Auxins
RM Napier, University of Warwick, Coventry, UK
2017 Elsevier Ltd. All rights reserved.
This article is a revision of the previous edition article by R.M. Napier, volume 2, pp. 985995, 2003, Elsevier Ltd.
Introduction
Auxins are natural plant hormones, mobile biochemicals,
which are transported from cell to cell and organ to organ to
signal and coordinate growth and development. The experi-
ments of Charles Darwin on phototropic bending by grass cole-
optiles in the 1880s led him to conclude that plants contained
a mobile inuence mediating growth. In the 1920s, Fritz Went
collected diffusates from coleoptiles and showed that the
inuence induced bending when applied to unstimulated
coleoptiles. Shortly afterward, in 1934, Kgl identied
indole-3-acetic acid (IAA) as the active compound.
The main natural auxin in all plants is IAA, and it is synthe-
sized primarily in dividing meristematic cells. Auxin is an abso-
lute requirement for plant cell division, but through directed
and nondirected transport, it also mediates control over
many other key events during development. These include
asymmetric growth during phototropism and gravitropism,
apical dominance, fruit set and development, abscission,
both lateral and adventitious root initiation, phyllotaxy, and
vascular patterning. It is not surprising, therefore, that auxin
concentrations in the plant are carefully controlled, that there
is a sophisticated machinery for polar auxin transport, and
that some microorganisms have capitalized on their own
ability to synthesize auxins in order to perturb a host plants
development and exploit the disturbance for either pathogen-
esis or symbiosis. It is also clear that addition of auxin to plants
will affect growth, and synthetic auxins are used widely in
a range of agricultural and horticulture practices. The largest
single market for auxins is as selective herbicides, but auxins
are also applied as agrochemicals on fruit crops and for root
induction during clonal propagation of nursery stocks.
Auxins
In addition to IAA, a small number of naturally occurring,
related compounds have auxin activity. These include
4-chloroindole-3-acetic acid, phenylacetic acid and, possibly,
indole-3-butyric acid (IBA). Recent advanced metabolic
proling of auxinic compounds found no trace of IBA in several
plant species, although earlier biological and mass spectrom-
etry data supported its presence. If it is produced in planta,
IBA is probably converted to IAA by b-oxidation in plant cell
peroxisomes, making the active form of this compound IAA.
A much more heterologous group of synthetic compounds
are found to have auxin activity. Some of the most widely used
are listed in Table 1.
IAA Measurement, Synthesis, and Breakdown
Measurement
Detailed measurements of auxin synthesis and of total auxin
concentrations are becoming routine. As the technology
develops, real value is being added by the simultaneous
Encyclopedia of Applied Plant Sciences, 2nd edition, Volume 1 http://dx.doi.org/10.1016/B978-0-12-394807-6.00099-X
measurement of the auxin metabolome. This includes quantica-
tion not only of IAA, but also of its precursors, metabolites, and
catabolites. Extracts are passed through a solid phase extraction/
purication column before the eluate is concentrated by drying
under vacuum. After being dissolved in a small volume of meth-
anol, compound separation and quantication uses liquid
chromatography-multiple-reaction-monitoring-mass spectrom-
etry. Unfortunately, these exacting analytical techniques are avail-
able in very few laboratories. Protocols are also developing for
parallel analyses of a number of plant hormones to give data
about antagonistic and complementary signals from the same
small samples. Only the auxin metabolome is dealt with here.
Auxin Conjugates and Storage
For many years it was thought that IAA was synthesized in apical
meristems. However, recent evidence suggests that auxin is, or
can be, produced throughout the plant, including the roots.
Furthermore, not all IAA production is from de novo synthesis.
In germinating seeds, IAA is produced from breakdown of stored
forms of the hormone, conjugates of amino acids, proteins, and
sugars. Broadly, monocotyledonous plants (cereals and grasses)
accumulate sugar conjugates, dicotyledonous plants accumulate
amino acid conjugates. Hydrolysis of these conjugates during
germination precedes or coincides with the start of root exten-
sion. For example, large uxes of IAA released from conjugates
stored in the kernel mediate extension in cereal coleoptiles.
This auxin moves in the vascular strands to the coleoptile tip
and is then distributed back down to the elongating cells by
the polar auxin transport system (see below).
De Novo Synthesis
Stored IAA reserves are nite and soon after seedling emergence
de novo IAA synthesis commences. The amino acid tryptophan
is the precursor for a set of parallel IAA biosynthetic pathways
(Figure 1). Probably the most important pathway outside the
Arabidopsis (Brassica) family is the conversion of tryptophan
to indole-3-pyruvic acid by a family of tryptophan amino-
transferases (TAAs). The YUCCA family of avin monooxyge-
nase enzymes (YUCs) then converts indole-3-pyruvate to IAA.
Indole-3-pyruvate may also be decarboxylated to give indole-
3-acetaldehyde and then converted to IAA by indoleacetalde-
hyde hydroxylase. In Brassica species, YUCCA enzymes are
also involved in the oxidation of tryptamine to indole-3-
acetaldoxime. Cytochrome P450s convert this to indole-3-
acetonitrile as well as other secondary compounds, but these
pathways appear absent from other plant families.
A few days after germination, a further biosynthetic
pathway is activated, the tryptophan-independent pathway.
The intermediates on this pathway have not been identied,
but tryptophan is not one of them. It seems likely that this
tryptophan-independent pathway remains the constitutive
biosynthetic source of IAA throughout the rest of a plants
life. However, when a plant is stressed, such as by wounding
or during major developmental events like germination, IAA
367
368 Regulators of Growth j Auxins

Table 1 Common auxins

Abbreviation or
Auxin Formula product name Uses

Indole-3-acetic acid H IAA Natural auxin

N

OH

Indole-3-butyric acid HN IBA Natural auxin. Rooting compound

O

OH

2,4-Dichlorophenoxyacetic acid Cl OH 2,4-D Plant tissue culture. Selective herbicide

Cl O O

3-Amino-2,5-dichlorobenzoic acid H2N Cl Chloramben Selective herbicide, preemergence

Amiben
OH

O
Cl

Picloram Cl Tordon Selective herbicide

N OH
Cl
O
H2N Cl

Triclopyr Cl OH Garlon Selective herbicide, orchard fruit

anti-abscission agent
Cl O O
N
Cl

Quinclorac Cl Quinclorac 75 Selective herbicide, postemergence

N Cl

O OH

Auxin transport inhibitors

1-Naphthylphthalamic acid O NPA Auxin efux inhibitor

HN
O
HO

2,3,5-Triiodobenzoic acid I I TIBA Transport inhibitor and weak auxin

OH

O
I
 Regulators of Growth j Auxins 369

Figure 1 Auxin distribution in a mature Arabidopsis plant. The distribution of free IAA was measured using gas chromatography/mass spectrometry.
Concentrations are illustrated by color-coding the organs; the scale is shown to the right. Reproduced with permission from Mller, A., Dchting, P.,
Weiler, E.W., 2002. A multiplex GC-MS/MS technique for the sensitive and quantitive single-run analysis of acidic phytohormones and related
compounds, and its application to Arabidopsis thaliana. Planta 216, 4456.

synthesis is stepped up by switching on the tryptophan-
dependent pathways.
Very young leaves, less than 0.5 mm long in Arabidopsis,
produce large amounts of IAA (Figure 1). These early high
concentrations and high synthetic capacity help drive leaf
expansion, but as the leaf expands, synthesis and concentra-
tions fall. Concentrations of free IAA were found to be
100-fold lower in expanded leaves. In fully grown Arabidopsis
plants the highest concentrations of IAA are found in expand-
ing fruiting bodies, the siliques. It is likely that much of this
is synthesized by seeds during embryo development (see
below). The inorescence stem contains more IAA than most
other parts of the plant, but it is not clear if this is synthesized
in situ or if it is in transit.
Deactivation and Homeostasis
Control of auxin action can be mediated by removing IAA as
well as by synthesis. Auxins are removed by oxidation and by
conjugation into inactive storage compounds. For example,
IAA concentrations elevated experimentally are rapidly moder-
ated by oxidation of IAA to 2-oxindole-3-acetic acid and conju-
gation to both amino acid (mainly aspartic acid) and glucose
conjugates. Some enzymes involved in these processes are
known, including very recently a dioxygenase for auxin oxida-
tion, but their regulation genetic or biochemical is poorly
understood.
The balance of synthesis, breakdown, conjugation, and
transport is regulated rigorously to give auxin homeostasis.
Leaf expansion, for example, is reduced by both increases
and decreases of auxin concentration. Clearly, changes in
auxin concentration are important as plants respond to
stimuli, but homeostasis is critical for both optimal develop-
ment and to keep the system primed for stimulatory
responses. The best-characterized enzyme system which
contributes to homeostasis is the GH3 family of IAA-amido
synthetases, which use ATP to catalyze formation of amino
370 Regulators of Growth j Auxins
Figure 2 Pathways of auxin synthesis, metabolism, and catabolism. The main intermediate compounds are given in frames, the principal enzymes
in shaded boxes.
acid conjugates (Figure 2). Glutathione S-transferases play
a similarly important role for synthetic auxins, reacting the
applied compounds to glutathione, and these inactive
conjugates are rapidly moved out of the cytoplasm into the
vacuole. Other enzymatic feedback mechanisms for
homeostasis are not well characterized, but intracellular
compartmentation driven by both IAA and IAA-conjugate
transport proteins will contribute (see below).
Inhibitors of Auxin Synthesis and Metabolism
A growing interest in novel chemical tools for manipulating
auxin physiology has led to screens for stage-specic inhibi-
tors. Yucasin, an analogue of indole-3-pyruvic acid, is
a competitive inhibitor of YUCCA and Lkynurenine, an
analogue of tryptophan is a competitive inhibitor of TAAs.
The structure of one GH3 has been solved by freezing its
mechanism using an inhibitory reaction intermediate
analogue in the active site. Such novel compounds have not
found commercial applications yet, but are proving useful
new experimental tools.
Nonplant Auxin Synthesis
The auxin IAA is made by certain microorganisms as well as by
plants, but it appears that only plants can control auxin
concentrations to give auxin homeostasis. Additionally, it
appears that only plants have auxin receptors and respond
to auxin as a hormone, although there is some evidence that
plant nematodes can sense auxin and use this as a signal to
move toward a new host. Bacteria such as the pathogens Pseu-
domonas syringae and Agrobacterium tumefaciens synthesize
copious amounts of auxin from tryptophan, making use of
a pathway not exploited by plants. A tryptophan monooxyge-
nase (iaaM) converts tryptophan to indole-3-acetamide,
which is converted by a hydrolase (iaaH) to IAA. Such auxin
contributes to the pathogenic responses induced in plant
hosts, such as in the crown gall tumors caused by agrobacte-
rial infections. Likewise, there is evidence to suggest that
symbiotic mycorrhizal fungi produce auxin, and that this
auxin then contributes to the changing morphology of the
root system in these associations.
For many years it was thought that plants did not use the
iaaM/H pathway, possibly because it allowed insufcient
control over the production of this important hormone.
Recently, plant homologues have been identied (Figure 2).
Auxin Transport
All plant hormones are moved around the plant in the vascula-
ture where, once loaded, they move passively as passenger
molecules. Auxin is special in that plants also have an active
(energy requiring) polar transport system to move IAA direc-
tionally, as well as additional auxin transport mechanisms for
both intracellular and intercellular movements. Polar auxin
transport contributes to many of the vital auxin-dependent
morphogenic responses described below.
Auxin Uptake and AUX1
Polar transport works at the cellular level and is affected by the
combined activities of both auxin inux proteins and auxin
efux proteins (Figure 3). The inux carrier is known as
 Regulators of Growth j Auxins 371

Figure 3 Auxin transport at the cellular level. Most IAA molecules are protonated at the pH of the cell wall (pH 5.56). Protonated IAA is membrane
permeant and so may diffuse freely into cells. The cytoplasm is pH 77.4 and so IAA converts rapidly into the IAA-anion. This is not membrane
permeant and so is trapped inside the cell. There are also uptake carrier proteins in the plasma membrane to assist in accumulating free IAA. Inside
the cell there are carrier proteins to move IAA and IAA-conjugates into subcellular compartments such as the endoplasmic reticulum. There are also
carrier proteins to pump IAA out of the cell, and both PINs and ABCBs contribute to directional auxin ow, polar auxin transport.

AUX1, and it was identied from genetic screens using seed-
lings insensitive to the auxin 2,4-D. The AUX1 protein is
a member of the amino acid transporter superfamily. Trans-
port is driven by the carriage of two protons into the cell
with each IAA molecule. AUX1 is an integral membrane
protein spanning the lipid bilayer with 10 or 11 helices joined
by short loops of hydrophilic amino acids. Members of this
protein family are well characterized in bacteria, with both
structure and transport mechanism solved, and so it may
not be long before we have similar detail about this important
plant transporter.
Auxin Efux and PIN Proteins
The rst efux carrier gene was identied from plants display-
ing an extreme ower morphology. Pin-like organs are formed
with no branching or further development of the inores-
cence. This morphology gave rise to the family name PIN,
and there are more than 10 PIN genes in Arabidopsis. Indi-
vidual members of the PIN gene family are expressed in
dened sets of cells and tissues throughout the plant. The
proteins themselves sit in the plasma membrane, span the
membrane 10 times, and act to pump auxin out of cells.
Critically, they are localized asymmetrically around the cell
such that efux activity induces polar auxin ow (Figure 3).
By doing so, PINs not only transmit the auxin signal but
contribute to the information content by effecting, for
example, auxin gradients.
A family of PIN-like proteins known as PILS has been
shown to have auxin transport activity, and these are
intracellular, sited on the endoplasmic reticulum and other
endomembranes where they contribute to auxin homeostasis.
Other Auxin Transporters
The speed of polar auxin transport is generally measured at
520 mm h1. Under circumstances in which auxin might be
moved through few cells or cell layers to effect a gradient for
differential growth (such as for gravitropism in roots or coleop-
tiles), and for small plants, such a transport rate seems sustain-
able. However, the passage of an auxin stimulus from the apex
of a tree to its roots presents a challenge of a different magnitude.
Observations have been made of a wave of auxin release trav-
eling in advance of, and more rapidly than, net polar auxin trans-
port. The mechanisms for such a wave have still to be identied,
but merit further research.
372 Regulators of Growth j Auxins
It is not correct to surmise that all auxin movement is
through the polar transport system. Considerable quantities
of free IAA and of auxin conjugates are carried in the vascula-
ture, particularly the phloem. Indeed, the relatively high pH
in phloem acts to concentrate free IAA, and unloading from
the phloem will require the action of carrier proteins like PIN
and AUX1, although the mechanisms regulating unloading
sites remain to be identied.
Additional auxin transport proteins have also been
described, and the most important of these is the ATP-
binding cassette pump proteins known as ABCBs, although
some older literature also refers to them as PGPs. Related to
pump proteins involved in, for example, multidrug tolerances
in animals, there is a family of 21 in Arabidopsis. These show
tissue- and cell-specic expression, and specic members are
present on different membranes in the cell, contributing to
both long- and short-distance polar auxin ow and to
intracellular auxin compartmentalization.
Two additional membrane transporters have been added to
the auxin carrier list. WAT1 is a vacuolar auxin transporter,
moving auxin out of the vacuole. When decient, the secondary
plant cell walls of xylem wood cells are uncharacteristically
thin. Auxin uptake from the apoplast is facilitated by a trans-
porter known for its activity as a nitrate carrier and nitrate
sensor. NRT1 has been shown to carry auxin as well as nitrate,
although not together and nitrate inhibits auxin transport
through NRT1.
Inhibitors of Auxin Transport
Many of the auxin-driven responses considered below fail if
auxin is absent or in excess, or if polar auxin transport is defec-
tive. It is clear that hormone delivery is as important as the
hormone itself. As a result, the study of auxin physiology has
beneted from the use of drugs that act specically to inhibit
polar auxin transport. Of these, naphthylphthalamic acid
(NPA) (Table 1) is the most specic and has been used widely.
It has also been shown that NPA binds to certain ACBC carrier
proteins, although this does not exclude other sites of action on
other transporters. Triiodobenzoic acid (TIBA) has also been
used, but has additional activities. A useful tool to inhibit
uptake has been identied as 1-naphthoxyacetic acid, and
researchers have started to compare and contrast the
structure-activity proles of transporters alongside, for
example, receptors in order to separate the detailed mecha-
nisms contributing to auxin phenotypes.
Auxin in Plant Development
Vascular Patterning
Auxin transport inhibitors have dramatic effects on the devel-
opment of the vasculature in developing tissues. Auxin deter-
mines both sites of vascular differentiation and promotes
vascular connectivity. Most work has been done in leaves and
using Arabidopsis, in which auxin transport inhibitors lead to
connement of the vasculature to the leaf margins. The zone
of cell division in leaves lies at the margins, and auxin synthesis
in very young leaves is extremely active, as noted above. It
seems likely that auxin from the margins induces vascular
Figure 4 (a) Typical leaf venation in a young Arabidopsis leaf. (b) Heat
map of auxin gradients. Red shows high auxin concentrations, mainly
around the peripheral meristematic cells. Blue shows low auxin in the
vascular streams. As soon as a new vascular strands forms local auxin
concentrations drop. Prior to vascular connection local auxin maxima
form and the gradient from these maxima to the vascular sink deter-
mines the development of new vasculature, as illustrated in the panel
marked*.
development. The midvein and petiole hold existing vascula-
ture and act as a sink leading to an auxin gradient from source
at the edge to sink at the center of the leaf (Figure 4). This
gradient lays down the axis of early patterning. As new vascula-
ture forms, it generates additional auxin sinks and the new
phloem exports IAA, reinforcing local gradients. Both before
and during vascular development, polar transport provides
directionality and there are convincing computer models
backed up by impressive microscopy showing the relationship
between auxin gradients, developing polarization of PIN trans-
port proteins, and the consequent reinforcement of both polar
auxin ow and tissue patterning (see below).
Patterns of primary and secondary vascular strands in devel-
oping leaves are set very early, and once set, they are not
changed by auxin stimuli. In addition to synthesis at the leaf
margins it has been suggested that specialized leaf cells in the
lamina, such as glandular hydathodes, provide auxin and
help determine tertiary patterning. It can be seen that
combining these various axes and gradients of auxin, along
with other stimuli, gives a complex set of developmental cues
which result in diverse, yet reproducible, venation patterns.
Embryonic Patterning
Just as polar ow of auxin is essential for cell axis formation in
the vasculature, it plays the same, key role in establishment of
the plant body axis in young embryos. For example, auxin trans-
port inhibitors prevent the establishment of bilateral symmetry
in embryos. Auxin responses during embryogenesis have been
well studied using antibodies to describe the organization of
PIN proteins from the earliest embryos (Figure 5) combined
with genetic reporters for auxin action, which are driven by the
auxin-sensitive promoter element known as DR5. Auxin direc-
tional ow has been deduced from the polar distribution of
PINs, and cells high in auxin are documented with color or uo-
rescence from DR5 reporters. These approaches have been
augmented with detailed models of early developmental stages
of a plants life. Morphogenic Arabidopsis mutants have also

Figure 5 Patterning in the very young embryo is determined by the
polarity of PIN auxin transport proteins. (a) In the 2-cell embryo, auxin
is pumped by PIN7 proteins up into the embryonic stem cells to help
drive cell division. (b) In the slightly later embryo, at the early to midg-
lobular phase, the apical meristem niche is already synthesizing IAA
and now acts as a source of auxin (purple). The polarity of the cell at
the neck of the embryo (the hypophysis) reverses and PIN1 and PIN4
proteins now pump IAA away from the developing vegetative embryo,
aided by PIN7 at the base of this hypophysis cell at the neck. The
hypophysis will start to differentiate into root tissue.
been useful tools. One known as MONOPTEROS shows the
same embryonic failure to establish a polar body axis as seen
after NPA treatment. The MONOPTEROS gene codes for an
auxin-regulated transcription factor, a protein controlling gene
expression. Mechanisms of auxin action and the control of
gene expression are considered below.
Organ Patterning
It is clear that auxin and its polar transport determine the
apicalbasal pattern of development throughout the plant at
every scale. For example, in oral tissues such as the gynoe-
cium, the specication of the few stylar cells through which
the pollen tube will pass on its way to fertilize the ovule is
auxin-driven, and the ancestral event which led to the evolu-
tion of owering plants (angiosperms) may well have been
control over the placement of PIN proteins to secure radial
auxin gradients in the gynoecium. (The role of auxin in fruit
development is discussed below.)
Perhaps the most widely observed outcome of auxin-
induced patterning is in phyllotaxis, the regular arrangement
of leaves on a stem. Once again, polar auxin transport inhibi-
tors and Arabidopsis mutants defective in polar transport like
pin1-1 have helped illustrate that accumulation of IAA in cells
at the side of the shoot apical meristem initiates organogenesis
and determines the position of the next leaf primordium. Some
delicate and beautiful experiments have been described by the
group of Kuhlemeier in Switzerland. Microapplications of
transport inhibitors or IAA to tomato (Lycopersicon esculentum)
shoot apices have shown that, for leaves and owers, auxin foci
determine primordium initiation in the radial dimension. In
the natural situation, auxin transport proteins in the surface
cell layers route auxin toward the initiation site of the next
primordium, and these local foci ensure that the next
Regulators of Growth j Auxins 373
Figure 6 Auxin determines phyllotaxy. A lateral view of a tomato
shoot apical meristem showing two leaf primordia, P3 to the right at
the rear, P2 to the left at the rear, and P1, the area at the face of the
apical dome (z). Auxin delivery that is dependent on polar auxin trans-
port is indicated by the red arrow. This auxin causes local expression of
organ identity and outgrowth genes. Lateral cellcell signaling leads to
boundaries (y) and inhibition of organogenesis (x). Reproduced with
permission of Elsevier Ltd., from Kuhlemeier, C., Reinhardt, D., 2001.
Auxin and phyllotaxis. Trends Plant Sci. 6, 187189.
development always arises at a xed distance from the summit
of the apical meristem in the apicalbasal dimension
(Figure 6).
Tropisms and Epinasty
Plants respond to the vectoral signals light and gravity by
differential growth. Auxin gradients generated by lateral (polar)
auxin transport drive bending in both shoots and roots. The
experiments of Cholodny and Went using grass or cereal seed-
lings are common schoolroom lessons. Blocks of agar were
used to collect endogenous signals from two sides of gravisti-
mulated seedling, from just below the tip. The blocks were
then applied to unstimulated seedlings and shown to induce
appropriate bending. Application of exogenous auxins
achieved the same result.
In stems, greater auxin concentrations enhance elongation
growth (in roots, high auxin concentrations inhibit elongation
growth). Auxin measurements have shown that the side of stems
expanding more rapidly contains a little more IAA, although the
gradient across the stem is generally only of the order of two- or
threefold. Auxin reporter genes show expression patterns corre-
sponding to small auxin gradients (Figure 7), but even so it is
argued that the magnitude of the gradient is insufcient to
support the considerable differences in elongation rate across
the same tissues. Recent data on the stimulus-induced redistribu-
tion of specic members of the PIN family, data from auxin
374 Regulators of Growth j Auxins
Figure 7 Auxin distribution after gravistimulation in roots. Expression
of an auxin-responsive reporter gene (DR5-GUS) is observed by the
accumulation of blueness. The vertical root tip is rich in auxin, but the
cells behind the tip show little or no expression, indicating low auxin
concentrations. However, if the root tip is turned through 90 to lie hori-
zontal, auxin is seen to accumulate in cells along the lower side in the
root elongation zone. In roots, high concentrations of auxin inhibit cell
elongation, and so the cells on the upper side elongate to push the root
tip back toward the vertical. Reformatted from the original; reproduced
with permission from Rashotte, A.M,, DeLong, A., Muday, G.K., 2001.
Genetic and chemical reductions in protein phosphatase activity alter
auxin transport, gravity response, and lateral root growth. Plant Cell 13,
16831697. Copyrighted by the American Society of Plant Biologists.
measurements, and observations from auxin response mutants
all support the CholodnyWent hypothesis of auxin-driven
extension growth, but auxin ux may be as important as the pre-
vailing IAA concentration. Other data, such as kinetic measure-
ments of gravistimulated roots and experiments in which
seedling shoots are bathed in excess IAA yet are still found
able to respond by differential growth, argue against such
a straightforward hypothesis. Either way, it is clear that auxin
is a requirement for both differential growth and tropic growth,
although it might not be the only signal involved.
A similar phenomenon to tropic growth is epinasty, petiole
bending induced by a range of nondirectional stimuli. The top
sides of the petioles extend excessively, leading to curvature
with the result that the face of the leaf often turns away from
light and leaves look twisted. The contribution auxin plays in
epinasty is seen most clearly in plants treated with auxinic
herbicides (see below), but the phenomenon is also associated
with plant stress, particularly root ooding. Other hormones,
in particular ethylene, are also involved in epinasty.
Apical Dominance and Branching
In addition to control over the initiation of leaf primordia at
shoot apices, it has been recognized for many years that auxin
transported down from the shoot apex inhibits the outgrowth
of side branches from axillary buds. Gardeners and horticultur-
alists make use of this apical dominance to control plant struc-
ture by pruning. The classical story is based on the loss of the
primary source of auxin with removal of the stem apex. Before
excision, this auxin passed down the plant by polar auxin trans-
port, inhibiting lateral outgrowth from axillary buds. Removal
of this inhibitory signal leads to side branch outgrowth and
bushier plants. Consistent with these utilitarian observations,
classical experiments by Thimann and Skoog showed that if
auxin is applied to the pruned apical stump, lateral bud
outgrowth is inhibited. More recently, experimental plants
generated with low-endogenous auxin levels are bushier,
consistent with the idea that high auxin inhibits axillary bud
growth. However, apically derived auxin does not enter the
axillary bud, and it is unclear how the signal is transferred.
Root-derived cytokinins are also involved in controlling bud
break and other rootshoot signals such as the strigolactones
play key roles, but a counter hypothesis to the classic polar
auxin ow story presents sugar demand as the primary regu-
lator. As well as the apex providing a constant and strong
supply of auxin, it requires a great deal of energy to grow.
Hence, decapitation cancels this sugar sink and releases the
sugar for use by the next intact meristem.
Clearly auxin contributes to the complex of signaling events
during branch initiation, but it may not be the principal player
in axillary bud outgrowth. A further character associated with
plant architecture is gravitropic set-point angle. Every plant
species has not only a characteristic (genetically encoded) phyl-
lotaxy to determine the angle between leaves as they arise at the
shoot apical meristem (and hence branches which arise as axil-
lary buds), but also a characteristic set-point angle with respect
to gravity the angle between main shoot and each main
branch, for example. As noted above, auxin redistribution
drives the extension growth needed to restore the axis of growth
during reorientation with respect to gravity. In order to arrest
this reorientation or to set the normal growth angle, there
needs to be a compensating signal at the opposite side of the
organ. Surprisingly, this is also auxin, although how auxin
gradients are set, and how receptor systems are gauged to
respond remaining areas of active research.
Root Development
Apical dominance is active in roots as well as stems, but for
roots the contribution of additional specic signals is less clear.
The root tip certainly acts as an important site for lateral redis-
tribution of auxin. Some is generated in situ, as at the shoot
apical meristem, but a major supply is also delivered down
the stele. Auxin transporters, both uptake and efux carriers,
are critical for managing the reverse ow of IAA back up the
plant from the root apex in the outer cell layers, notably the
epidermal layer. This reverse ow determines tropic responses,
elongation growth, etc., and in roots, high auxin concentrations
inhibit growth. However, increases in auxin do drive initiation
and outgrowth of lateral roots.
Lateral root development is controlled by auxin at several
stages. In the root tip just behind the elongation zone lateral
primordia form, but remain dormant. These primordia will
not grow out unless the tip is removed (apical dominance)
or until a second, shoot-derived, polar ux of auxin arrives.
Not long after emergence a lateral root becomes auxin-
autonomous.
Cell Division, Tissue Culture, and Secondary Meristems
So far, only the inuence of auxin on tissue development has
been considered. However, one of the most profound actions
of auxin on plants is over the control of cell division. In the
absence of auxin, or at low concentrations, plant cells will
not divide. The cell cycle arrests at G1. Cell expansion can
proceed at low auxin concentrations, but only at elevated

concentrations will the cell cycle advance past G1 to G2/M and
division. During division, cell expansion is minimal.
Among the most striking examples of auxin control over
division is in cell and tissue culture for which auxin must be
included in almost all media. Furthermore, auxin (and cyto-
kinin) concentrations are manipulated to promote proliferative
callus growth, vegetative regeneration, and root induction. For
root induction, auxin concentrations are raised; for regenera-
tion, auxin concentrations are reduced, cytokinin concentra-
tions are raised.
Secondary meristems are important for the generation of
many plant tissues. Among the most prolic is the cambium
which gives rise to secondary xylem and phloem, and the
tissues of the phellem (bark). Auxin concentrations have
been measured in thin, sequential sections across the cambial
region of pine trees, from inner xylem (wood) to the bark. The
highest concentration corresponded with cambial cells, and it
has been suggested that this zone of high auxin conveys posi-
tional information. In this way auxin is acting as
a morphogen, and the steepness of the declining gradient of
auxin across cells adjacent to the cambium determines the
radial width of differentiating zones of division and expan-
sion. As for cell division (high IAA) and cell expansion (low
IAA), there are concentration thresholds which combine
with other positional stimuli to mediate the length of time
cambial derivatives spend dividing, differentiating, or laying
down secondary walls. Consequently, much secondary
growth is governed by auxin concentrations in, and delivery
from, the cambium.
It has been reported that all cambial auxin is delivered by
polar auxin transport, mostly from the expanding stem apices.
Polar transport inhibitors deplete almost all the auxin from the
cambial region. However, considering the speed of polar auxin
transport (see above), it is recognized that there are additional
sources of IAA, and the dividing cambium might itself
contribute to the pool.
Adventitious Rooting and Wound Responses
Adventitious roots form from stem tissues, generally as a result
of damage or removal of the primary root system. Many plants
do produce some adventitious or crown roots naturally, and
they add valuable lateral support to tall stems in cereals, for
example. However, addition of auxin to cut or damaged stems
often induces a strong adventitious rooting response. The horti-
cultural industries relying on clonal propagation make good use
of this response and annual sales of clonal ornamentals, trees,
owers, and other garden plants continue to rise around the
world. Synthetic auxins IBA and 1-naphthylacetic acid (1-NAA)
are both sold widely in hormone rooting powders and dips.
For lateral primordia to form in primary root tissues, cells of
the root xylem pericycle divide and differentiate. For adventi-
tious rooting, a similar series of events is likely to take place
in equivalent cells in the stem vasculature, but the exact cells
giving rise to the rst division are less certain. New root
primordia form at the side of the procambium or within callus
tissue that forms at the cut or wounded surface. The initiation
of root primordia in stem tissues requires a redifferentiation
response and, although auxin promotes this, the molecular
or genetic mechanisms are unclear.
Regulators of Growth j Auxins 375
There are mutants of Arabidopsis that show precocious root-
ing along the seedling hypocotyl, superroot 1 and 2 (sur1 and
sur2), for example. In each, auxin levels are elevated due to
mutations in genes associated with secondary metabolite
production from tryptophan and other intermediates. By pre-
venting secondary metabolite synthesis, intermediates build
up and feed an acceleration of auxin biosynthetic pathways.
Wounding plant tissues give rise to rapid cell division and
callus formation. Callus is undifferentiated and apolar in its
growth, but not only can new, polar organs arise from it (like
adventitious roots), but adjacent cells can redifferentiate to
reform vascular connectivity. The process is slow, commencing
many days after damage and callus growth starts, but xylem
elements soon start to form with a polar orientation, lining
up to reestablish new vasculature as fully formed, thickened
xylem elements die. Auxin and polar auxin transport are essen-
tial for this response, and it is likely that this polar redifferentia-
tion occurs along a reforming apicalbasal auxin gradient. This
is not a general event, this redifferentiation occurs only in
a narrow band of cells that starts and nishes at damaged
vascular strands. A similar process is likely to take place during
graft development so that vascular continuity is regained.
The reformation of vascular connectivity has been studied
as an archetypal and auxin-mediated response to reestablish
tissue polarity. Tsvi Sachs developed a highly inuential cana-
lization theory of auxin ow, and this continues to enervate
models of auxin-mediated differentiation, all driven by auxin
gradients establishing cell polarities and auxin ow via directed
accumulations of PIN proteins (e.g., Figures 4 and 5).
Fruit Growth
Auxin plays a vital role in all stages of reproductive growth. The
role of auxin in gynoecium development and embryo polarity
has been recorded above, and some of the highest auxin
concentrations have been found to be in developing fruit
(Figure 1). In the early 1950s, Nitsch showed that achene
(seed) removal from strawberry (Fragaria  ananassa) recepta-
cles inhibited receptacle enlargement. Replacing the achene
with a supply of auxin maintained fruit growth. Cessation of
auxin supply also led to ripening in this nonclimacteric fruit.
In climacteric fruit, ripening is mediated by ethylene not
auxin. Nevertheless, in a wide range of fruit as well as in cereal
seed-heads, it is clear that the developing embryo and seed are
rich auxin sources, and that this auxin mediates many develop-
mental responses in adjacent tissues. Indeed, parthenocarpy
can be induced in many species by auxin application, and
this has been used to generate seed-free fruit by both topical
application and by genetic engineering. The bacterial IAA
biosynthetic gene iaaM coding for tryptophan monooxygenase
was transformed into tomato, eggplant (Solanum melongena,
aubergine), and other fruit under the control of a carpel-
specic promoter. This gave rise to localized overproduction
of IAA and induced seed-free fruit of marketable quality. The
need for bees to pollinate in the glasshouse was removed,
a problem during winter months, and so there was an added
advantage that production was no longer seasonal. Some apple
(Malus pumila) and cherry (Prunus spp.) crops are sprayed with
auxin at owering to induce fruit set, although there is gener-
ally a mix of hormones used.
376 Regulators of Growth j Auxins
Fruit crops, particularly Citrus spp., have been sprayed with
auxins to improve fruit size by promoting June drop but, more
widely, orchards were sprayed after the drop to increase fruit
size and hexose content, and to delay fruit abscission. Addition
of auxin delays the development of the abscission zone. Both
exocarp and seeds are rich in IAA, but, as in strawberries, it is
likely that as the testa matures around the seed it forms an
impervious barrier, blocking auxin release from these internal
stores. In the absence of auxin, fruit abscission proceeds. Leaf
abscission is also initiated as auxin transport from the leaf
declines.
Treatment of orchards to delay fruit abscission became
common to facilitate automated harvesting, maximizing the
yield of ripe fruit collected in the minimum number of passes
by harvesters. Lychee (Litchi chinensis) and nut crops are also
treated with auxins to promote fruit size and stall abscission.
However, such use on minor crops (horticultural crops as
opposed to dominant arable crops) is seldom promoted by
producers due to the high costs of licensing compounds for
minor markets. Nevertheless, harvest uniformity and quality
are key traits for plant breeders, and improved variety choices
are likely to take the place of some agronomic auxin
applications.
Commercial Importance of Auxins
Herbicides
The largest commercial exploitation of any plant hormone has
been in the use of synthetic auxins as selective herbicides.
Around the world a number of populations of auxinic herbi-
cidetolerant weeds have arisen, an indication of the long
time over which applications have been made. The rst phe-
noxyacetic acid auxins, such as 2,4-D (Table 1), were developed
in the 1940s and use has been extensive ever since. Farmers
have exploited their selectivity, for they have a much higher
activity against most dicotyledonous plants than against
cereals, and this had led to widespread use on cereal crops,
sugarcane, and turf to control broadleaved weeds. The auxin
2,4,5-T was the active defoliant in Agent Orange, but not the
toxin (a dioxin). Phenoxy auxins remain in widespread use,
but auxins with much lower dose requirements are being adop-
ted increasingly for agricultural use.
The mechanisms of herbicidal action and the basis of selec-
tivity between monocotyledons and dicotyledons are unclear.
Induction of a massive synthesis of ethylene has been suggested
to be the mechanism of activity, at least in part because a by-
product of ethylene overproduction is cyanide. However, recent
work shows that inhibition of either ethylene synthesis or
ethylene receptors does not prevent herbicidal activity. There-
fore, although induction of the ethylene-synthesizing enzymes
is a recognized effect of auxin application, ethylene remains
a symptom of herbicidal auxin and is not the sole mediator.
Auxinic herbicides also induce abscisic acid (ABA)
synthesis, and this might, or might not, be a consequence of
elevated ethylene synthesis. Ethylene and ABA act antagonisti-
cally in growth control. Elevated ABA will shut stomata, and the
consequent block on photosynthesis might be the source of
excessive reactive oxygen production, as recorded in auxinic
herbicide-treated plants. In addition to the damage done by,
for example, peroxide itself, such reactive oxygen species will
induce defense and senescence responses, giving rise to necrosis
and death. As noted above, a familiar symptom of auxinic
herbicide treatment is strong epinasty, and it is likely that
uncontrolled surplus of a synthetic auxin overloads many of
the delicate gradients discussed above leading to miscommuni-
cation and death.
Saturating auxin response systems clearly gives rise to
a number of damaging symptoms, but genetic data from resistant
biotypes, as well as the observations of monocotyledon/dicoty-
ledon selectivity suggest that there is a single target site for these
herbicides, and this seems likely to be the auxin receptor.
Auxin Perception, Receptors, and Signaling
Perception and Receptors
Many techniques have been used in experiments to identify
auxin receptors, and many candidates have been examined in
detail. Photoactive and radiolabeled auxins have been shown
to bind to a diverse set of proteins, afnity purication has
identied others, and genetic screens have yielded a vast
amount of information on auxin transport proteins, signaling
intermediates, and on the transcriptional regulation of auxin-
induced genes. Yet there is still scope for the discovery of novel
receptors.
The rst candidate auxin receptor was identied in the
early 1970s in membrane fractions of Zea coleoptiles. It was
characterized as a protein, which bound active auxins and is
referred to as auxin-binding protein 1 (ABP1). Purication
and sequencing of ABP1 followed in the late 1980s. It is
a soluble protein targeted to the endoplasmic reticulum.
The three-dimensional crystal structure of ABP1 with and
without auxin bound has been determined at high resolution,
giving the rst images of a plant hormone recognition site.
Over many years, experiments to test the function of ABP1
linked it to functions at the plasma membrane, such as
membrane polarization, ion currents, and endocytosis.
However, new ABP1 genetic knockout lines have recently
cast doubt over all the phenotypes associated with ABP1
and its activity as a receptor for auxin. These lines do not differ
measurably from wild-type plants. Earlier lines described as
ABP1 knock-outs, and reported to be embryo-lethal, have
been shown to contain lesions that affect both ABP1 expres-
sion and an adjacent, essential gene. The new lines suggest
convincingly that ABP1 is not a receptor, and its role remains
unresolved.
Conrmed receptors for auxin are the TIR1 and the AFB
proteins. These, and associated proteins, were discovered
from genetic screens seeking mutants insensitive to auxins
and auxin transport inhibitors. The rst to be cloned and
sequenced was a mutant known as axr1. The sequence data sug-
gested the gene product coded for an ortholog of a ubiquitin-
activating enzyme and, therefore, the AXR1 protein was likely
to be involved in protein degradation. Subsequently, many
other components of ubiquitin ligase complexes and the pro-
teasome were revealed from additional auxin-insensitive
mutant lines, and it was clear that the key developmental
events described above involved auxin-mediated transcrip-
tional control via degradation of transcriptional regulators.

Figure 8 (a) Crystal structure of TIR1 (pale blue) showing ASK1
(dark blue) at the base and bound to the F-box domain of TIR1. The
leucine-rich repeats made up of serial alpha helices, and beta sheets
form a ring with the ends locking against each other. In the center of
the ring is a single molecule of inositol-6-phosphate (mauve) which
also helps to hold the ring intact. The part of the leucine-rich repeat
structure directly involved in binding IAA (red) is colored purple, and
when the IAA pocket is occupied the coreceptor and ubiquitination
target AUX/IAA binds over the top (orange). Only a peptide
representing the degron domain of an Aux/IAA protein is shown.
(b) The residues lining the auxin-binding pocket (purple in (a)) in
detail and with the synthetic auxin 2,4-D shown bound. Polar residues
(green) form the base of the pocket, hydrophobic residues (blue)
form a ring around the aromatic core of the bound auxin. The degron
(not shown) will cover and ll the remaining space in the pocket,
trapping auxin at the base.
TIR1 and its family AFBs are nuclear proteins. Their structure
has been solved by crystallography, again with and without auxin
bound. There is a large ring of leucine-rich repeats, and an adja-
cent domain known as an F-box domain which associates these
F-box proteins to the rest of a ubiquitin E3 ligase complex via
the scaffold proteins ASK1 and CUL1. The auxin binding site is
at the base of a pocket lined by a series of the leucine-rich
repeats (Figure 8). When IAA binds, it creates a new surface
within the binding pocket, and this new face is a binding site
for the degron domain of a family of transcriptional regulators
known as the Aux/IAA proteins. By binding, these Aux/IAA
proteins become ubiquitinated and destined for degradation by
the proteasome. These reactions are rapid and the half-life of
many Aux/IAAs is between 10 and 15 min.
In the absence of auxin, Aux/IAAs are mostly found dimer-
ized with another family of transcriptional regulators, the auxin
response factor (ARF) proteins. Originally discovered due to
their selective binding to the promoter regions of auxin-
regulated genes, ARFs alone (probably as homodimers) are
transcriptional activators. Heterodimerized with Aux/IAAs,
they are repressed. Hence, with the appearance of auxin, Aux/
IAAs are ubiquitinated and degraded, and transcriptional
repression is released. Some of the earliest products of this tran-
scriptional burst are new Aux/IAAs, giving sensitive feedback
control over the system.
The defective gene in the embryo morphology mutant
MONOPTEROS, mentioned above, is a member of the Aux/
IAA gene family. A mutant with some similar morphological
features, BODENLOS, is an ARF. Despite the undoubted
contribution to plant form and function played by auxin
transport proteins, there is no doubt that the interpretation
of auxin stimuli and auxin gradients is played out by receptors
and the immediate targets of their activation. A large part of
auxin action is governed by the control of gene activation
Regulators of Growth j Auxins 377
driven by Aux/IAA ubiquitination and breakdown. Given
the number of ARFs and Aux/IAAs and the large number of
different, mixed dimers that can be made, it is seen that this
system can account for much of the great variety of different
responses controlled by auxin. However, linking each
response to a dened pair of transcriptional regulators will
take considerable effort.
Summary
There are still gaps in our knowledge of auxin synthesis,
signaling, and action, but a great deal has been learned. Many
of the wonderful plant forms and functions are mediated by
precise and ephemeral movements of auxin. These gradients
of IAA are interpreted by the TIR1 family of receptor proteins,
and the detailed structural information available is now allow-
ing exploration of still further auxinic molecules. There are
many agriculturally useful applications of both auxins and of
our growing knowledge of how auxin phenotypes are deter-
mined. As we learn more about auxin genetics and biochem-
istry, a still wider set of applications can be anticipated.
Acknowledgments
Funding has been provided by BBSRC award BB/L009366.
See also: Arable Crops: Field Crops. Horticulture Production
and Quality: Orchard Crops. Plant Breeding and Genetics:
Molecular Biology of Development; Transformation and
Transgene Expression. Plant Cells: Cell Division and Cell
Differentiation; Cell Growth; Leaf Development; Wood Growth
and Development. Plant Nutrition: Genetics of Primary Root
Development; Growth and Function of Root Systems; Lateral
Root Initiation. Postharvest Biology: Flower Senescence;
Genetic Engineering for Postharvest Quality; Leaf Senescence;
Ripening. Regulators of Growth: Abscisic Acid; Cytokinins;
Gibberellins. Reproduction and Biodiversity: Fertilization. Seed
Development and Germination: Embryogenesis. Tissue
Culture: General Principles of Tissue Culture; Organogenesis;
Somatic Embryogenesis. Weeds and Competition: Gene Flow
and Herbicide Resistance.
Further Reading
Berleth, T., Mattsson, J., Hardtke, C.S., 2000. Vascular continuity and auxin signals.
Trends Plant Sci. 5, 387393.
Davies, P.J., 1995. Plant Hormones: Physiology, Biochemistry and Molecular Biology.
Kluwer Academic, New York.
Korasick, D.A., Enders, T.A., Strader, L.C., 2013. Auxin biosynthesis and storage
forms. J. Exp. Bot. 64, 25412555.
Leyser, H.M.O., 2010. The power of auxin in plants. Plant Physiol. 154, 501505.
Mason, M.G., Ross, J.J., Babst, B.A., Weinclaw, B.N., Beveridge, C.A., 2014. Sugar
demand, not auxin, is the initial regulator of apical dominance. Proc. Natl. Acad.
Sci. U.S.A. 111 (16), 60926097.
Yoshida, S., Saiga, S., Weijers, D., 2013. Auxin regulation of embryonic root formation.
Plant Cell Physiol. 54 (3), 325332.
Zazimalova, E., Petrasek, J., Benkova, E., 2014. Auxin and Its Role in Plant Devel-
opment. Springer, Wein.