DYA40133

CHAPTER 4
HARDENING AND
ACCLIMATIZATION

KHAIRUNISA BINTI AB AZIZ

POLITEKNIK JELI, KELANTAN
TABLE OF
CONTENT Performed the process of hardening and
acclimatization of micro propagated plants to
ex vitro condition.
COURSE LEARNING OUTCOMES

Students will be able to :

4.1.1 Performed the primary and

secondary hardening sterilization
instrument
 Types Of Hardening
Secondary
• Carried out in shade house
• Plants are transferred to polybags filled with potting
mixture and grown under shaded house for 6 – 8
weeks
• Factors affecting secondary hardening:
Primary • Utilization of polybags for raising the plants
• Size of polybags
• Carried out in laboratory or green house
• Medium used for producing sturdy plants
• Increase survival rates of cultured plants
• Application of nitrogen fixator and solubilizer
• Achieve optimal:
• Amount of plants per area of shaded house
• root system .
• height and girth
• leaves and chlorophyll content
 Step In Primary
Hardening

Remove and Grow the

washing away Introduce Cover the
plantlets in the
the medium (to plantlets into plantlets with
reduce the laboratory or
jiffy-7 plastic bag
contamination ) greenhouse
 Step In Secondary
Hardening

Moisten the
Push the
Move medium in
roots parts of Watering
plantlets from polybags and Placed the
the plantlets immediately
in vitro and make a hole plantlets into
into the hole after
carry to the at the center the polybag
and fill up the transplanting
nursery of polybag (2
hole
inches deep)
Instruments Used In
Hardening
COURSE LEARNING OUTCOMES

Students will be able to :

4.1.2 Acclimatization strategies for

micro propagated plants
 Definition of
Acclimatization

A process of adaptationof an
organism to an
environmental change
 “Process of gradually accustoming plants to
changing environmental conditions of relative
humidity, temperature, light, moisture and medium
Concept of nutrition besides changing levels of contamination
acclimatization during a series of the different stages along the
course of culturing to weaning”
 Need of Acclimatization

Necessary to aid plantlets

to initiate development of
cuticles, stomata/root
Nursing of the plantlets functions before
for final transfer to the transferring to the field
growth complex
field beds High humidity inside the environment
culture vessels will keeps
the cuticles less
developed; stomata does
not operate properly,
sometimes misinformed or
totally absent
 HOW TO EXPOSE PLANTLETS TO
EX VITRO CONDITIONS

• Media:
• Peat-based medium are preferred
• 70 – 80 % quality milled peat
vermiculite or bark
• Avoid coarse and large particles when using bark
• pH adjusted to 5.8 – 6.1 by using dolomitic limestone
• Use low concentration of slow
release fertilizer trace elements)
• Pre-moisturize media before filling into trays
• Light and temperature:
• Keep in mind, plantlets in laboratory
only grown under 2.2 –
6.5 kLux with uniform temperature
(25 °C)
• Must protect plantlets from brighter
greenhouse environment and
extreme temperatures
• Initial environment should be kept at
26 – 30 °C (day) and 21
– 24 °C (night) where bottom heat will
help induce rooting
• Contamination and planting aids:
• Media used for culturing must be
washed gently or shaken off to prevent
the growth of unwanted microorganism
(contains high sugar)
• Use dibble when planting (forceps for
small cuttings)
• Place plantlets just deep enough to
anchor them into the medium
• Use a spray bottle of water to mist the
plantlets during handling
• Water plantlets thoroughly but gently
• Regular fungiside should prevent
losses from root and
stem rot
diseases
• Humidity and fogging system:
• Plantlets from the laboratory requires high humidity
to survive
(no cuticle)
• Mist or fogging system can be used to prevent
desiccation (gradually increase the mist
interval)
• Cover trays with clear lids, layer of
polyethylene film or cheesecloth
• Provide ventilation to prevent heat build up
• Provide additional shading (increase light level until
plantlets
reach its normal light requirements)
• Do not allow medium to be waterlogged
Thank You