‘BIOSYNTHESIS PATHWAYS AND

METABOLISM OF CYTOKININ’

SUBMITTED TO-
Dr.Rajendra Kumar Panda
Associate Professor,
Department of Plant
Physiology,OUAT,Bhubaneswar
 CYTOKININ-
 Cytokinin is one of the natural phyto hormones that have effects on many
physiological and developmental processes, including leaf senescence,
nutrient mobilization, apical dominance, the formation and activity of
shoot apical meristems, floral development, the breaking of bud dormancy,
and seed germination.
 It was first discovered by Folke Skoog and colleagues in the 1950s.
 “Kinetin” was the first substance to be identified as a cytokinin, and
although it is not naturally produced and has not been found in living
plants.It is a breakdown product of DNA.
 The naturally occurring cytokinin ’’zeatin(Trans-zeatin)’’ was first isolated
from immature maize endosperm in the early 1960s.It is the most abundant
natural cytokinin.
 BIOSYNTHESIS-
 The primary site of the biosynthesis of cytokinin is root tip.
 Twobiosynthetic pathways have been proposed for
cytokinin biosynthesis.
 Thefirst is the direct pathway, involving formation of N6 -
isopentenyladenosine monophosphate (i6 AMP) from AMP
and dimethylallyl pyrophosphate (DMAPP), followed by
hydroxylation of the side-chain to form zeatin-type
compounds.
 Thesecond pathway is the indirect pathway, in which
cytokinins are released by turnover of tRNA containing cis-
zeatin.
 It may use ATP, ADP, or AMP as substrates and may use dimethylalyll
pyrophosphate (DMAPP) or hydroxymethylbutenyl pyrophosphate (HMBPP)
as prenyl donors. This reaction is the rate-limiting step in
cytokinin biosynthesis. DMAPP and HMBDP used in cytokinin biosynthesis
are produced by the methylerythritol phosphate pathway (MEP)
 Cytokinin can also be produced by recycled tRNAs in plants
and bacteria. tRNAs with anticodons that start with a uridine and
carrying an already-prenylated adenosine adjacent to the anticodon
release on degradation the adenosine as a cytokinin.
 The prenylation of these adenines is carried out by tRNA-
isopentenyltransferase.
 Auxin is known to regulate the biosynthesis of cytokinin.
1.iPMP dependent Pathway-
 The precursor for the cytokinin is adenosine.Along with that ADP/ATP in
order to produce the intermediate which will help the plant cell to
produce zeatin.
 If adenosine will react with ADP/ATP then Dimethylallyl pyrophosphate
(DMAPP) will be formed.Again if ADP will be associated with DMAPP in the
presence of enzyme isopentyl Transferase(IPT) ,it will form isopentyl
adenosine -5-diphosphate(iPDP).
 If ATP is associated with DMAPP in the presence of same enzyme
then,isopentyl adenosine-5-triphosphate(iPTP) will be produced.
 Both iPDP and iPTP will be converted into Zeatin(Cis-trans isomer
conjugate) the presence of the enzyme hydrolase .
2.iPMP independent pathway-
 Plant genes with considerable homology to these microbial genes
encode tRNA isopentenyltransferases, which mediate the transfer of an
isopentenyl group to adenine of tRNAs recognizing the codon group.
 The indirect pathway involves release of cytokinins from breakdown of
such tRNA. Although the weakly active ciszeatin is the major cytokinin
in plant tRNA.
 cis-zeatin can be converted to the highly active trans-zeatin by cis–trans
isomerization.
 Based on the half-life of tRNA, it was estimated that tRNA turnover can
account for a significant portion of the total cytokinins in plant tissues .
 critical constraint of the indirect pathway as a major contributor to the
cytokinin pool is the fact that tRNA degradation is not a regulated
process, while the biosynthesis of cytokinins should be precisely
controlled.
 cytokinin synthesis seems to be localized, with centers of high
biosynthetic activity in root tips, shoot meristems and developing seeds,
while tRNA turnover is not tissue-specific.
 So the direct pathway of cytokinin biosynthesis is probably the more
important of the two, but the steps of the pathway remain hypothetical
until genes encoding specific enzymes have been isolated. It is possible
that enzymes and genes somewhat different from those found in the
microbial systems occur in plants.
 The plant and bacterial IPT enzymes differ in the adenosine substrate
used; the plant enzyme appears to utilize both ADP and ATP, and the
bacterial enzyme utilizes AMP.
 The products of these reactions (iPMP, iPDP, or iPTP) are converted to
zeatin by an unidentified hydroxylase. The various phosphorylated forms
can be interconverted.
 Free trans-Zeatin can be formed from the riboside by enzymes of general
purine metabolism. trans-Zeatin can be metabolized in various ways as
shown and these reactions are catalyzed by the indicated enzymes.
METABOLISM OF CYTOKININ-
 Free cytokinins are readily converted to their respective nucleoside and
nucleotide forms. Such interconversions likely involve enzymes common
to purine metabolism.
 Many plant tissues contain the enzyme cytokinin oxidase,which cleaves
the side chain from zeatin (both cis and trans), zeatin riboside, iP, and
their N-glucosides, but not their O-glucoside derivatives.
 Cytokinin oxidase irreversibly inactivates cytokinins, and it could be
important in regulating or limiting cytokinin effects.
 Cytokinin levels can also be regulated by conjugation of the hormone at
various positions.
 The nitrogens at the 3, 7,and 9 positions of the adenine ring of
cytokinins can be conjugated to glucose residues.
 Alanine can also be conjugated to the nitrogen at the 9 positon, forming
lupinic acid.
 The hydroxyl group of the side chain of cytokinins is also the target for
conjugation to glucose residues, or in some cases xylose residues, yielding
O-glucoside and Oxyloside cytokinins.
 O-glucosides are resistant to cytokinin oxidases.
 Enzymes that catalyze the conjugation have substrate specificities for the
sugar donor and the cytokinin bases.
 Only free trans-zeatin and dihydrozeatin bases are efficient substrates;
the corresponding nucleosides are not substrates, nor is cis-zeatin. The
specificity of these enzymes suggests that the conjugation to the side chain
is precisely regulated.
 The conjugations at the side chain can be removed by glucosidase enzymes
to yield free cytokinins, which are the active forms. Thus, cytokinin
glucosides may be a storage form, or metabolically inactive state of these
compounds.
 A gene encoding a glucosidase that can release cytokinins from sugar
conjugates.
 Dormant seeds often have high levels of cytokinin glucosides but very low
levels of hormonally active free cytokinins.
 Levels of free cytokinins increase rapidly, however as germination is
initiated, and this increase in free cytokinins is accompanied by a
corresponding decrease in cytokinin glucosides.
BIOLOGICAL ROLES OF CYTOKININ-
Cytokinins Regulate Cell Division in Shoots and Roots.

 Cytokinins Regulate Specific Components of the Cell Cycle.

 Auxin :cytokinin ratio regulates morphogenesis in cultured tissues.

.Cytokinin modify apical dominance and promote lateral bud growth.

.Cytokinin overproduction has been implicated in genetic tumors.
 Cytokinin delay leaf senescence.
 Cytokinin promote movement of nutrient and chloroplast development.
 Cytokinin promotes cell expansion in leaves and cotyledons.
THANK YOU