Plant Hormones

Lectures 1 and 2
 Readings
• An Introduction to Plant Physiology – Hopkins
and Hüner Wiley

• Biochemistry and Molecular Biology of Plants

– Buchanan et al ASPP

• Molecular Biology of the Cell – Alberts et al

Garland Science
• Plants are multicellular organisms and as such require
coordination between cells.

• In order to coordinate their activities cells must be able

to communicate with each other.

• The principle means of communication within plants are

the hormones.

• Hormones are signal molecules that individually or

cooperatively direct the development of individual cells
or carry information between cells and thus coordinate
growth and development.
• Although molecules that influence growth and
differentiation in plant cells have been studied
for nearly 100 years the concept of hormones
in plants is still steeped in controversy.

• The beginning of plant hormone research can

be traced to a series of experiments carried out
by Charles Darwin examining phototropism.
Charles Darwin
• Charles Darwin was among the 
first scientists to dabble in plant
hormone research.

• In his book "The Power  of

Movement in Plants" presented
in 1880, he first describes the
outcomes of  light on motion of
canary grass (Phalaris
canariensis) coleoptiles.
 It was Darwin’s and others observations and
experiments that led to F.W. Went, almost 50 years
later, to describe a hormone like substance as the
causative agent when plants grew towards the light.
 According to Current Usage
• Hormones are naturally occurring organic substances that, at
low concentrations, exert a profound influence on physiological
processes.

• In addition hormones as defined by animal physiologists are

 
• synthesized in a discrete organ or tissue

• transported in the bloodstream to a specific target tissue

where they

• control a physiological response in a concentration dependent

manner
• Plant hormones like animal hormone are
naturally occurring organic substances that
profoundly influence physiological processes
at low concentrations.

• However there are differences between plant

and animal hormones
 A
• The site of synthesis of plant hormones is not
are clearly localized as in animal systems,
synthesis of plant hormones appears to be
much more diffuse and cannot always be
localized to discrete tissues.

• Action at a distance is not an essential

property of plant hormones.
 B
• Whether plant hormones act in a concentration
dependent manner is still a subject of some
debate.

• Some scientists argue that plant cells respond

to hormone concentration; whereas others
argue that it is changing sensitivity of the target
cells to the hormone. Evidence exists for both
arguments.
 C
• Plant hormones have a multiplicity of effects.

• Each group of plant hormones is known to

influence a wide variety of developmental
events.

• Moreover each of these events can be

influenced by more than one hormone group.
Traditionally there are five recognized groups of plant hormones

• These are auxins, gibberellins, cytokinins, abscisic

acid and ethylene. More recently a sixth group,
the brassinosteroids, has been found to have well-
defined functions in normal plant development.

• Jasmonic acid and salicylic acid are elicitor

molecules that appear to be active in regulating
some aspects of plant growth and may qualify for
hormone status.
 Auxins
• Auxin was the first plant hormone to be discovered. It
is generally ubiquitous in the plant, although the
highest concentrations of the hormone are detected
in meristematic regions and actively growing regions
such as the coleoptile apices, root tips, the apical buds
and germinating seeds.

• Other regions of the plant that have high auxin levels

include young leaves, developing inflorescences and
embryos following pollination and fertilization.
• Auxins are characterized principally by the
capacity to stimulate cell elongation, but they
also influence a host of other developmental
responses including root initiation, vascular
differentiation, tropic responses and the
development of axillary buds, flowers and
fruits.
 
 Although a large number of
compounds have been discovered
with auxin activity indole-3-acetic
acid (IAA) is considered
to be the principle natural auxin.

• In addition to IAA, several other naturally occurring indole

derivatives are known to show auxin activity. These include
indole-3-ethanol, indole-3-acetaldehyde and indole-3-
acetonitrile.
• However these compounds all serve as precursors to IAA
and their activity is probably due to conversion to IAA
within the plant tissue.
• IAA the most widely distributed,
• IBA, 4 chloroindole-3-acetic acid and phenylacetic acid are
naturally occurring but less prevalent.
• 2,4,D and NAA are synthetic auxins
• Because IBA, 4-chloroIAA and PAA have all been isolated
from plants, are structurally similar to IAA and elicit
many of the same responses as IAA there is a strong
argument for considering them as natural hormones.

• However it is not yet established whether they are

converted to IAA in the plant tissue before they become
active.

• Chemically the single unifying character of molecules

that show auxin activity appears to be an acidic side
chain on an aromatic ring.
• The levels of IAA present depend on a number of
factors including the type and age of tissue and its
state of growth.

• In vegetative tissue the amount of IAA generally falls

in the range of between 1 mg and 100 mg kg-1
fresh weight (5.7 – 570 nanomoles).

• In seeds the levels of IAA appear to be much higher

with levels reaching up to 300 picomoles. This high
level of IAA may be to support the rapid growth of
the seedling following germination.
Tryptophan independent IAA Biosynthesis (Biological
Importance Remains Unclear)
 Auxin Deactivation
• Oxidative decarboxylation of IAA is known to be catalyzed by
peroxidase from a variety of plant sources.

• There are differing requirements for enzymes from different

sources and even multiple isozymes within a single species.

• However decarboxylative oxidation of IAA has been studied

most extensively with purified HRP in vitro. Because the end
products of IAA oxidation are physiologically inactive, IAA
oxidation is an effective way of removing the hormone
molecule once it has accomplished it purpose.
• The enzyme may either catalyze the
oxidation of the indole ring, forming an
oxindole, or decarboxylate the acetate
side chain without oxidizing the indole
ring.

• Which of the two pathways proceeds

depends on a number of factors.
However the major end product of the
oxindole route 3-methylene oxindole has
not been detected in plants in vivo, while
indole-3-carboylic acid, indole-3-
aldehyde and indole -3- methanol have
all been isolated from plants.

• This would suggest that the

decarboxlated indole pathway is the
most likely normal catabolic pathway in
plants.
 Auxin Conjugates
• For many years, decarboxylative catabolism was
thought to be the major IAA degradation pathway.
• However evidence obtained with Z. mays, pea and
tomato suggest that peroxidases have only a minor
role in the regulation of endogenous IAA pools.
• IAA-amino acid conjugates have been identified as
intermediates in nondecarboxylative catabolic
pathways that deactivate IAA irreversibly
 IAA-Amino Acid Conjugates

Conjugation of IAA with amino acids leads to

irreversible deactivation. The fate of the IAA-amino
acid conjugate varies depending on the species but
usually involves oxidation of the indole ring and
subsequent addition of one or more glucose residues.
 Reversible Conjugations

IAA also exists as chemical conjugates such as glycoysl esters. These

conjugates are inactive but release free active IAA by enzymatic
hydrolysis.
These pools of IAA conjugates appear to be a source of active hormone
for the embryo during the first few days of germination. It is unknown
what role if any is played by these conjugates in mature plants.
 Gibberellins
• Gibberellins are members of a large and varied
family of plant molecules known as the terpinoids.

• Gibberellins are an extensive family of molecules

with over 125 now known, with additional members
being added.

• Gibberellins can be defined on the basis of their

chemical structure as much as by their biological
activity.
All gibberellins are diterpenes based on the
20-carbon ent-gibberellane structure
 C19-GA (GA1) C20-GA (GA37)

A little more than one third of the gibberellins characterized to date retain the
full complement of 20 carbon atoms and are known as C 20-gibberelllins. The
others have lost a carbon atom and are termed C19-gibberellins.

All gibberellins that are demonstrated to be naturally occurring and have been
chemically characterized are assigned an “A” number. This number relates to
the order of discovery of the molecule.
• GA3also known as gibberellic acid was one of the first to
be isolated and characterized.

• Because GA3 is readily extracted from fungal cultures it

is the most common commercially available form and
thus probably the most studied.

• However GA1 and GA20 (C19-GAs) are probably the most

active and therefore most important gibberellins in
higher plants.
 Why so many GA’s?
• It appears that a large proportion have little or no biological
activity.

• They are either intermediates in the synthesis of active forms or

metabolic products that still retain the basic structure but are no
longer biologically active.

• The number of gibberellins found in any one species or organ may

be very small and the number of active forms smaller yet.

• There is evidence to suggest that GA1 may be the principle if not

only active gibberellin regulating stem elongation in higher plants.
• A carboxyl group at C7 is a feature of all GAs and is
required for biological activity and C19-GAs are more
biologically active than C20-GA molecules.

• Those GAs with 3-b-hydroxylation, 3-b,13-dihydroxylation

are generally more active and those with both 3-b-OH and
1,2-unsaturation demonstrate the highest activity.
Gibberellin Biosynthesis
• The principle sites of gibberellin
biosynthesis are developing seeds and
fruits, the young leaves of developing
apical buds, elongating shoots and the
apical regions of roots.

• Gibberellins are terpenes sharing a core

pathway with several other hormones
and a wide range of secondary
products.

• Gibberellins are synthesized from the

precursor molecule isoprene (5-carbon)
via mevalonic acid (C6)
• Gibberellins are terpenes
sharing a core pathway with
several other hormones and a
wide range of secondary
products.

• Gibberellins are synthesized

from the precursor molecule
isoprene (5-carbon) via
mevalonic acid (C6).

• The products D3iPP and D2iPP

enter the terpene biosynthetic
pathway where sequential
addition of isopentyl
pyrophosphate units form
terpenes of increasing carbon
number.
• Gibberellin synthesis
from GGPP to GA12-7-
aldehyde.

• GA12-7-aldehyde is
inactive but serves as
the precursor to all
other gibberellins
• Proposed pathway for
GA biosynthesis in pea.
• The major pathway
(bold arrows) occurs in
seeds and shoots.
• The pathway shown in
light arrows occurs
only in shoots.
• The asterisk indicates
known endogenous
forms.
Antigibberllins
• The principle effect of these
anti-gibberellins is to reduce
stem elongation and thus
produce plants that are
shorter and more compact
with darker green foliage.
• Flower size in unaffected in
treated plants. These
compounds block the
synthesis of gibberellins and
their effects can be reversed
by the application of
gibberellins to the plant.
Gibberellins are deactivated by 2b-hydroxylation (Best
Characterized)
Other GA Inactivation Pathways
 Cytokinins

Cytokinins are N6-substituted derivatives of the purine base adenine and are
characterized primarily by their ability, in combination with auxin, to stimulate cell
division. In addition to stimulating cell division, cytokinins also influence shoot and
root differentiation in tissue culture, the growth of lateral buds and leaf expansion,
chloroplast development and leaf senescence.
• Kinetin (N6-furfuryl adenine) was the first
molecule discovered with cytokinin activity.
This molecule is however a synthetic cytokinin
originally isolated from autoclaved DNA. The
most widespread naturally occurring cytokinin
in higher plants is Zeatin.
 
• The major site of cytokinin biosynthesis in
higher plants is in the root. High levels have
been found in roots, in particular in the root tip.
• It is believed that cytokinins are transported to
the aerial portion of the plant through the xylem.

• High levels of cytokinins are also found in

immature seeds and developing fruits. There is
some evidence to suggest that seeds and fruits
are capable of synthesizing cytokinins it remains
equally possible that due to their high metabolic
activity and rapid growth they may simply
function as a sink for cytokinins transported from
the roots.
• IPT (adenosine phosphate-isopentyltransferase) Plants primarily use ATP or ADP
• CYP735A (CYP450 monooxygenase) upregulated by CK down regulated by IAA or ABA
• Grey arrows not well characterized at the genetic level
• Direct activation pathway catalysed by LONELY GUY (LOG) only metabolises CK nucleotides and not AMP
indicating that it is specific for CK metabolism.
• Cytokinins may be reversibly or irreversibly
deactivated by conjugation and irreversibly
deactivated by oxidation.

• Both of these methods regulate cytokinin

activity levels by the removal of cytokinins from
the active pool. Conjugations with either
glucose or amino acids have both been
demonstrated.
 Cytokinin Oxidase Degrades Cytokinin by
Removing the Side Chain

CKX catalyses the oxidation of the secondary amine group

on the side chain of the adenine ring.
The resulting imine product in nonenzymatically hydrolysed.
Aromatic CK are resistant to CKX
 Conjugations Resulting in Permanent
Deactivation of Cytokinins

N- glucosylation at the 3,7 or 9 The 9 Nitrogen is

Nitrogen of the adenine ring conjugated with alanine
 Glucosylation of the side chain hydroxyl
group

This method appears to be a mechanism for the storage of

excess cytokinins for retrieval when physiological
conditions warrant.
O-glucosides are also resistant to oxidation and thus may
serve to protect the hormone from oxidation while being
transported to a target tissue.
 Abscisic Acid (ABA)

ABA differs from the previous three classes of

plant hormones in that it is a single
compound.
 ABA is a 15 carbon sequiterpene derived from
mevalonic acid. Two possible pathways exist for
the synthesis of ABA.
 
a) direct synthesis from a 15 carbon precursor

b) indirect synthesis via cleavage of a 40 carbon

xanthophylls
 
• In fungi the direct synthesis
process occurs via
farnesylpyroposphate
intermediate.

• However in higher plants it has

been established that the
indirect synthetic pathway is
used.

• In this process ABA is

synthesized from violaxanthin
or a closely related xanthophylls
(depending on the plant
species).

• Violaxanthin is converted to
xanthoxin and then to ABA.
Early Stages of Indirect ABA Synthesis
Later Stages
• ABA appears to be synthesized in the cytosol of leaf
mesophyll cells, but due to intracellular pH gradients, ABA
accumulates in the chloroplast.

• At low pH ABA exists in its protonated form ABAH which can

freely cross cellular membranes.

• The dissociated form ABA- is impermeant as it is a charged

molecule that does not easily cross membranes.

• Thus ABAH tends to diffuse from cellular compartments with

a low pH into compartments with a higher pH. At the higher
pH ABAH dissociates and thus becomes trapped. ABA is able
to move rapidly out of the leaves to other parts of the plant.
ABA is rapidly metabolized when applied exogenously to
plant tissues. A glucose ester has been found in some plant
species but the principle metabolic route appears to be
oxidation to phaseic acid with a subsequent reduction of
the ketone group on the ring to form dihydrophaseic acid.
 Brassinosteroids
• Brassinosteroids are steroid hormones with a
chemical structure similar to the steroid
hormones in animals.

• These molecules elicit a wide range of effects

on growth and development when applied to
plants. In particular they stimulate shoot
elongation and ethylene production and inhibit
root growth and development.
 Brassinosteroids
• The first of this class to plant hormone to be identified was
brassinolide (1979). Since then more than 40 analogs have been
isolated from more than 60 plant species and virtually all tissues
including pollen, seeds, stems, roots and flowers.

• Brassinolide is the most biologically active brassinosteroid and is

widely distributed throughout the plant kingdom. It is present at
very low levels however like other hormones it is active at
micromolar concentrations.

• For many years the classification of brassinosteroids as hormones

was not widely accepted, however it is now accepted that they play
an important role in normal plant development.
Structure of Brassinolide and Casasterone
An Overview of Brassinosteroid Biosynthesis
• Several deactivation mechanisms of brassinosteroids have been
reveled through studies in a number of plant species. The a-hydroxyl
groups on the A ring may be epimerized to form a b-hydroxyl, which is
then followed by esterfication with fatty acids or by glucosylation.

• Other possibilities include the cleavage of the side chain or conjugation

at other hydroxyl positions. The precise method of deactivation
depends on the species of plant and possibly the tissue involved.
 Ethylene
• Ethylene is a simple gaseous hydrocarbon with the
chemical structure H2C=CH2.

• Ethylene is not required for normal vegetative growth,

although it can play a significant role in the
development of roots and shoots.

• It appears to be primarily synthesized in response to

stress and produced in large amounts by tissues
undergoing senescence or ripening.
 Ethylene
• Ethylene is often produced when high concentrations
of auxins are applied to plant tissues.

• Ethylene occurs in all plant organs although the rate of

production may vary depending on the stage of
development.

• Ethylene production can also vary from tissue to tissue

within the organ and is frequently located in peripheral
tissue.
• Ethylene production is promoted by a number
of factors including IAA, wounding and water
stress primarily through the induction of the
synthesis of ACC synthase.

• Ethylene synthesis is also stimulated by

ethylene itself, a form of autocatalysis
commonly seen in fruit ripening.
• Unlike other plant hormones, ethylene is a volatile
gas that is readily given off to the atmosphere.

• However ethylene can be metabolized by oxidation

to CO2 or by conversion to ethylene oxide or
ethylene glycol.

• It is still unclear whether such reactions play a role

in the regulation of the levels of ethylene, but it is
most likely that tissues lose excess ethylene by
simple diffusion into the surrounding atmosphere.