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Presented by
ASHAMOL T.C
Plants have the ability to reproduce
asexually
This natural ability is the basis of
micropropagation
Totipotency-Ability of living cell of a
multicellular organism to develop
independantly in to an organism when
supplied by suitable nutrient medium
Which parts of the plant can be used for
micropropagation?
Meristems, shoot and root tips, leaf tissue
anthers, embryos, flowers, virtually all parts of a
plant can be used.
1902-Haberlandt -cultured Isolated single palisade cell
1904-Hanning Robins -Embryo culture 1922- Hanning Robins-Root tip culture 1934-Gautherets-cambial tissue culture 1935-Snow-plant growth hormones
isolated 1962- Murashige and Skoog –formulation
of a well defined medium
WASHING AND CLEANING AREA
MEDIA PREPARATION ROOM
TRANSFER AREA
GROWTH AREA/INCUBATION ROOM
Water tap and sink
Distillation unit
Autoclave
Hot air oven
Gas stove
Chemicals in alphabetical order
Refrigerator
pH meter
Electronic balance/weighing balance
Glass pipettes/measuring cylinder
Glassware
Magnetic stirrer
Laminar Air Flow
Spirit Lamp
Forceps
Sterile Blade
Lighter
Rectified Spirit
FORCEPS
Culture racks
Light…. Fluorescent tube light
Temperature
pH
PHYSICAL
LIGHT
TEMPERATURE 26+2 / 26-2
HUMIDITY
CHEMICAL
INORGANIC
• Macro nutrients -> N, P,K,Ca,S,Mg
• Micro nutrients –> B,Co,Cu ,Mn, Mo, I,Zn
ORGANIC
Sugars, Amino acids, Vitamins, Growth hormones
Stock solution preparation
Add Stock and chemicals to
distilled water as per formulation
Make up to desired concentration
Add hormones
Adjust pH upt o 5.8
Add agar 0.6-0.7% (6-7g/l) Melting of Agar Pouring of medium to culture vessels Autoclaving of medium
AUTOCLAVING
5kg/cm2 Pressure Temperature 1210 C
18-20 Min
1. Medium
2. Sterile petridish
3. Sterile blade
4. Sterile distilled water
5. Spirit lamp
EXPLANTATION
CLEANING AND WASHING OF EXPLANTS
SURFACE STERILISATION
WASHING IN STERILE DISTILLED WATER FOR THREE TO FOUR TIMES
PROCEDURE OF PLANT TISSUE
CULTURE
CUT BOTH ENDS OF PLANT
INOCULATION TO NUTRIENT
MEDIA
PLUGGING
INCUBATION
Stage one – Explant establishment or initiation
Stage two – Multiplication
Stage three – Rooting
Stage four – Acclimatization or hardening off
Selection of explants
Surface sterilization
Incubation of cultures
Rapid multiplication of tissues
Newly formed buds dissected out
Transferred to subculture medium
PRE TREATMENT FOR TRANSFER TO SOIL
Growth regulators for root initiation added
Light intensity is lowered
Later stage intensity increased upto 1o,ooo lux to promote autotrophic growth
Hardening of plant by
keeping under high
humidity and shade
SHOOT BUD
ELONGATION
SHOOT BUD AFTER TWO WEEKS
SHOOT PROLIFERATION
ROOT PROPAGATION
PLANTLETS
Expensive
Particular skills are needed
Contamination due to incorrect sterilization
Medium is specific for many species
Introduction of unknown pathogens in the medium
To regenerate plants from single cells or plant tissues
To produce large quantities of identical plants
To save species from extinction
To produce plants with enhanced stress resistance
To produce new plant varieties
To make money
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