Auxin: The First Discovered Plant

Growth Hormone
 Contents
 What is hormone ?
 Types of hormone
 Plant’s hormone
 Introduction to Auxin
 Discovery of Auxin
 The Principle Auxin: Indole-3-Acetic acid
 IAA Synthesis
 Different pathways
 Contents
 Auxin Transport
 Chemiosmotic model of molar Auxin transport
 Auxin influx
 Auxin efflux
 Auxin signal transduction pathway
 Action of Auxin
 Cell elongation
 Plant tropisms
 Developmental effects of Auxin
 References
 WHAT IS HORMONE ?
• Hormones are chemical messengers that are
produced in one cell and modulate cellular
processes in another cell by interacting with
specific proteins that function as receptors
linked to cellular transduction pathway.
 Types of hormones
Endocrine hormones
• Hormones that are
transported to sites of
action in tissues distant
from their site of synthesis
are referred to as endocrine
hormones
Paracrine hormones
• Hormones that act on cells
adjacent to the source of
synthesis are referred to as
paracrine hormones.
 Plant hormones
• The plant development is regulated by six
major hormones:
• Auxins
• Gibberellins
• Cytokinins
• Ethylene
• Abscisic acid
• Brassinosteroids
 Plant hormones
• There are some signaling molecules
(hormones) like jasmonic acid, salicylic acid and
small polypeptides that play roles in resistance
to pathogens and defense against herbivores.
• Strigolactone regulates the outgrowth of lateral
buds and stimulate the branching and growth
of symbiotic  arbuscular mycorrhizal 
fungi, (helps in capturing nutrients and minerals.
 Introduction to Auxin
• First growth hormone to be studied in plants
• Play important role in growth and
development of plant
• Developmental processes like stem
elongation, apical dominance, root initiation,
fruit development, meristem development is
controlled by auxin
 Introduction to Auxin

• Auxin and cytokinin are required for viability

of the plant embryo and organogenesis
• Whereas other hormones act as regulators of
discrete developmental processes
 The Emergence of the Auxin concept
• Charles Darwin and his son Francis studied plant
growth phenomena involving tropisms
• Their interest of study was the bending of plant
towards light
• The phenomenon, which is caused by differential
growth, is called phototropism
• Studying on Seedlings of canary grass ( Phalaris
canariensis)
• Young leaves (newly emerging leaves) are in protective
sheath called coleoptile
 The Emergence of the Auxin concept

• Coleoptiles are highly sensitive to light

• The Darwins hypothesized that shoots bend
towards light in response to an “influence”
transmitted downward from the tip

• After that , Peter Boysen-Jensen and Arpad Paal

demonstrated that the “influence” was actually
a chemical

11
 The Emergence of the Auxin concept
In 1926, Frits Went performed an experiment and
studied growth promoting chemicals in tips of Avena
sativa that explained all of the previous results
-He named the chemical messenger auxin
-It accumulated on the side of an oat seedling
away from light
-Promoted these cells to grow faster than
those on the lighted side
-Cell elongation causes the plant to bend
towards light
13
15
 The Principle Auxin: Indole-3-Acetic acid

• In the mid 1930,it was determined that natural

auxin is indole-3-acetic acid
• Various other auxins are found in plant
• Auxins are the compound with biological activities
similar to those IAA
• Cell elongation in coleptile and stem section
• Cell division in callus cultures
• Formation of adventitious roots on detached
leaves and stem
 Types of Auxins
Natural Auxins :
indole-3-acetic acid (IAA)
4-chloroindole-3-acetic acid (4-Cl-IAA), phenylacetic acid (PAA)
indole-3-butyric acid (IBA)
 Syntyetic auxin

2,4-Dichlorophenoxyacetic acid
α-Naphthalene acetic acid

2,4,5-Trichlorophenoxyacetic acid

2-Methoxy-3,6-dichlorobenzoic acid 4-Amino-3,5,6-trichloropicolinic acid

 IAA synthesis
• IAA biosynthesis is associated with rapidly
dividing and growing tissues
• Primary site of auxin synthesis are shoot apical
meristem and young leaves
• Root apical meristem also synthesize auxin
• Young fruits and seeds also have high level of
auxin
 IAA synthesis
• In Arabidopsis, auxin accumulate at tip of
young leaves primordia
• In developed leaves move towards the margin
• Then shift at base of leaf
• Tryptophan is the probable precursor of auxin
biosynthesis.
 Biosynthesis Pathways

• Multiple pathways
• Mostly two types of pathways exist
 Tryptophan –dependent pathway
 Tryptophan –independent pathway
 Tryptophan dependent pathway
• IAA structurally related to Amino Acid Tryptophan

• 3 different tryp.dependent
 The IPA pathway
 The TAM pathway
 The IAN pathway
 Trytophan
IPA IAN
Deamination TAM

Decarboxylation

Indole-3-acetaldoxime
Indole-3-pyruvic acid Tryptamine
Oxidation and
Deamination
Decarboxylation Indole-3-acetonitrile
Indole -3-acetaldehyde

Oxidation

IAA
 IPA (Indol -3- pyruvic acid) Pathway

• It is probably the most common of the tryptophan-

dependent pathways.
• It involves a deamination reaction to form IPA,
followed by a decarboxylation reaction to form
indole-3-acetaldehyde (IAld).
• Indole-3-acetaldehyde is then oxidized to IAA by a
specific dehydrogenase.
 TAM (Tryptamine) Pathway

• It is similar to the IPA pathway, except that the

order of the deamination and decarboxylation
reactions is reversed, and different enzymes are
involved.
• Species that do not utilize the IPA pathway
possess the TAM pathway.
• Although in one case (tomato), there is evidence
for both the IPA and the TAM pathways.
•
 TAM (Tryptamine) Pathway

STEPS
• Tryptophan is converted to TAM by tryptophan
decarboxylase .
• TAM then converted to Indole acetaldehyde by
amine oxidase (oxidation of amines)
• Finally IAA is formed from IALD by
dehydrogenase enzyme
 The IAN(indole-3-acetonitrile)
Pathway

• The IAN pathway may be important in only three

plant families: the Brassicaceae (mustard family),
Poaceae (grass family), and Musaceae (banana
family).
• Nevetheless, nitrilase like genes or activities have
recently been identified in Cucurbitaceae (squash
family), Solanaceae (tobacco family), Fabaceae
(pea family) and rosaceae (rose family).
The IAN(indole-3-acetonitrile) Pathway

STEPS:
• Tryptophan is first converted to indole-3-
acetaldoxime and then to indole-3-
acetonitrile.
• The enzymes responsible for this conversion
are still unknown.
• The enzyme that converts IAN to IAA is called
nitrilase
 The IAM (indole-3-acetamide) Pathway

• Another tryptophan-dependent biosynthetic

pathway—one that uses indole-3-acetamide (IAM)
as an intermediate is used by various pathogenic
bacteria, such as Pseudomonas savastanoi and
Agrobacterium tumefaciens.
• This pathway involves the two enzymes tryptophan
monooxygenase and IAM hydrolase.
• The auxins produced by these bacteria often elicit
morphological changes in their plant hosts.
Biosynthesis of Auxin
 Tryptophan independent pathway

• Recent genetic studies have provided evidence

that plants can synthesize IAA via one or more
tryp.independent pathways.
• E.g., synthesis of IAA from indole or Indole -3-
glycerol phosphate.
• Evidence came from orange pericarp mutant of
maize (orange color of pericarp
• of maize kernel is due to the
• excessive indole)
 Tryptophan independent
pathway
• orp mutant is a true tryptophan auxotroph
(unable/ defective in tryptophan synthesis b/c of
lack of tryptophan synthase enzyme).
• However, neither orp seedlings nor wild type
seedlings can convert trp to IAA.
• Despite block in trp biosynthesis, orp mutant
contains IAA 50-fold higher than those of wild
type.
•
 Evidence of trp-independent pathway
Isotopic [15N]anthranilate

Applied exogeneously to orp mutant

Labelled susbsequently appeared in IAA

Not in tryptophan

Lead to the evidence of trp independent pathway

 Tryptophan independent pathway

• Further studies establish that the branch point for

IAA biosynthesis is either indole or indole 3-
glycerol phosphate
• IAN & IPA are possible intermediates
• But the immediate precursor of IAA in trp-ind
pathway not yet identified.
 Tryptophan
independent
pathway
 Auxin transport
• Transport polarly
• Main axes of shoots and roots have apex-base
structural polarity
• Went discovered that IAA moves mainly from
the apical to basal end (basipetal).
• This type of unidirectional transport is termed
polar transport
A Chemiosmotic Model Has Been Proposed to
Explain Polar Transport
Polar transport require energy and is gravity
independent
Polar transport require energy and is gravity
independent
• Polar transport proceeds in a cell to cell fashion
• The auxin exits the cell through the plasma
membrane, diffuses across the compound middle
lamella and enters the next cell through its plasma
membrane
• Export of auxin from cell is termed auxin efflux
• Entry of auxin into cell is called auxin uptake or
influx
• The overall process requires metabolic energy
 Polar transport require energy and is gravity
independent

• The velocity of polar auxin transport is 5-20cmh-¹

• faster than the rate of diffusion
• slower than phloem translocation rates
• Polar transport is also specific for active auxins, both natural
and synthetic.
• Neither inactive auxin analogs nor auxin metabolites are
transported polarly,
• polar transport involves specific protein carriers on the plasma
membrane that can recognize the hormone and its active
analogs.
A Chemiosmotic Model Has Been Proposed to
Explain Polar Transport
• The discovery in the late 1960s
• According to the chemiosmotic model for
polar auxin transport,
• auxin uptake is driven by the proton motive
force (PMF) across the plasma membrane,
• while auxin efflux is driven by the membrane
bounded efflux proteins.
A Chemiosmotic Model Has Been Proposed to
Explain Polar Transport
 Inhibitors of Auxin Transport Block Auxin
Efflux

• Several compounds have been synthesized that can

act as auxin transport inhibitors (ATIs), including
• NPA (1-Nnaphthylphthalamic acid)
• TIBA (2,3,5-triiodobenzoic acid)
• CPD( 2- carboxyphenyl-3-phenylpropane-1,3-dione)
• NOA ( naphthoxyacetic acid)
 Inhibitors of Auxin Transport Block Auxin
Efflux

• NPA,TBA, and CPD are auxin efflux inhibitors

• NOA is an auxin influx inhibitor
• AEIs can interfere with the trafficking of the
plasma membrane proteins to which they
bind, apparently by altering protein
interactions.
 Non polar Transport
• Recently, it has been recognized that a
significant amount of auxin transport also
occurs in the phloem
• Phloem is the principal route for acropetal
(i.e., towards the tip) transport.
• Most of the IAA that is synthesized in mature
leaves, appears to be transported to the rest
of the plant nonpolarly via phloem
• Passive, not require energy.
 Auxin Signal Transduction
 Most likely, multiple auxin perception sites are
present such as :
• ABP 1 (auxin binding protein), ABP57
• TIRI 1 (Transport inhibitor response 1)
 Different families of proteins are involved in
auxin-induced changes in gene expression
– Auxin response factors (ARFs)
• Have DNA binding domains
• Can enhance or suppress transcription of genes
 Auxin Signal Transduction
– Aux/IAA Repressor proteins
• Bind and repress ARF proteins that activate the
expression of auxin related genes
– TIR1
• is the auxin receptor
– Ubiquitin protein
• Small protein
• Binds to a protein (as a tag) for degradation via
26S proteasome
• Prevents protein –protein interactions
 Auxin and gene regulation
• In the absence of auxin:
• Aux / IAA (repressor protein) bind to the ARF
• Prevents its binding to promoter site
• No gene expression
• In the presence of auxin:
• Aux binds to SCF TIR1
• tag AUX/ IAA repressor protein (attach with ARF)
with ubiquitin ligase and then degraded
• Active dimer of ARF formed and gene is expressed
50
AUXIN SIGNAL TRANSDUCTION
PATHWAYS
 Auxin induced genes
• Auxin induced gene fall in to two classes :early and late
• One of the important functions of the signal transduction
pathways initiated when auxin bind to its receptor is the
activation of a selective group of transcription factors
• The activated transcription factors enter the nucleus and
promote the expression of specific genes
• Genes whose expression is stimulated by the
activation of pre existing transcription factors are
called primary response genes or early genes
 Functions of Early genes
1) regulate the transcription of secondary response
genes
2) involved in intercellular communication , or cell to
cell signaling
3) another group of early genes is involved in
adaptation to stress
• Early genes expression cannot be blocked by
inhibitors of protein synthesis such as cyclo heximide
• Expression of early genes is very short time ranging
from few minutes to several hours
Fig 11. A MODEL FOR EARLY AUXIN REGULATED GENE EXPRESSION AND GENE
FUNCTION (Ranjan et al.,2003)

Auxin

Other
inducer
Transcriptional Activation of early genes

Aux RE
ACS SAUR Aux/IAA GH3 GH2/4

?
Glutathione S -
ACC
Transferase
synthase Short lived
nuclear protein Detoxicification Cellular Redox
Ethylene State Auxin Binding / Transport

Intercellular Regulation of Stress

Signaling gene expression Response
 Late /secondary genes

• Some of the early genes encode proteins that

regulate the transcription of secondary
response genes
• These are required for the long term response
to the hormones
• Late genes require de novo protein synthesis,
their expression is blocked by protein
synthesis inhibitors
 Physiological Roles

1. CELL ELONGATION

• Auxin was discovered as the hormone involved

in the bending of coleoptiles toward light
• Auxins Promote Growth in Stems and
Coleoptiles, While Inhibiting Growth in Roots
• Auxin transport basipetally from shoot apex to
the tissue below
• Auxin causes continued elongation of these cells
 CELL ELONGATION

• The optimal auxin concentration for elongation growth is

typically 10-6 to 10-5 M.
• Inhibition in growth beyond the optimal conc. Is due to
the synthesis of ethylene (inhibits stem and root
elongation).
• However, roots require minimum conc. of auxin for
elongation as compared to stem.
• Low concentrations (10-10 to 10-9 M) of auxin promote the
growth of intact roots,
• Whereas, higher concentrations (10-6 M optimal for stem
elongation) inhibit growth in roots.
 CELL ELONGATION: Auxin rapidly increases the extensibility of the cell wall

• Auxin cause a five to ten fold increase in the growth rate in only 10
minutes.
• How ?
• To understand, first review the process of enlargement in plant cells.

Plant cell expands in three steps:

1) Osmotic uptake of water across the plasma membrane is
driven by the gradient in water potential.
2) Turgor pressure builds up because of rigidity of the cell wall.
3) Biochemical wall loosening occurs, allowing the cell to expand in
response to turgor pressure.
CELL ELONGATION: Acid growth hypothesis

• 3rd step, wall loosening is related to the acidification of

cell wall
• Auxin induces proton extrusion, acidifies the cell wall,
and increases cell extension.
• According to acid growth hypothesis
– hydrogen ions act as a intermediate between the auxin and
cell wall loosening
– Cells actively transport hydrogen ions from the cytoplasm
into the cell wall space via ATPase.
– Drop in pH activates enzymes (expansins) that loosens the
cell walls by weakening the H-bonds b/w polysaccharide
components of wall
Loosening of cell wall
 CELL ELONGATION: Mechanisms of
auxin induced proton extrusion

• Auxin could increase the rate of proton

extrusion by two possible mechanisms
1) Activation of pre-existing plasma membrane
H+- ATPases
2) Synthesis of new H+- ATPases on plasma
membrane
 2. Cell differentiation
• Auxin promotes differentiation of vascular
tissue (i.e., xylem & phloem):
• Auxin and sugar -----> Vascular tissue
• Auxin and low sugar (1.5 - 2.5%) -----> Xylem
• Auxin and high sugar (4%) ------->- Phloem
• Auxin and moderate levels of sugar (2.5 -
3.0%) ----->- Xylem & Phloem
 Developmental effects of Auxin
Apical Dominance
• The growing apical bud inhibits the growth of
lateral (axillary) buds __a phenomenon called
apical dominance.
• In 1920, Skog and Thimann performed an
experiment on bean and result showed that
outgrowth of axillary bud is inhibited by auxin that
is transported basipetally
• The auxin content of bud increases by decapitation
of shoot apex
Auxin Regulates Apical Dominance
 Delays the onset of abscission
• Shedding of leaves, flowers and fruits from the
living plant --- abscission
• These parts abscise in a region called----abscission
zone
• Located near the base of petiole of leaf
• Within the abscission zone distinct layer of cells
called----abscission layer
• During senescence, walls of cells in layer digested
---become soft and weak ---leaf eventually breaks
off at abscission layer
Abscission Zone and layer
 Delays the onset of abscission
• Auxin levels high in young leaves, decreases in
maturing leaves, and are relatively low in
senescing leaves
• Removal of leaf blade ----accelerates the
formation of abscission layer in petiole.
• Application of IAA in lanolin paste (waxy
material) to the cut surface of petiole prevents
formation of abscission layer.
 Delays the onset of abscission
• These results suggest that:
1) Auxin transported from the blade normally
prevents abscission
2) Aabscission is triggered or promoted during
leaf senescence, when auxin is no longer
produced
 Auxin transport regulates floral buds

• Two experiments
1) Treating Arabidopsis with auxin transport
inhibitors NPA (efflux inhibitor)----causes abnormal
floral development.
• Suggesting that polar transport is required for
normal floral development
2) In Arabidopsis, pin 1 mutant, lacks an auxin efflux
carrier in shoot tissues----has abnormal flowers
like NPA treated plants ..
 Auxin transport regulates floral buds
• In both cases the meristem is starved /
deficient for auxin, so normal floral
development and phyllotaxis are disturbed..
• Apparently, the developing floral meristem
depends on auxin basipetal transport.
 Auxins promotes fruits development

• Evidence suggests that auxin is involved in the

regulation of fruit development
• Auxin is produced in pollen, endosperm and
embryo of developing seeds
• After fertilization, fruit growth depends on
auxin produced in developing seeds
• Experiment on strawberry explains auxin role
Auxins promotes fruits development
 Auxin Promotes the Formation of Lateral
and Adventitious Roots
• primary root is inhibited by auxin concentrations
greater than 10–8 M
• however, high concentration initiate lateral and
adventitious roots
• Lateral roots arise from small groups of cells in
the pericycle
• Auxin (transported acropetally) stimulates these
pericycle cells to divide
 Auxin Promotes the Formation of Lateral
and Adventitious Roots
• In the formation of lateral roots, auxin is required
for at least two steps
1) IAA transported acropetally in the stele is
required to initiate cell division in the pericycle
2) IAA is required to promote cell division and
maintain cell viability in the developing lateral
roots.
Adventitious roots can arise from mature cells that
renew their cell division activity.
Synthetic auxins have a variety of commercial
uses
• Prevention of fruit & leaf drop, induction of
parthenocarpic fruit development, thinnning
of fruit & rooting of cuttings for plant
propagation
• If excised leaf is dipped in auxins then rooting
enhanced
• Parthenocarpic fruit can be induced by
treatment of unpollinated flowers with auxins
Synthetic auxins have a variety of commercial
uses
• 2,4-D and dicamba are synthetic auxins which
are used as herbicides
• monocots are able to inactivate synthetic
auxins rapidly than dicots, so farmers use
synthetic auxins for the control of dicot
weeds.
 Synthetic Auxins
Naphthalene acetic acid (NAA) and
indolebutyric acid (IBA) have many uses in
agriculture and horticulture
-Prevent abscission in apples and berries
-Promote flowering & fruiting in pineapples
Alhamdulillah
Alhamdulillah