Plant Cell and Tissue Culture

This Lecture is presented by our volunteer Talha Saleem, he is from Karachi Pakistan, and he is covering Plant Cell and Tissue culture topic.
For Video: https://www.youtube.com/watch?v=xoD2cZsTl9g

1. Plant cell and tissue culture Presented by: Talha Saleem

2. Plant Tissue Culture 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%20temperature%2C%20and%20humidity . http:// faperta.ugm.ac.id/newbie/download/pak_tar/Plantbiotechnology/biotek-pemuliaan/plntbiotech20101.ppt