brief introduction to cell theory plant tissue culture techniques,concept of totipotecny,callu culture and sterilizatio techniques and media component

1. PLANT BIOTECNOLOGY

2. TOPICS TO BE COVERED Cell Theory Plant Tissue Culture Techniques Callus Tissue & Organogenesis Principles Of Growth Plant Regeneration Concept Of Totipotency Of Cells Plant Tissue Culture Lab Culture Media Media Components Aseptic Techniques Sterilization Techniques Points To Remember

3. Cell Theory Cell theory states that: All living things or organisms are made of cells & their products. New cells are created by old cells dividing into two. Cells are basic building units of life.

4. Plant tissue culture techniques

5. Callus tissue & organogenesis Callus Formation Is Found On Angiosperms, Gymnosperms, pteridophytes& Bryophytes Callus Contains No Organized Meristem Callus Is Somewhat Abnormal Tissue which Has Potentiality To Produce Normal Roots & EmbroidsIn Turn It Develops Into Plantlets. Callus May Be Hard due To Lignifications Of Cell Walls Or Brittle And Sometimes Soft. Callus on a wounded plant parts or on a culture medium is made up of an amorphous , aggregate of loose parenchyma cells which proliferate from mother cells.

6. Organogenesis is the development adventitious organs or primordia (embroid) from undifferentiated cell mass (callus)in tissue culture.

7. Principles of growth Growth Self –multiplication of living materials, the protoplasm itself. Increase in size (volume/length) due to cell division and subsequent enlargement. Increase in dry weight. Development Defined as an ordered change or progress, often towards a higher, more ordered or more complex state.

8. why growth occurs? Expressed as division of a cell to form two cells and the enlargement of newly divided cells.

9. Growth kinetics First rapid phase. Maximum growth rate phase. Last phase.

10. Power of regeneration of plants

11. Concept of totipotency As cell divide mitotically, they do so eqautionally to produce daughters cells. G.Haberlandt’s claimed that one day it could be possible to rear plants from isolated would be rarely surviving cells of flowering plants. He also sated that out of surviving somatic cells artificial embryos would be reared asexually Therefore every cell within the plant has a potential to regenerate into a whole plant.

12. Plant tissue culture lab Media preparation room Culture media, washing powder/liquid disinfectants. Aseptic transfer chamber area Environmentally controlled culture room Analytical room Acclimatization room Miscellaneous items ( air conditioner, marker, match box, burner, etc.)

13. Lab instruments pH meter Balances Electronic hot air over Microscopes Centrifuge Filter sterilizing equipment Laminar air flow (LAF) cabinet.

14. CULTURE MEDIA A Nutrient media generally contains inorganic salts, vitamins, growth regulators, a carbon source & gelling agent. Others include –organic nitrogen, hexitols, amino acids, antibiotics & plant extracts. Nutrition of callus: Chemical factors: minimal & plant growth regulators Environmental factors: light temperature, humidity & genetic constitution or genotype of the plant.

15. Media components

18. B) ORGANIC COMPOUNDS

19. C) GROWTH REGULATORS

23. Aseptic techniques Sterilization : destruction of living matter Disinfectant : chemical agent used to kill pathogens without sterilizing matter to which chemical is applied Sanitation: substantially reducing & then maintaining the micro-organism population in air & on objects in lab to acceptable levels.

24. Sterilization of plant tissues Sodium hypochloride (NaOCl) : 0.025%-0.25% Calcium hypochloride (CaOCl): Hydrogen peroxide (H2O2): 3% -10 % Bromium water: 1% - 2% Silver nitrate (AgNO3) : 1% Mercuric chloride (MgCl2): 0.1 % - 1.1 %

26. Points to remember Cell theory aspects describing the fundamentals of the “CELL” CALLUS tissue Totipotency Growth & development Media components Sterilization technique

27. Thank you