Plant tissue culture involves growing plant cells, tissues or organs in sterile conditions on a nutrient culture medium. It has four key stages: establishment, multiplication, rooting, and acclimatization. The establishment stage involves culturing an explant on a nutrient medium to encourage cell division. In multiplication, the explant proliferates to form a callus or multiple shoots. Rooting occurs when shoots are transferred to a medium promoting root growth. Finally, in acclimatization, rooted plantlets are hardened off before being planted.

1. PLANT CELLAND
TISSUE CULTURE
Presented by: Talha Saleem

2. The growth and development of plant seeds, organs,
explants, tissues, cells or protoplasts on nutrient media
under sterile conditions.

3. • 1. Environmental condition optimized (nutrition, light, temperature).
• 2. Ability to give rise to callus, embryos,
adventitious roots and shoots.
• 3. Ability to grow as single cells (protoplasts, microspores,
suspension cultures).
• 4. Plant cells are totipotent, able to regenerate a whole.
Characteristic of Plant Tissue Culture
Techniques

4. Tissue culture has several critical requirements:
• Appropriate tissue (some tissues culture better than
others)
• A suitable growth medium containing energy sources
and inorganic salts to supply cell growth needs. This can
be liquid or semisolid
• Aseptic (sterile) conditions, as microorganisms grow
much more quickly than plant and animal tissue and can over run a
culture

5. Media
• Must be sterile
• Autoclave at 250 F at 15 psi for 15 minutes
• pH of media: usually 5.0-5.7
• Frequent sub culturing to ensure adequate nutrition and to avoid the build up of
waste metabolites.

6. Explant
• This means to simply cut-out a very small piece of leaf or stem tissue, or even isolate
individual cells, and them in a tissue culture container.
• The tissue has to be surface-sterilized so it will not have any contaminating bacteria or
fungus.
• It is then placed inside the tissue culture vessel (dish, jar, etc.) containing a gel called agar.
• In the agar dissolved all the sugar, nutrients and plant growth regulators the explant
needs

7. Explant
• Species dependent
• Physiological age - young portions of plant are
most successful.
• Important features : size & source - some tissues
are better than others

8. Plant Tissue Culture
Plant
Tissue
Culture
Your Text Here
Rooting
(stage III)
Acclimatization
(stage IV)
Your Text Here
Multiplication
(stage II)
Establishment
(stage I)

9. Growth stages of micro propagation
• Establishment (stage I)
• choice of explant
• disinfestation and plating
• Multiplication (stage II)
• axillary branching and shoot proliferation
• cytokines included to stimulate shoots
• Rooting (stage III)
• auxin included to stimulate roots
• Acclimatization (stage IV)

10. Rinse with Sterile Water Three
Times for Five Minutes Each

11. Sterilize Forceps and Scalpels
Flame and Cool Burn-Off Alcohol

12. Preparing the Tissue for
Culture
Cut the Ends Trim the Petiole

13. Stage I: Establishment
• Establishment of the explant in a culture medium. The medium sustains the
plant cells and encourages cell division. It can be solid or liquid
• Each plant species (and sometimes the variety within a species) has particular
medium requirements that must be established by trial and error

14. • Multiplication- The explant gives rise to a callus (a mass of loosely
arranged cells) which is manipulated by varying sugar
concentrations and the auxin (low): cytokinin (high) ratios to
form multiple shoots.
• The callus may be subdivided a number of times.
Stage II: Multiplication

15. Dividing stage II cultures

16. Stage III: Rooting
•Root formation - The shoots are transferred to a growth
medium with relatively higher auxin: cytokinin ratios

17. Stage IV: Acclimatization of
Plantlets
• The rooted shoots are potted up (deflasked) and ‘hardened off’ by
gradually decreasing the humidity
• This is necessary as many young tissue culture plants have no
waxy cuticle to prevent water loss

18. REFERENCES
• https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-
biology/plant-tissue-
culture#:~:text=Plant%20tissue%20culture%20is%20defined,light%2C%20tempe
rature%2C%20and%20humidity.
• http://faperta.ugm.ac.id/newbie/download/pak_tar/Plantbiotechnology/biotek-
pemuliaan/plntbiotech20101.ppt