PLANT TISSUE CULTURE 1

Totipotency

Differentiation

Dedifferentiation

Redifferentiation

What are the Basic concepts of plant tissue culture?

Totipotency

The property of live plant cells that they have the GENETIC POTENTIAL when cultured in nutrient medium to GIVE RISE to a COMPLETE INDIVIDUAL PLANT.

Differentiation

The process of biochemical and structural changes by which CELLS BECOME SPECIALIZED in FORM and FUNCTION.

• The FURTHER DIFFERENTIATION of already differentiated cell INTO ANOTHER TYPE of CELL.

• For example, when the component CELLS of CALLUS have the ability to FORM a WHOLE PLANT in a NUTRIENT MEDIUM, the phenomenon is called redifferentiation.

Dedifferentiation

• The phenomenon of the REVERSION of MATURE CELLS to the MERISTEMATIC state leading to the FORMATION of CALLUS is called dedifferentiation.

• Process associated with RE-ENTRY into CELL CYCLE trans/redifferentiation or even cell death.

G. Haberlandt

In 1898, German Botanist ______ successfully CULTURED INDIVIDUAL PLANT CELLS, isolated from different tissues.

Gauthret

White

Nobecourt

• But only during 1934 to 1939, a foundation of plant tissue culture was laid down by three scientists: ____, ____, and ____ due to discovery of plant growth regulators such as AUXINS and VITAMINS.

1960

• Year when IN-VITRO culture of PLANT CELLS, TISSUE and ORGANS was reasonably WELL DEVELOPED.

• Consequently, MEDIA and CULTURE TECHNIQUES for variety of plant materials become KNOWN.

Plant Tissue Culture

It is used to DESCRIBE the IN-VITRO and ASEPTIC GROWTH of any PLANT PART on a TISSUE CULTURE MEDIUM.

Culture vessels

• PLANT TISSUE STUDIES; Erlenmeyer flask, petri dish, and culture tubes.

• provides a BARRIER against contamination to PROTECT the CULTURES from the external environment while maintain an adequate internal environment.

Culture medium

• important media used for all purpose experiment are MURASHIGE and SKOOG medium (MS medium), GAMBORG medium (B5 medium), WHITE medium (W medium) and NITSCH medium.

• the culture medium is closed with cotton plug or aluminum foil sheet; pH medium is adjusted to 5.8.

Sterilization

• technique employed to GET RID of the MICROBES in the culture media and plant tissue.

• culture medium should be keep in autoclave 121 DEGREE CELSIUS, 15MINS; plant tissue is to be surface sterilized.

Inoculation

• TRANSFER of PLANT (root, stem or leaf); aseptic condition under laminar air flow chamber.

• flamed and cooled forceps are used for transfer of plant materials.

Incubation

• the inoculum is incubated at 26 degree Celsius with light intensity at 2000 to 4000 lux.

• 16hrs of light and 8hrs of darkness

• the EXPLANT is INDUCED to FORM CALLUS

• CALLUS formation is induced through AUXIN, CELL ELONGATES and CYTOKININ induces CELL DIVISION = masses of cells are formed.

Morphogenesis

• FORMATION of NEW ORGANS FROM the CALLUS under the influence of auxin & cytokinin.

• young plantlet

somatic embryo

• ROOTS and SHOOTS are DIFFERENTIATED FROM the CALLUS → such embryo is called?

• FORMATION of NEW ORGANS such as SHOOT and ROOT.

• DEVELOPMENT of SHOOT from callus ~ CAULOGENESIS; FORMATION of ROOT ~RHIZOGENESIS

Caulogenesis

• DEVELOPMENT of SHOOT from CALLUS

Rhizogenesis

• FORMATION of ROOT from CALLUS

• FORMATION of EMBRYO.

• Embryo arise from somatic callus = somatic

  embryos or embryoids or somaclonal embryos.

somatic embryos or embryoids or somaclonal embryos

• EMBRYO arise from SOMATIC CALLUS

Hardening

• EXPOSING the PLANTLET to the NATURAL ENVIRONMENT

• TRANSFERRING of PLANTLET to the SOIL.

5.8

Culture medium is adjusted to ph of?

121

culture medium should be keep in autoclave ___ degree Celsius in 15 MINS

26

• the inoculum is incubated at ____ degree Celsius with light intensity at 2000 to 4000 lux.

16

8

• ______hrs of light and _____hrs of darkness

auxin

cell elongates

cytokinin

induces cell division