1. Plant Tissue Culture?

2. Definition
the culture of plant seeds, organs, tissues, cells, or protoplasts on nutrient media under sterile conditions.
Additional Points: Growth of plant cells outside of an intact plant
A technique essential to many areas of plant science
Culture of individual or groups of cells and whole organs contribute to the understanding of both fundamental and applied science
Cultures can be sustained and maintained as a mass of undifferentiated cells for an extended period of time or regenerated into whole plants

3. How?
Adult plant cells are totipotent, meaning they have the ability to give rise to a fully differentiated plant. Because of this, it is possible to collect cells from a mature plant and use those cells to produce clones of that plant.

4. Advantages Mass production of various plant cultivars
6 million plants per year from one explant.
Much higher production rate than other asexual propagation methods.
Especially beneficial for:
Plants in high demand or valuable plants.
Plants that are slow or difficult to propagate.
Endangered species.

5. Advantages Production of pathogen-free plants
Maintaining disease-free plants by micropropagation.
Continuous year round production
Unaffected by climate

6. Advantages
The original plant is not destroyed in the process - a factor of considerable importance to the owner of a rare or unusual plant.

7. Disadvantages Need for Specialized equipment required
Laminar flow cabinets
Autoclave
Water purification systems
Glassware etc…
Contamination risks
Maintenance of aseptic (sterile) environment difficult.
Rapid spread of contaminants = widespread loss.
Risk of mutation arising
Artificial environment induces mutations.
Responses to tissue culture conditions varies
Trial and error to determine optimum media or conditions。

8. WHAT DO WE NEED? Appropriate tissue A suitable growth medium
Aseptic (sterile) conditions
Growth regulators: especially controlling the ratios of auxins and cytokinins
Frequent subculturing
Emphasize the implications for genetic involvement: Could there be undesirable genes linked to genes influencing tissue culture response?

10. Growth Media Salt Mixtures Macro-nutrient Organic Substances
N, P, K, Ca,Mg,Fe
Micro-nutrient
Zn, Mo, Cu
Organic Substances
Carbon Sources
Sucrose
Vitamins
Adenine
Thiamine
Gelling agents
Agar, agarose
Growth Regulators

11. Preparation 100 ml MS medium
Measure out approximately 70% of the final required volume of tissue culture grade water e.g. 70 ml for a final volume of 100 ml. Select a container twice the size of the final volume.
While stirring the water add 0.44 mg medium and stir until completely dissolved.
Add 3 gm sucrose and stir until completely dissolved
Add desired heat stable supplements (e.g. vitamins, auxins, cytokinins, etc.)
Add additional tissue culture grade water to bring the medium to the final volume.
While stirring, adjust medium to desired pH using HCl or KOH to 5.8.
Add 3 gm agar and heat until the solution is clear.
Dispense the medium into the culture vessels before autoclaving according to your application.
Sterilize the medium in a validated autoclave at 1 kg/cm2 (15 psi), 121 °C, for the time period described under Sterilization of Media Protocol.
Allow medium to cool prior to use.

12. Natural Complexes
Coconut endosperm
Tomato juice
Yeast extracts

13. Commercial Media Formulations
Murashige and Skoog (MS)
White Medium
Nitsch and Nitsch Medium
Woody plant medium