation of root
NO3 uptake are associated with the mutation of Nrt2.1 and Nrt2.2
genes in Arabidopsis. Plant Physiol. 127, 262–271
41 Kiba, T. et al. (2012) The Arabidopsis nitrate transporter NRT2.4
plays a double role in roots and shoots of nitrogen-starved plants.
Plant Cell 24, 245–258
42 von der Fecht-Bartenbach, J. et al. (2010) CLC-b-mediated NO3 /H+
exchange across the tonoplast of Arabidopsis vacuoles. Plant Cell
Physiol. 51, 960–968
43 De Angeli, A. et al. (2006) The nitrate/proton antiporter AtCLCa
mediates nitrate accumulation in plant vacuoles. Nature 442,
939–942
44 Geelen, D. et al. (2000) Disruption of putative anion channel gene
AtCLC-a in Arabidopsis suggests a role in the regulation of nitrate
content. Plant J. 21, 259–267
45 Zifarelli, G. and Pusch, M. (2009) Conversion of the 2 Cl /1 H+
antiporter ClC-5 in a NO3 /H+ antiporter by a single point
mutation. EMBO J. 28, 175–182
46 Bergsdorf, E.Y. et al. (2009) Residues important for nitrate/proton
coupling in plant and mammalian CLC transporters. J. Biol. Chem.
284, 11184–11193
47 Wege, S. et al. (2010) The proline 160 in the selectivity filter of the
Arabidopsis NO3 /H+ exchanger AtCLCa is essential for nitrate
accumulation in planta. Plant J. 63, 861–869
48 Vahisalu, T. et al. (2008) SLAC1 is required for plant guard cell S-type
anion channel function in stomatal signalling. Nature 452, 487–491
49 Negi, J. et al. (2008) CO2 regulator SLAC1 and its homologues
are essential for anion homeostasis in plant cells. Nature 452,
483–486
50 Geiger, D. et al. (2011) Stomatal closure by fast abscisic acid signaling
is mediated by the guard cell anion channel SLAH3 and the receptor
RCAR1. Sci. Signal. 4, ra32
51 Lee, S.C. et al. (2009) A protein kinase–phosphatase pair interacts
with an ion channel to regulate ABA signaling in plant guard cells.
Proc. Natl. Acad. Sci. U.S.A. 106, 21419–21424
52 Wolt, J.D., ed. (1994) Soil Solution Chemistry, John Wiley & Sons
53 Jackson, R.B. and Caldwell, M.M. (1993) The scale of nutrient
heterogeneity around individual plants and its quantification with
geostatistics. Ecology 74, 612–614
54 Wang, R. and Crawford, N.M. (1996) Genetic identification of a gene
involved in constitutive, high-affinity nitrate transport in higher
plants. Proc. Natl. Acad. Sci U.S.A. 93, 9297–9301
55 Crawford, N.M. and Glass, A.D.M. (1998) Molecular and physiological
aspects of nitrate uptake in plants. Trends Plant Sci. 3, 389–395
56 Meharg, A. and Blatt, M. (1995) NO3 transport across the plasma
membrane of Arabidopsis thaliana root hairs: kinetic control by pH
and membrane voltage. J. Membr. Biol. 145, 49–66
57 Tong, Y. et al. (2005) A two-component high-affinity nitrate uptake
system in barley. Plant J. 41, 442–450
58 Huang, N.C. et al. (1996) CHL1 encodes a component of the low-
affinity nitrate uptake system in Arabidopsis and shows cell type-
specific expression in roots. Plant Cell 8, 2183–2191
59 Deane-Drummond, C.E. (1986) A comparison of regulatory effects of
chloride on nitrate uptake, and of nitrate on chloride uptake into
Pisum sativum seedlings. Physiologia Plantarum 66, 115–121
60 Doddema, H. and Telkamp, G.P. (1979) Uptake of nitrate by mutants
of Arabidopsis thaliana, disturbed in uptake or reduction of nitrate.
Physiologia Plantarum 45, 332–338
61 Touraine, B. and Glass, A.D. (1997) NO3 and ClO3 fluxes in the
chl1-5 mutant of Arabidopsis thaliana. Does the CHL1-5 gene encode
a low-affinity NO3 transporter? Plant Physiol. 114, 137–144
62 Glass, A.D.M. et al. (1992) Studies of the uptake of nitrate in barley:
IV: electrophysiology. Plant Physiol. 99, 456–463
63 Siddiqi, M.Y. et al. (1990) Studies of the uptake of nitrate in barley: I.
Kinetics of 13NO3- influx. Plant Physiol. 93, 1426–1432
64 Vidmar, J.J. et al. (2000) Isolation and characterization of HvNRT2.3
and HvNRT2.4, cDNAs encoding high-affinity nitrate transporters
from roots of barley. Plant Physiol. 122, 783–792
466
Trends in Plant Science August 2012, Vol. 17, No. 8
65 Muños, S. et al. (2004) Transcript profiling in the chl1-5 mutant of
Arabidopsis reveals a role of the nitrate transporter NRT1.1 in the
regulation of another nitrate transporter, NRT2.1. Plant Cell 16,
2433–2447
66 Lejay, L. et al. (1999) Molecular and functional regulation of two NO3
uptake systems by N- and C-status of Arabidopsis plants. Plant J. 18,
509–519
67 Fraisier, V. et al. (2000) Constitutive expression of a putative high-
affinity nitrate transporter in Nicotiana plumbaginifolia: evidence for
post-transcriptional regulation by a reduced nitrogen source. Plant J.
23, 489–496
68 Wirth, J. et al. (2007) Regulation of root nitrate uptake at the
NRT2.1 protein level in Arabidopsis thaliana. J. Biol. Chem. 282,
23541–23552
69 Liu, K.H. and Tsay, Y.F. (2003) Switching between the two action
modes of the dual-affinity nitrate transporter CHL1 by
phosphorylation. EMBO J. 22, 1005–1013
70 Ho, C.H. et al. (2009) CHL1 functions as a nitrate sensor in plants. Cell
138, 1184–1194
71 Fu, H-H. and Luan, S. (1998) AtKUP1: a dual-affinity K+ transporter
from Arabidopsis. Plant Cell 10, 63–74
72 Kim, E.J. et al. (1998) AtKUP1: an Arabidopsis gene encoding high-
affinity potassium transport activity. Plant Cell 10, 51–62
73 Newstead, S. et al. (2011) Crystal structure of a prokaryotic
homologue of the mammalian oligopeptide-proton symporters,
PepT1 and PepT2. EMBO J. 30, 417–426
74 Shi, L. et al. (2008) The mechanism of a neurotransmitter: sodium
symporter: inward release of Na+ and substrate is triggered by
substrate in a second binding site. Mol. Cell 30, 667–677
75 Daniel-Vedele, F. and Chaillou, S. (2005) Nitrogen. In Plant
Nutritional Genomics (Broadley, M.R. and White, P.J., eds), pp. 1–
25, Blackwell
76 Segonzac, C. et al. (2007) Nitrate efflux at the root plasma membrane:
identification of an Arabidopsis excretion transporter. Plant Cell 19,
3760–3777
77 Aslam, M. et al. (1995) Effect of pH and calcium on short-term NO3
fluxes in roots of barley seedlings. Plant Physiol. 108, 727–734
78 Macduff, J.H. and Jackson, S.B. (1992) Influx and efflux of nitrate
and ammonium in Italian ryegrass and white clover roots:
comparisons between effects of darkness and defoliation. J. Exp.
Bot. 43, 525–535
79 Allen, S. and Smith, J.A.C. (1986) Ammonium nutrition in Ricinus
communis: its effect on plant growth and the chemical composition of
the whole plant, xylem and phloem saps. J. Exp. Bot. 37, 1599–1610
80 Shelp, B.J. (1987) The composition of phloem exudate and xylem sap
from broccoli (Brassica oleracea var. italica) supplied with NH4+,
NO3+ or NH4NO3. J. Exp. Bot. 38, 1619–1636
81 Dodd, I.C. et al. (2003) Do increases in xylem sap pH and/or ABA
concentration mediate stomatal closure following nitrate deprivation?
J. Exp. Bot. 54, 1281–1288
82 Zhang, H. et al. (1996) Potato nitrogen management by monitoring
petiole nitrate level. J. Plant Nutr. 19, 1405–1412
83 Keisling, T.C. et al. (1995) Using cotton petiole nitrate–nitrogen
concentration for prediction of cotton nitrogen nutritional status on
a clayey soil. J. Plant Nutr. 18, 35–45
84 De Angeli, A. et al. (2009) ATP binding to the C terminus of the
Arabidopsis thaliana nitrate/proton antiporter, AtCLCa, regulates
nitrate transport into plant vacuoles. J. Biol. Chem. 284, 26526–
26532
85 Guo, F.Q. et al. (2003) The nitrate transporter AtNRT1.1 (CHL1)
functions in stomatal opening and contributes to drought
susceptibility in Arabidopsis. Plant Cell 15, 107–117
86 Castaings, L. et al. (2009) The nodule inception-like protein 7
modulates nitrate sensing and metabolism in Arabidopsis. Plant J.
57, 426–435
87 Alboresi, A. et al. (2005) Nitrate, a signal relieving seed dormancy in
Arabidopsis. Plant Cell Environ. 28, 500–512
88 Wang, R. et al. (2009) A genetic screen for nitrate regulatory mutants
captures the nitrate transporter gene NRT1.1. Plant Physiol. 151,
472–478
89 Walch-Liu, P. and Forde, B.G. (2008) Nitrate signalling mediated by
the NRT1.1 nitrate transporter antagonises l-glutamate-induced
changes in root architecture. Plant J. 54, 820–828
Review Trends in Plant Science August 2012, Vol. 17, No. 8

90 Remans, T. et al. (2006) The Arabidopsis NRT1.1 transporter
participates in the signaling pathway triggering root colonization
of nitrate-rich patches. Proc. Natl. Acad. Sci. U.S.A. 103, 19206–19211
91 Krouk, G. et al. (2010) Nitrate-regulated auxin transport by NRT1.1
defines a mechanism for nutrient sensing in plants. Dev. Cell 18, 927–937
92 Tian, Q.Y. et al. (2009) Ethylene is involved in nitrate-dependent root
growth and branching in Arabidopsis thaliana. New Phytol. 184,
918–931
93 Hachiya, T. et al. (2010) Evidence for a nitrate-independent function
of the nitrate sensor NRT1.1 in Arabidopsis thaliana. J. Plant Res.
124, 425–430
94 Hachiya, T. and Noguchi, K. (2011) Mutation of NRT1.1 enhances
ammonium/low pH-tolerance in Arabiopsis thaliana. Plant Signal.
Behav. 6, 706–708
95 Meraviglia, G. et al. (1996) The chl1 Arabidopsis mutant impaired in
the nitrate-inducible NO3 transporter has an acidic intracellular pH
in the absence of nitrate. J. Plant Physiol. 149, 307–310
96 Guo, F.Q. et al. (2001) The Arabidopsis dual-affinity nitrate
transporter gene AtNRT1.1 (CHL1) is activated and functions in
nascent organ development during vegetative and reproductive
growth. Plant Cell 13, 1761–1777
97 Remans, T. et al. (2006) A central role for the nitrate transporter
NRT2.1 in the integrated morphological and physiological responses
of the root system to nitrogen limitation in Arabidopsis. Plant Physiol.
140, 909–921
98 Little, D.Y. et al. (2005) The putative high-affinity nitrate transporter
NRT2.1 represses lateral root initiation in response to nutritional
cues. Proc. Natl. Acad. Sci. U.S.A. 102, 13693–13698
99 Camañes, G. et al. (2012) A deletion in NRT2.1 attenuates
Pseudomonas syringae-induced hormonal perturbation, resulting in
primed plant defenses. Plant Physiol. 158, 1054–1066
100 Zhao, X. et al. (2010) Genomic survey, characterization and expression
profile analysis of the peptide transporter family in rice (Oryza sativa
L.). BMC Plant Biol. 10, 92–111
101 Lin, C.M. et al. (2000) Cloning and functional characterization of a
constitutively expressed nitrate transporter gene, OsNRT1, from rice.
Plant Physiol. 122, 379–388
102 Li, S. et al. (2009) Short panicle1 encodes a putative PTR family
transporter and determines rice panicle size. Plant J. 58, 592–605
103 Loque, D. et al. (2003) Gene expression of the NO3 transporter NRT1.1
and the nitrate reductase NIA1 is repressed in Arabidopsis roots by
NO2 , the product of NO3 reduction. Plant Physiol. 132, 958–967
104 Guo, F.Q. et al. (2002) The Arabidopsis dual-affinity nitrate
transporter gene AtNRT1.1 (CHL1) is regulated by auxin in both
shoots and roots. J. Exp. Bot. 53, 835–844
105 Lejay, L. et al. (2003) Regulation of root ion transporters by
photosynthesis: functional importance and relation with
hexokinase. Plant Cell 15, 2218–2232
106 Filleur, S. and Daniel-Vedele, F. (1999) Expression analysis of a high-
affinity nitrate transporter isolated from Arabidopsis thaliana by
differential display. Planta 207, 461–469
107 Wang, R. et al. (2007) Nitrite acts as a transcriptome signal at
micromolar concentrations in Arabidopsis roots. Plant Physiol. 145,
1735–1745
108 Lejay, L. et al. (2008) Oxidative pentose phosphate pathway-
dependent sugar sensing as a mechanism for regulation of root ion
transporters by photosynthesis. Plant Physiol. 146, 2036–2053
109 Okamoto, M. et al. (2003) Regulation of NRT1 and NRT2 gene families
of Arabidopsis thaliana: responses to nitrate provision. Plant Cell
Physiol. 44, 304–317
110 Zhuo, D. et al. (1999) Regulation of a putative high-affinity nitrate
transporter (Nrt2;1At) in roots of Arabidopsis thaliana. Plant J. 17,
563–568
111 Nazoa, P. et al. (2003) Regulation of the nitrate transporter gene
AtNRT2.1 in Arabidopsis thaliana: responses to nitrate, amino acids
and developmental stage. Plant Mol. Biol. 52, 689–703
112 Filleur, S. et al. (2001) An Arabidopsis T-DNA mutant affected in Nrt2
genes is impaired in nitrate uptake. FEBS Lett. 489, 220–224

Plant Science Conferences in 2012

11th Nordic Photosynthesis Congress NPC11
11 – 14 September, 2012
Naantali, Finland
http://npc11.utu.fi/

30th New Phytologist Symposium

Immunomodulation by plant-associated organisms
16 – 19 September 2012
California, USA
http://www.newphytologist.org/immunomodulation/default.htm

4th International EcoSummit 2012

30 September – 5 October
Columbus, USA
http://www.ecosummit2012.org/

VI International Conference on Legume Genetics and Genomics

2 – 7 October, 2012
Hyderabad, India
http://www.icrisat.org/gt-bt/VI-ICLGG/Homepage.htm

467