Gibberellins

• Gibberellins are usually associated with plant height. If you apply them to a dwarf plant, the
plant will grow very high. It can overcome genetic dwarfism. While a normal plant treated
with gibberellins shows no response. It can also induce flowering in plants.

• Discovery: In Japan, rice is an economically important plant. Towards the end of the 19th
century, a disease affected the rice plants and resulted in loss of almost 40% of the crop. The
disease was called bakanae or foolish seedling. The plant grows exclusively high that they
cannot support themselves. They become sterile (no seeds), and chlorosis takes place
(yellowing-plants/pale → no chlorophyll produced) and eventually die. They started doing
research and they managed to find a relation between this disease and a fungus called
Gibberella fujikuroi. Later, the scientists in the West became interested in studying this
disease, and they noted that we don’t need the living fungus to infect. They managed to
extract organic chemical from the fungus, and this chemical, when applied to a normal plant,
was enough to cause the same symptoms. They called these chemicals gibberellins.

• Gibberellins has a carboxyl group → Gibberellic acid (GA). There are more than 100 types of
GA, numbered in the order of their discovery. They were discovered in almost all plants but
never in bacteria. These GA have a complicated structure that might remind you of
cholesterol. 4 rings fused together (either 19 or 20 Carbon atoms). They are lipids derived
from isoprene (5 Carbons).

• The synthesis of GA is very similar to cholesterol synthesis, and this same pathway is shared
with carotenoids, ABA, steroids, plastoquinone, and cytokinins (they all start with Acetyl-CoA
precursor and an IPP intermediate):
Acetyl-CoA →→→ Mevalonate →→→ IPP (isopentylpyrophosphate)

• GA is synthesized either in cytosol (Mevalonate pathway), or in plastid/chloroplast

(Mevalonate independent pathway).

• Mevalonate-dependent Pathway:

- Acetyl-CoA + Acetyl-CoA → Acetylacetyl-CoA + Acetyl-CoA → hydroxymethylgulataryl-

CoA (HMGCoA) → Mevalonic acid (6C / key acid in the synthesis of biologically important
molecules)
 - Mevalonate → Mevalonate Pyrophosphate → IPP (5C / through decarboxylation).
This IPP is an important precursor (it is an isoprenoid) that will lead to the synthesis of
many important compounds.
- IPP → Δ2-IPP (dimethyl allyl pyrophosphate / isomer / change position of double bond)
- IPP + Δ2-IPP → Geranyl-PP (10C) + IPP → Farnesy-PP (15C) + IPP → Geranylgeraniol-
PP (GGPP / 20C)
- GGPP will undergo cyclization, and this occurs in the plastids.
- GGPP → Copalyl diphosphate (first cyclic compound / first intermediate containing a
ring structure) → Kaurene (4 ring structure) → Kaurenoic acid (product of oxidation of
Kaurene in ER) → GA12-Aldehyde (precursor of all other Gibberellins) → GA12-carboxylic
acid (by oxidation) → series of oxidations in the cytosol leading to gibberellins.
- Synthesis starts in cytosol, but also the endoplasmic reticulum and plastid are involved.
- GA can either be 19C or 20C.
- Some gibberellins are more active than others.

• Mevalonate-independent Pathway:
- Starts in the plastids.
- It involves G3P and pyruvate.
- No mevalonate intermediate

• The synthesis of GA usually occurs in the embryos of the seeds, in the end leaves, in the roots,
and in intercalary meristems of upper internodes.
• Once GAs are synthesized, they are transported. The transport of GA is not polar. It occurs in
sieve tubes of the phloem. They control physiological activities when they reach target cells
and then they will be inactivated.

• Inactivation of GA:
- Adding OH at a key position → structure modification (example: adding a hydroxyl group
to GA4 converts it to GA34 which is much less active)
- Formation of conjugates → stored form of GAs by attaching them to a molecule. These
molecules are usually small metabolizes (glucose) to form an ester bond or an ether bond.
Whatever is the bond, gibberellin glucoside will be formed and this is a stored form.

• Synthetic compounds:
- There are many synthetic compounds that act as anti-gibberellins or growth retardants.
They inhibit key enzymes in the synthesis process, preventing the synthesis of
gibberellins.
 - Example: AMO1618, Phosphon D, and Cycocel (CCC) prevent the conversion of GGPP to
form CPP (the first cyclic step) by inhibiting the enzyme CPS (copalyl pyrophosphate
synthase) → the production of the GA will be blocked.
Some of them even act on more enzymes (phosphon D can act on the next step that
converts CPP to kaurene → it inhibits Kaurene synthase enzyme).
The chemistry of these compounds have been identified, but these are commercial
names.

• Physiological Effects of GA:

1. Promote growth:
(like auxins). If you have a dwarf pea and apply GA, it will grow to its normal height.
Such plants are dwarf because the synthesis of endogenous GA is inhibited.
Example corn d5 is a mutant that grows only 1/5 of its normal height. Corn d5 lacks the
enzyme Kaurene Synthase, and hence it cannot convert CPP to Kaurene and GA cannot
be produced. After adding GA, normal and dwarf plants will have the same phenotype
but different genotypes. This is called phenocopy.
However, sometimes you might add GA to a dwarf plant and it will not show any effect.
This is because they lack the receptors of GA. This may be due to ABA that acts as inhibitor
of growth. So this plant has an excess of ABA that counteracts the effects of GA. You could
also have other naturally occurring inhibitors.

2. Cell elongation through XET:

Xyloglucan Endotransglucosylase enzymes cut a part of the xyloglucan and add it to the
non-reducing end of another xyloglucan. This will give more room for expansins to act.
Growth of a plant depends on both auxins and GA.

3. Cell cycle regulation:

In intercalary meristems (at base of upper internodes), GA regulates the transcription of
cycle-dependent kinases (CDKs) which induce the transition from G1 stage to S → cell
cycle activation and stimulating division→ more expansion and growth.
 GA affects the stem and not the root. Sometimes, some dwarf plants show response to
GA in both the roots and stems, but this is not clear (direct vs. indirect effect).

4. Flowering:
- For the GA to have effect on flowering, two conditions must be met: Light photoperiod
and convenient temperature (cold?) (though some plants show no response to both).
These two factors are needed for gene expression and transcription, which will lead to
the formation of GA and other hormones.
Based on light:
- Long day plants (LDP): the plant flowers when the light period exceeds a certain critical
length. (ex: 16 continuous hours of light). They require long days.
- Short day plants (SDP): The plant will flower if the length of the day is shorter than a
critical length.
- Neutral day plants (NDP): ex: tomato; it flowers when it reaches a certain number of
internodes (no light needed).
Actually, the most important factor in photoperiod is dark rather than the light.

- In the plant, there is a protein pigment called phytochrome. This phytochrome is present
in 2 interconvertible forms:
Pr (absorbs light in the red region 650-680 nm).
Pfr (absorbs light in the far red region 710-740 nm).
When Pr absorbs light in the red region, it will be converted to Pfr →cascade → production
of GA → the plant will flower.
In dark, Pfr is reverted back to Pr → dark reversion → dark periods limit the amount of Pfr
present.
(Pr has a tetra-pyrrole structure (open pyrrole rings/ heme group opened))

- Some plants require cold treatment to flower (vernalization) like carrots and apples.
Ex: Carrots are biennial plants that require 2 growing seasons: vegetative growth and
flowering growth.

- Plants that require long days or vernalization can be induced to flower by adding GA, since
their flowering necessitates the buildup of GA.

- GA are also known to determine sex of flower (whether flower is male or female). Recall
that the flower is made up of 4 floral parts: sepals-petals-stamen-pistil.
Most flowers are hermaphrodite (perfect flowers), having both male and female organs.
Some species, on the other hand, are imperfect (unisexual). Such flowers could be carried
 on the same plant (monoecious-cucumber), while others are dioecious (separate plants-
spinach). So GA are known to determine sex. It is known to induce stamenate flowers and
inhibit pistilate flowers from forming.
Note that sex determination can be carried out by other hormones also (ethylene).

5. Bud and seed dormancy:

Water and oxygen are necessary for a seed to germinate. However, sometimes it doesn’t
even in the presence of these two factors. This id due to the presence of large amounts
of inhibitors → seed is dormant
Also, lateral buds in leaves are dormant in winter.
How to overcome dormancy?
Three environmental factors are needed:
- Red light (Pr → Pfr )
- Certain temperature (usually low)
- Long days
GA can replace these factors and break doemancy.

6. Parthenocarpy:
It is the production of seedless fruits. Some plants can produce parthenocarpy fruits by
application of GA (ex: pome and stone fruits/ peaches and other fruits that have a stone
seed)
Apples are not responsive to auxins (only GA). Other fruits might be responsive to both
or none.
When two hormones are involved, one hormone usually induces an enzyme that leads to
the production of another hormone.

7. Mobilization of storage compounds (nutrients) during seed germination:

- When a seed germinates, it cannot do photosynthesis (no chloroplasts). So how does it
grow? In monocots, the food is stored in the endosperm, while in dicots, it is stored in the
cotyledons. They are stored as polymers → hydrolytic enzymes needed for mobilization.
- For the seed to germinate, it mainly needs water and oxygen as long as there are no
inhibiting factors.
- Structure of a barley grain (monocot):
Embryo is the part that germinates. It gives the radical (1st part to penetrate the soil, and
gives rise to the root).
Plumule gives rise to the stem including the leaves, or the shoot. This plumule (or
embryonic shoot) is surrounded by a sheath called the coleoptile.
The cotyledon in monocots is called scutellum.
The major part of the seed is the endosperm. It is the tissue where food is stored as
polymers.
Surrounding the endosperm, we have the aleurone layer. Then we have the seed coat
(testa or pericarp).
- In barley and rice, most of food is in the form of starch.

- Starch is a large molecule needed for germination. It should be broken into glucose and
mobilized. We need amylase (hydrolytic enzyme) to do so. Amylase (and other hydrolytic
enzymes) are produced in the aleurone layer de novo (new enzymes) and will have to
move to the endosperm and hydrolyze the starch so that glucose can move to the
embryo.

- GA are needed for the expression of the genes needed for the synthesis of amylase. GA
is usually present in the embryo and will move to the aleurone layer.
- How can we prove this?
If we remove the embryo. The aleurone layer can no longer produce amylase.
If we add GA → Amylase (and hydrolytic enzymes are expressed).

- Hydrolytic enzymes can be also synthesized and secreted in the part of scutellum facing
the endosperm (it is rich in both ER and GA, which concentrates secretory substances).

- How does GA affect the production of these enzyme?

In aleurone cell: GA → bind to a receptor on plasma membrane → G-protein activated →
signal transduction pathway, can be:
 Calcium-independent pathway: cGMP is secondary messenger → activates a signaling
intermediate: F-box protein → inhibits the repression of GAMYB gene→ GAMYB mRNA is
formed → ribosomes → GAMYB transcription factors → Promoter of α-amylase gene →
α-amylase mRNA → translation in ribosomes of rough endoplasmic reticulum → Golgi
apparatus → leaves in membrane vesicles that fuse with the membrane → secreted in
endosperm.
Calcium-dependent pathway: Calmodulin is the secondary messenger, activated by
calcium → Protein kinase → release α-amylase into the endosperm.

- As GA↗ → [Ca2+] ↗ and GAMYB ↗ → hydrolytic enzymes production increase.

• Commercial applications of GA:

1. Grapes:
Production of seedless grapes and the increase of the number of grapes in a cluster
(almost double). The grapes are also more loose (not packed) and less susceptible to
infection by fungal diseases.
2. Barley:
Role in germination of barley seeds. GA increases the amount of amylase: enhanced
digestion of starch to produce malt. Note that malt is a mixture of starch hydrolysis
products. This malt is used in the barley industry to produce beer. More malt → more
beer.
3. Sugarcane:
GA increases the length of sugarcane stem. However, this is not accompanied by increase
or decrease in sugar concentration.
4. Promalin:
A synthetic compound made up of a mixture of: GA4, GA7, and 6-benzyladenine
(cytokinin). This is applied to apples and increases their size.