OSMOTIC EFFECT OF

MEDIA INGREDIENTS
Group 2: Chu Hoang Trung Tin
Nguyn L Thuy Truc
Thi Kim Tin
Hang Anh Tai

CONTENT
I. OSMOTIC AND WATER
POTENTIALS
II. THE OSMOTIC POTENTIAL OF
TISSUE CULTURE MEDIA
III.EFFECTS AND USES OF
OSMOLYTES IN TISSUE CULTURE
MEDIUM

I. OSMOTIC AND WATER

POTENTIALS
: water potential Pure water: =0
= s + p
s : solutions potentials
p : pressure potential
water always flow from
high to low water potential

OSMOTIC AND WATER

POTENTIALS
Osmolality: The unit of the
osmolality of a solution exerting
an osmotic pressure equal to that
of an ideal nondissociating
substance which has a
concentration of one mole of
solute per kilogram of solvent.
s = -I C RT
The addition of solutes to a
solvent has been said to
reduce the osmotic potential
of solutions.

OSMOTIC AND WATER

POTENTIALS
cell = s + p + m
s : solutions potentials
p : pressure potential
x
m : cell wall matrix
potential
= cell
outside
= cellA cellB

II. THE OSMOTIC POTENTIAL

OF
TISSUE CULTURE MEDIA
The total osmotic potential of
solutes

S= S macronutrients + S sugars
Sugars are responsible for much
of the osmotic potential of
normal plant culture media

THE CONTRIBUTION OF THE

GELLING AGENT

The water potential of media

solidified with gels is more
negative than that of a liquid
medium, due to their matric
potential

THE CONTRIBUTION OF
NUTRIENTS SALTS
0.1M KCL dissolved in water =>
0.1 mole K+ and 0.1 mole Cl 0.2 theoretical osmolality
Several different compounds are
dissolved together =>
Dissociation may not be
complete
The macronutrients contribute
most to the final osmotic

THE CONTRIBUTION OF
SUGARS

THE CONTRIBUTION OF
SUGARS

DECREASING OSMOTIC
POTENTIAL WITH OTHER
OSMOTICA

Sugars can act as osmoregulants.

Inhibits water uptake.
Employed for the deliberate
modification of osmotic potential.
Cannot be assimilated.
Lacking other biological effects.
Ex: Mannitol, sorbitol,
polyetheleneglycol, sodium sulphate,
sodium chloride, etc.

III.EFFECTS AND USES OF

OSMOLYTES IN TISSUE
CULTURE MEDIUM

Osmolytes
are compounds
affectingosmosis.
play a role in maintaining
cell volume andfluid
balance.

1. Protoplast isolation and

culture:
A
protoplast
is
aplant,bacterialorfungalcell that
had
itscell
wallcompletely
or
partially
removed
using
either
mechanical or enzymatic means.
In plant cells:
The
osmotic
potential
is
counterbalanced by the pressure
potential exerted by the cell wall.

(b) Loss of water makes

the cell limp and shrinks
the cell membrane away
from the cell wall
Figure. Osmosis in(plasmolyse
plant cells
cells).

(a) Water entering

the cell by osmosis
inflates the cell and
makes it rigid.

 To safely remove the cell wall

without damaging the plasma
membrane: placing the cells in a
solution of lower water potential
than that of the cells.
Glucose, sucrose and especially
mannitol and sorbitol, are usually
added to media.

2. Osmotic effects on
growth:
Solutions of different concentrations
exert their effect on growth by their
nutritional value, and through their
varying osmotic potential.
Decreasing the osmotic potential
(usually by adding mannitol) +
lowering of the temperature =>
reduce
the
growth
rate
for
preservation of valuable genotypes

3. Osmotic effects on
morphogenesis

[SUGA
R]

Correct water
potential
Apogamous buds on
ferns
Root formation and
root growth

Somatic
embryogenesis
Storage organ
formation
Anther culture

Correct water potential

Tobaco callus growth and shoot

regeneration : optimum at [sucrose]
= 3%
Wheat
callus
growth
and
adventitious shoot formation in MS
medium with 2% sucrose.
Very small potato calluses formed
can survive in 1 or 2% sucrose ><
full green only on 0.2- 0.5%.

Apogamous buds on
ferns

The stimulatory effect of

glucose action as a
respiratory substrate.
Inhibition by an
excessively depressed
osmotic potential.

Root formation and root

growth
An
excessive
sugar
concentration can inhibit root
formation
Although 4% (and occasionally
8%) sucrose used for isolated
root culture, 2% used in the
great majority of cases

Somatic embryogenesis
High sucrose levels in media
for the culture of zygotic
embryos if they are isolated
when immature.
Embryos which are more fully
developed when excised,
grow satisfactorily in a
medium with 10-30 g/l sugar.

Storage organ formation &

Anther culture
At high concentration, sucrose :
Formation of tubers, bulbs and corms
The addition of 5-20% sucrose: assist
the development of somatic embryos
from pollen microspores

High concentration of mannitol:

pretreatment before the culture of
barley anthers ,pollen tobacco
anthers and pollen.

[ PHYTOHORMONE]
Auxin : very important influence on
the growth and morphogenesis.
ABA + an osmoticum prevents
precocious germination in white
spruce and allows embryo
development.
Storage organ formation might be
mediated by ABA since osmotic
stress induces ABA => bulb and

[ION]
Callus cultured in the presence of
sodium sulphate retained its shootproducing capacity over a long
period.
Media of small osmotic potential:
employed for the induction and
growth of roots on micropropagated
shoots
Root initiation: