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