FISIOLOGIA VEGETAL COMPLEMENTAR

(2013/2014
Phytohormones and Elicitor Molecules
CYTOKININS

Cytokinins participate in the regulation of many
plant processes, including cell proliferation,
morphogenesis of shoots and roots, nutrient
acquisition, vascular development, light
responses, and senescence.




In the intact plant, cells can be
stimulated to divide by wounding,
infection, and plant hormones, including
cytokinins (Figure 21.1).
Cytokinins are N-substituted adenine derivatives that initiate cell proliferation in
many plant cells in the presence of auxin.
Cell Division and Plant Development
Kinetin is not a naturally occurring plant growth regulator. It is a by-
product of the heated-induced degradation of DNA, in which the
deoxyribose sugar of adenosine is converted to a furfuryl ring and
shifted from the 9 pisition to the 6 position on the adenine ring.
The Discovery, Identification and Properties of Cytokinins

Zeatin is the most abundant naturaly occurring free
cytokinin, but dihydrozeatin (DHZ) and isopentenyl
adenine (iP) also are found (Figure 21.2).
Nat Nat
Nat
Nat
Nat
Nat
The cytokinins can be present in the plant as
ribosides (in which a riboside sugar is attached to
9 nitrogen of the purin ring) ribotides (in which
the ribose sugar moiety contains phosphate
group) glycosides (in which a sugar molecule is
attached to the 3, 7, or 9 nitrogen of the purine
ring or to the oxygen of the zeatin or dihydrozeatin
side chain), or in other conjugated forms.
The Discovery, Identification and Properties of Cytokinins

Some synthetic compounds can mimic cytokinin action


Cytokinins are defined as compounds that have biological activities similar to
those of trans-zeatin. These activities include:

Inducing cell division in callus cells in the presence of an auxin

Promoting bud or root formation from callus cultures when in the
appropriate molar ratios to auxin

Delaying senescence of leaves

Promotion expansion of dicot cotyledons
The Discovery, Identification and Properties of Cytokinins

Some plant pathogenic bacteria, fungi, insects, and nematodes secrete active
cytokinins, / or cause the plant cells to synthesize plant hormones, including
cytokinins, which in some cases induce abnormal growths (Figure 21.3). The
cytokinins produced by microorganisms include trans-zeatin, iP, cis-zeatin, and
their ribosides, as well as 2-methylthioderivatives of zeatin.
For example, increased cytokinin, supplied by
interacting bacteria, fungi, viruses, or insects, can
cause an increase in the proliferation of the shoot
apical meristem and / or the growth of lateral buds,
which normaly remain dorment. This proliferation is
known as fasciation, as it is the case of the so called
witches broom (Figure 21.3).

Certain insects secrete cytokinins, whic play a role in
the formation of galls the insects use as feeding sites.

Root-knot nematodes also produce cytokinins, which
may be involved in manipulating host development to
produce the giant cells, from which the nematodes
feed.
Biosynthesis, Metabolism, and Transport of Cytokinins
Cytokinins are synthesized in roots, developing embryos, young leaves, fruits, and crown
gall cells tissues, as well as by plant-associated bacteria, fungi, insects, and nematodes.

From infection with Ti plasmid, crown gall cells have acquired T-DNA carrying genes for the
biosynthesis of cytokinins, auxin, and opines (Figure 21.4).
Biosynthesis, Metabolism,
and Transport of Cytokinins
The side chains of the cytokinins contains one
isoprene unit.
The precursor for the formation of these
isoprene structures is dimethylllalyl
diphosphate (DMAPP), which is derived from
either the mevalonete pathway (primarily for
cis-zeatin) or the methylerythriol phosphate
(MEP) pathway (primarily for DHZ, iP, and
trans-zeatin).
The product of IPT gene (isopentenyl
transferase IPT) catalyzes the first commited
step in cytokinin biosynthesis (Figure 21.5).

Cytokinins are passively transported through
the xylem and phloem
Cytokinins are rapidly metabolized by plant tissues

Cytokinin oxidase irreversibly inactivates cytokinins, cleaving the side chain fom
zeatin (both cis and trans), zeatin riboside, iP, and their N-glucosides, contributing
to their regulation. However, dihydrozeatin and its conjugates, as well as aromatic
cytokinins such as benzyladenine, are resistant to cleavage.

N-conjugations are generally irreversible. The O-conjugations are reversible, as glucosidase
enzymes can remove the conjugates from the side chain.

The level of active cytokinin results from positive and negative influences on their
synthesis, conjugation, and transport.

Biosynthesis, Metabolism, and Transport of Cytokinins
Cytokinin biosynthesis in plants occurs primarily in plastids
The Biological Roles of Cytokinins
Cytokinins promote shoot
growth by increasing cell
proliferation in the shoot apical
meristem.

Reducing endogenous cytokinin
by overexpression of cytokinin
oxidase or mutation of the IPT
gene severely limits shoot
growth and produces a smaller
shoot apical meristem (Figures
21.10, 21.11).
Mutation of all thee cytokinin
receptors in Arabidopsis eliminates
perception of cytokinin and results
in multiple developmental defects
including a reduced shoot apical
meristem, a stunted shoot, and
greatly reduced flowering (Figure
21.12).
The biological Roles of Cytokinins
Cytokinins interact with other hormones and with key transcription factors involved in regulating
shoot apical meristem function.

In the root apical meristem, auxin promotes cell division and cytokinin promotes cell
differentiation. In contrast with their action in shoots, cytokinins inhibit root growth by
promoting the exit of cells from the root apical meristem.

In contrast to its effect on the shoot, overexpression of cytokinin oxidase in tobacco increases
root growth, primarily by increasing the sise of the root apical meristem (Figures 21.13, 21.14).
The biological Roles of Cytokinins
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The Biological Roles of Cytokinins
Cytokinins regulate specific components of the cell cycle

Cytokinins regulate cell division by affecting the controls that govern the passage of the cell
through the cell division cycle.

Zeatin levels peak in synchronized culture tobacco cells at the end of S phase, the G2/M
phase transition, and in late G1.

Inhibition of cytokinin biosynthesis blocks cell division, and application of exogenous
cytokinin allows cell division to proceed.

Both auxin and cytokinins participate in regulating the cell cycle by controlling the activity
of the cyclin-dependent protein kinases (CDKs).

Auxin regulates the expression of the gene that encodes the major CDK, Cdc2 (cell
division cycle 2).

Cytokinin has been linked to the activation of a Cdc25 like phosphate, whose role
is to remove an inhibitory phosphate group from the Cdc2 kinase. This action of
cytokinin provides one potential link between cytokinin and auxin in the cell
cycle: regulating the passage from G2 to M phase.
In the root apical meristem, auxin promotes cell division and cytokinin promotes cell
differentiation.

Both cytokinin and auxin regulate the plant cell cycle and are needed for cell divison.

Cytokinins elevate the expression of the CYCD3 gene, which encodes a D-type cyclin.
Overexpression of CYCD3 gene can bypass the cytokinin requirement for cell
proliferation in culture. (Figures 21.15).
The Biological Roles of Cytokinins

The auxin:cytokinin ration regulates morphogenesis in cultured tissues.

Whereas high auxin : cytokinin ratios stimulated the formation of roots, low auxin :
cytokinin ratios lead to formation of shoots. At intermediate levels , the tissue grew
as an undifferentiated callus (Figure 21.16).
The Biological Roles of Cytokinins

The ration auxin / cytokinin
determinates the differentiation of
cultured plant tissues in either
roots or shoots.


Mutating the ipt gene (the tmr
locus) of the Ti plasmid blocks
zeatin biosynthesis in the infected
cells. The resulting high auxin :
cytokinin ratio in the tumor cells
causes the proliferation of roots
instead of undifferentiated callus
tissue. In contrast, mutating,
either of the genes for auxin
biosynthesis (tms locus) lowers the
auxin : cytokinin ratio and
stimulates the proliferation of
shoots (Figure 21.17).
The Biological Roles of Cytokinins
Cytokinins modify apical dominance
and promote lateral bud growth.

Auxin maintains apical dominance by
repressing the local synthesis of cytokinin in
lateral buds and increasing the expression
of cytokinin oxidase (Figure 21.18).
The Biological Roles of
Cytokinins
Cytokinins delay leaf senescence (Figure 21.19)
and promote nutrient mobilization (Figure 21.20)
The Biological Roles of Cytokinins
Cytokinins help regulate the synthesis of photosynthetic pigments and
proteins (Figure 21.21) and regulate vascular development.
The Biological Roles of Cytokinins
Photosynthetic productivity may be increased by making cytokinin-overproducing
plants that have delayed senescence or yield more grains (Figures 21.22, 21.23).
The Biological Roles of Cytokinins
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