Gibberellins

Outline

 Discovery of gibberellins (GAs)

 Green Revolution (late 1960s)
 General introduction
 Biosynthesis
 Homeostasis
 Signaling pathways
 Physiological functions
 Gibberellins were identified as
fungal compounds that promote stem
elongation
Bakanae disease Bakanae disease means
(Gibberella fujikuroi) “foolish seedling”, because
infected plants elongate too
rapidly, and are unable to
Uninfected support themselves; they
plant are also male sterile.

Infected,
hyper-
elongated
plants

Gibberellin A3 (GA3)

Tetracyclic diterpenoid phytohormone

Later, gibberellins were identified as
endogenous plant growth regulators
Gibberellins promote stem
elongation and seed
germination. In some plants
Wild-type pea
they promote flowering and fruit
development.

Mendel’s short peas

are deficient in GA
synthesis.

Some plants are treated with GA- Development

synthesis inhibitors to maintain a of many
shorter stature and prevent seedless fruits
lodging (tipping over). requires GA
application

Lester et al. 1997. The Plant Cell 9, 1435-1443

 The manipulation of GA levels is
tremendously important for agriculture
One of the most significant
accomplishments of 20th
century science was the
development of semi-dwarf
grain varieties which are
deficient in GA synthesis or
response.

Received Noble peace prize for

contributing to world peace through
increasing food supply (1970)

Distinguished plant breeder: Prof. Norman Borlaug (1914-2009)

 GA has been described as “an
inhibitor of an inhibitor”

A car doesn’t roll down a

hill when the brake is on
to inhibit it. Releasing the
brake “inhibits the
inhibitor”, allowing
another force (gravity) to
move it.

GA doesn’t promote
growth on its own, but
instead inhibits growth
inhibitors (i.e. releases
the brakes).
GA synthesis and homeostasis

In 1956, B.O. Phinney

found that some dwarf
mutants of Zea mays
could be rescued by
GA application.

The pathway of GA biosynthesis

in plants was determined in part
by analysis of GA-deficient
Wild type Wild type dwarf-1 dwarf-1 dwarfs; an early use of the
+ GA + GA
“chemical genetics” approach.

Phinney, B.O. 1956. PNAS 42, 185-189

GA biosynthesis occurs at four principle sites:

1. Flowers (especially anthers)

2. Developing seed and fruits; germinating seeds
3. Young leaves
4. Root tip

Bioactive GAs:

GA1, GA3, GA4, GA7 (shoots)

GA54, GA55, GA60, GA61 and GA62 (cereal grains)

Vegetative tissues may have different GAs than
reproductive tissues:

e.g. Wheat plant (GAs in vegetative tissues responsible for shoot growth
stimulation are GA1, GA3, GA4 and GA7; whereas additionally GAs such as
GA54, GA55, GA60, GA61 and GA62, which are not present in vegetative
tissues, are found in developing grains.

Functions of GAs:
Stimulation of organ expansion and of certain developmental transitions,
such as seed germination, flowering and between the juvenile and adult
phases
 The GA biosynthetic pathway is complex

GA13 hydroxylase

CPS= ent-copalyl diphosphate synthase; KS= ent-ent-kaurene synthase; KO= ent-kaurene oxidase;
KAO= Kaurenoic acid oxidase; GA20ox= GA20oxidase; GA3ox= GA3oxidase; GA2ox= GA2oxidase
 Loss-of-function mutants of CPS or
KS are severely dwarfed

GGPP CPP ent-

kaurene Wild-type and dwarf
CPS KS
mutant rice

These early enzymes are

encoded by one or at most
few genes, so mutants have
severely reduced GA levels. oscps-1 osks-1

WT oscps-1 osks-1

Sakamoto et al. 004. Plant Physiol. 134, 1642-1653

Similarly, loss-of-function mutants of
KO or KAO are severely dwarfed
The pea
gene LH
encodes ent-
The pea gene NA kaurene
encodes ent- oxidase
kaurenoic acid (KO)
oxidase (KAO)
ent-
kaurene
KO

ent-kaureonic
acid
KAO
GA12

Davidson et al. 2003. Plant Physiol. 131, 335-344

Davidson et al. 2004. Plant Physiol. 134, 1123-1134
 The later enzymes are encoded by
multiple genes with differing
expression patterns
GA 13-
hydroxylase
GA12 GA53
GA 20-
The “green revolution” oxidase
semidwarf1 rice variety
is mutated in a GA20ox GA9 GA20
GA 3-oxidase
that is expressed in
shoots but not GA4 Active GA1
GAs
reproductive tissues,
leading to increased
grain yields.

Sasaki et al. 2002. Nature 416 (6882), 701-702

 Similarly, mutants in GA3ox genes
can have reduced height but not
seed production
Pea Arabidopsis
GA 13-
hydroxylase
GA12 GA53
GA 20-
oxidase
The le mutant
that Mendel GA9 GA20
WT studied is GA 3-oxidase
mutated in a (le)
WT le ga4 GA3-oxidase-
GA4 (ga4) GA1
encoding gene,
as is the ga4
mutant of
Arabidopsis.

Lester et al. 1997. The Plant Cell 9, 1435-1443

Chiang et al. 1995. The Plant Cell 7, 195-201
GAs can be deactivated by several
different enzymes

HO
GA4 GA2ox
HO HO

Elongated
Uppermost
GA Methyl- Internode
Transferase (EUI)
(GAMT)

HO

HO
CH3
Dwarfism is conferred by overexpression of the
GA deactivating enzyme GA 2-oxidase
GA 13-
Arabidopsis
Arabidopsis hydroxylase
GA 2-
GA12 GA53 GA 2-
oxidase oxidase
GA 20-
oxidase
Inactive Inactive
forms GA9 GA20 forms
GA 3-oxidase

GA4 GA1

GA 2-oxidase
WT
WT GA2ox GA2ox
GA2ox GA2ox Inactive Inactive
Tobacco forms forms

Different GA2oxs can preferentially

act on active GAs or their inactive
WT GA2ox GA2ox precursors, with different
developmental outcomes.

Schomburg et al. 2003. The Plant Cell 15, 151-163

ELONGATED UPPERMOST INTERNODE
encodes a GA-deactivating enzyme
Mutation Mutation Mutation Over-expression

Loss-of-function mutants
are elongated, and
overexpression lines are
dwarfed.

Zhu et al. 2006. The Plant Cell 18, 442-456

 Overexpression of a GA-
methyltransferase causes dwarfism

GAMT1
GAMT1
Tobacco

WT

WT

Petunia

Varbanova et al. 2007. The Plant Cell 19, 32-45

 GA biosynthesis and deactivation
are tightly controlled

Temperature
and light Active GAs
Auxin upregulates regulate downregulate GA
GA synthesis. GA3ox synthesis and
Auxin upregulate GA
deactivation.

Most genes are

expressed in a
cell-specific
manner

Thomas, S.G., Phillips, A.L., Hedden, P. 1999. PNAS, 4698-4703

 Paclobutrazol inhibits GA synthesis

0 M PAC 1 M PAC 5 M PAC 25 M PAC

Paclobutrazol is often sprayed on

plants to interfere with GA
synthesis and prevent branch
growth (e.g. in fruit trees).

Arabidopsis plants treated with increasing

amounts of paclobutrazol (PAC).

Rieu et al. 2008. The Plant Cell 20, 2420-2436

Tissue-specific hormone modulation
can optimize growth and crop yields

ACTPRO::GA2OX
Increasing GA catabolism
everywhere using a
constitutive promoter
affects seed production.

GA3OXPRO::GA2OX
Increasing GA
catabolism only in
internodes produces
high-yielding dwarf
rice plants.

Hedden, P. 2003. Constructing dwarf rice. Nature Biotech. 21, 873-874

Unlike auxin, GA is not transported
in a polar way

The same amount of GA

moves from the upper
donor block to the lower
Normal block no matter the
orientation polarity of the stem
segment. By contrast,
auxin moves much more
efficiently from stem apex
to base.
Inverted
orientation

Kato, J. 1958. Science 128, 1008-1009

 GAs are graft-transmissible; they
can move long distances

WT - d1

In pea, a mutant
na shoot is
rescued by
grafting onto a Maize seedlings are d1- d1
Na root. grafted side-by-side

na In maize, GA or a GA-
Na precursor moves from the
na wild-type plant to d1 and
na promotes growth.

Proebsting et al. 1992. Plant Physiol. 100, 1354-1360

Katsumi et al. 1983. Plant Cell Physiol. 24, 379-388
 Perception and signaling

DELLAs GA

GA inactivates DELLA inhibitors

Without the inhibitory DELLAs, the

plant can respond to growth-
promoting activities.
GA responses

Biosynthesis mutant Response mutant

No GA signaling

Harberd et al. 2009. Plant Cell 21, 1328-1339

 GA receptor discovery

1. Early, microinjection experiments with membrane

impermeable GAs using isolated aleurone protoplasts
from barley grains
Hooley et al., 1991, 1992; Beale et al., 1992; Gilroy and Jones, 1994

- GA response was induced when protoplasts incubated with the GA

solution, but not when injected into the protoplast, suggesting the
presence of plasma membrane bound GA receptor

2. Interesting study by Ueguchi-Tanaka et al. (2005) in

rice has provided a strong evidence for the nuclear
based GA receptor, known as GA-INSENSITIVE DWARF1
(GID1)
The major components involved in GA signaling
are GID1 receptor, DELLA and SCF complex.

GID1 (GA receptor)

It is localized in the nucleus (Ueguchi-Tanaka et al., 2005). Binding of the GID1

receptor to GA molecule causes a conformational change in the GID1
allowing it to interact with DELLA proteins. This activated GA-GID1-DELLA
complex has an enhanced affinity for the SCF complex.
DELLA (growth repressor)

- Each DELLA protein contains two major domains

1st domain: N-terminal DELLA and TVHYNP motifs
2nd domain: C-terminal GRAS domain

- The DELLA domain has a high affinity for the GID1 in a GA dependent
manner, whereas the GRAS domain has a role in the interaction of
DELLA with F-box protein of the SCF complex

SCF complex

Is an E3 ligase that consists of skp1, cullin, F-box protein and RING-H2

motif. The binding of the F-box protein to the activated GA-GID1-DELLA
complex induces the addition of ubiquitin to the DELLA. This process of
tagging the DELLA with ubiquitin is known as the ubiquitination process,
which later targets the DELLA for degradation by the 26S proteasome
With GA
Without GA
Proposed model for the interaction
between the light and GA signaling
during seed germination in Arabidopsis

a. in the dark and under GA depleted condition,

b. In the dark and in the presence of GA (normal situation)

c. in the presence of the light and GA

DELLA-encoding genes have great
agricultural impact

The green-revolution gene

reduced height1 from wheat
encodes a mutant DELLA.

Wild-type wheat

Reduced-height1 (Rht1) wheat

Peng et al. 1999. Nature 400, 256-261

Comparison of GA signaling pathways
Antagonism of ABA and GA, the regulation of GA biosynthesis by DELLA
Roles of GAs in whole-plant physiology
• GA stimulates cell division and expansion to control growth and
elongation in plants

• GA mediates stress responses through DELLA proteins

Cold or salt stress or flooding induces ethylene, which induces DELLA
slowing down the plant growth

• GA brings about reserve mobilization, thus promotes seed

germination

• GA promotes flowering in all plants, but inhibits tuberization in

potato

• GA application suppresses leaf senescence, whereas GA inhibitors

like paclobutrazol accelerates; possibly by antagonizing the effects of ABA
GA acts antagonistic to ABA during seed
germination

ABA promotes Germination

desiccation tolerance
and dormancy

Seed germination requires Reserve

mobilization
elimination of ABA and
production of GA to
promote growth and Cell
expansion
breakdown of seed storage
products.

GA
ABA
The function of GAs during seed germination

Seed coat

Endosperm starch
Starchy endosperm

Hydrolysed sugars
Active α-amylase

α-amylase m-RNA
Cotyledon

α-amylase gene
Epicotyl
Aleurone cells layer

Growth stimulated GA molecules diffused to the endosperm initiate α-

amylase synthesis by aleurone cells layer
Radicle
Imbibition in water produces GA
molecules in the embryo or scutellum
 GAs regulate tuber formation in potato
Reduced GA levels stimulate stolon-to-tuber transition in potato

Kondhare et al. 2021. Plant Physiol. 187(3), 1071-1086

GAs role in tuberization pathway

Kondhare et al. 2021. Plant Physiol. 187(3), 1071-1086

 GAs role in parthenocarpy (seedless
fruits)
Reported in citrus fruits, strawberry, tomato, banana, watermelon, pears, kiwifruit etc.

Joldersma and Liu, 2018 J. Exp. Bot. 69(5), 955-962

Emerging mechanisms for fruit setting and parthenocarpy

 Epigenetics: DNA methylation and PRC2 mediated histone modifications

 Certain MADS-box genes: PISTILLATA (PI); DEFICIENS (DEF) and AGL6

Joldersma and Liu, 2018 J. Exp. Bot. 69(5), 955-962