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Cont.

Overall Cell Cycle

A Cell stopped in G1, not necessary to


synthesize DNA & other molecules unless
the cell is determined to enter S-phase. Its
the same with cell in G2, not necessary to
prepare for mitosis unless the cell is going
to divide.
Cells only arrested in G1 & G2 but not in M
and S ( Vant Hoff, 1974).
Cont.
Vant Hoff also showed that cells of Pisum which
were arrested in G1 can be manipulated to stop in
G2.
Substance/chemical originated from the cotyledon
which made the cell stop/arrested in G2. ( G2 Factor-
Trigonelline ( Evans et.al., 1979).
Cells will stop in G2 when this trigonelline is present
or stop in G1 when this substance is absent.
Most legumes, Pisum sativum, Vicia faba, Glycine
max and Phaseolus vulgaris contain high
concentration of this substance ( Trigonelline).
While non-legumes & monocots contain very low
concentration of trigonelline ( Evans & Tramontano,
1984). This indicates that protein synthesis possibly
involves in this mechanism.
Another Cell Cycle Hypothesis
Proposed by Brown ( 1976)- Genes
controlling events in Cell cycle were
arranged in groups. Each group
correspond/related to 4 different phases
(G1,G2, M, S).
Only 1 gene in each group operating at any
one time.
Every group is controlled by gene- Gene
Master
Ref; Cell division in higher Plants
(Yeoman,1976)
Cell Cycle in Vitro
Eriksson ( 1967) was the first person using cell
cycle approach to study plant cells in vitro. He
analysed the cell cycle in cell suspension culture
of Haplopappus gracilis.
Since then the accumulation of cell cycle data
from plant tissues in vitro was very few.
Studies on cell cultured in vitro was to understand
the cell cycle system in intact/in vivo plants.
In few limited studies, comparison was made
between the duration of cell cycles & component
phases in vivo & in vitro.
Cont.
Duration of Cell cyle in vitro usually always longer,
mostly due to G1.
Bayliss (1975) determined the duration of cell
cycle in Daucus carota in the seedlings roots &
also in diploid cell suspension culture
supplemented with 2,4-D. He obtained the
duration of cell cycle in vivo as 7.5 h and 51.2 h for
cells in vitro.
The duration of G1 was 39.6 h in vitro & 1.3 h in
vivo.
Studies on root cells of Zea mays ( Verma, 1980)
showed that the duration of cell cycle was 9.9 h.
Gould (1984) obtained 37 h for cells in vitro.
Cont.

The duration of G1 was 23.9 in vitro


and only 1.7 h in root meristem cells.
These differences portrayed
differences in morphology of the
meristematic cells in vivo as
compared to cells in vitro.
The duration of cell cycle in vivo & in
vitro can be summarised as follows;
Table 1

Species Duration G1 (h) S (h) G2 (h) M (h) Reference


Dc/T (h)
Daucus 7.5 1.3 2.7 2.9 0.6 Bayliss,
carota 1975
(root
cells)
Cultured 51.2 39.6 3.0 6.2 2.4
cells

Haplopap 10.5 3.5 4.0 1.4 1.6 Sparvoli,


pus Gray&
gracilis Kaufman,
(Root 1966
cells)
Cultured 22 9.3 6.4 4.9 1.4 Ericksson,
cells 1967
Zea mays 9.9 1.7 5.0 2.1 1.1 Verma,
Root cells 1980

Cultured 37.0 23.9 7.1 3.9 2.1 Gould,1984


cells
Cell Cycle Regulation in Plants
Cell division plays a crucial role during all phases
of plant development.
The molecular analysis of cell division & its
regulation in plants lags far behind yeast &
animals.
Genes such as ras and p53 are players in yeast &
animals that activate the cell-cycle engine.
In all eukaryotes, the biochemical machine that
controls progress through the cell cycle consists of
a catalytic protein kinase & an activating cyclin
subunit.
This complex but neither protein alone, has protein
kinase activity.
Stepwise changes specifically regulate
progression through the cycle.
Cont.
The proteins encoded by S.cerevisiae
CDC28 and the S. pombe Cdc2 genes
are the prototypic CDKs required at
both the G1/S and G2/M transitions.
In animals CDK1(=cdc2) is essential for
the G2/M transition, whereas CDK2 &
CDK3 are required for the G1/S
transition.
These 3 CDKs share a short amino acid
sequence PSTAIRE
Cont.
Several related kinases with homology but
incomplete sequence identity to the
PSTAIRE domain have been identified.
Cyclins were the first identified in marine
invertebrates as proteins, when injected
into frog oocytes could induce meiosis.
The observation that S. pombe cdc13
gene, which encodes a cyclin, genetically
interacts with cdc2 established a tight
functional link between the components of
the cell cycle engine. Since then, an
increasing no. of cyclins have been
identified in eukaryotes.
Cont.
In S. cerevisiae 3 cyclins ( CLN1, CLN2,CLN3) are
expressed & presumably function in G1, 2 are
expressed in S phase
( CLB5, CLB4) & 4 are expressed in mitosis
( CLB1, CLB2, CLB3, CLB4).
In animals, the C-D- and E- class cyclins are
expressed in G1, cyclin A is expressed in the S
&G2, B cyclins are expressed in the G2 & early M
phases.
In most cells the decision whether to commit to
division is made during G1, by G1 cyclins.
Cont.

Cyclins are involved in


determining the substrate
specificity of CDKs as well as in
targeting CDK activity to specific
subcellular compartments during
the cell cycle.
Cell Division in Plants
Plant genes homologous to components of the cell
cycle engine have been cloned recently but the
function of these putative cell cycle regulators has
not yet been examined in plants. Therefore, at
present the functions of these genes are unknown.
CDKs have been cloned from pea, alfalfa, maize,
Arabidopsis.
Plant cyclins have been cloned from carrots,
soybean, alfalfa & Arabidopsis.
The soybean & Arabidopsis gene products
promote nuclear envelope breakdown when the
corresponding mRNAs are injected into Xenopus
oocyctes, indicating their function as mitotic
cyclins.
Cont.
All plant cyclins share homology to both A-
& B- type cyclins. It is unclear whether
these plant cyclins play roles during S
phase as reported for animal A type
cyclins, as well as during the G2 & M
phases as shown for B-type cyclins. A-type
cyclins have been found only in animals.
Therefore, plant AB- cyclins may be early
members of an evolutionarily more
divergent cyclin gene family. No G1 cyclin
has yet been reported from plants.
Cont.

The expression of cdc2 homologs


is higher in mitotically active
tissues such as floral buds and
low or undetectable in mature,
nonproliferating organs such as
leaves.

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