micropropagation in plant biotechnology.pptx

this slide describes about the details of protocol used for micropropagation.

1. M i c r o p r o p a g a t i o n Or Clonal propagation

2. 1 Micropropagation 3 2 Germplasm Preservation 4 Somaclonal variation B r ee d i n g A pp l i c a t i o n s o f T i ss u e Culture 5 5 I n v i t r o H y b r i d i z a t i o n - Protoplast fusion Haploid Production Embryo Culture

3. Micropropagation I n v i t r o C l o n a l P r o p a g a t i o n . M i c r o p r o p a g a t i o n i s t h e p r a c t i c e o f r a p i d l y m u l t i p l y i n g s t o c k p l a n t m a t e r i a l t o p r o d u c e a l a r g e nu m b e r o f p r o g e n y p l a n t s , u s i n g m o d e r n p l a n t t i ss u e c u l t u r e m e t h o d s .

4. CLONE Clone is a plant population derived from a single individual by asexual reproduction. Clonal Propagation is the multiplication of genetically identical individuals by asexual reproduction

5. F e a t u r e s o f M i c r o p r o p a g a t i o n Clonal reproduction Multiplication stage can be recycled many times to produce an unlimited number of clones Easy to manipulate production cycles Disease-free plants can be produced

6. Rapid clonal in vitro propagation of plants: From cells, tissues or organs Cultured aseptically on defined media Contained in culture vessels Maintained under controlled conditions of light and temperatur

7. C o mm e r c i a li z a t i o n o f M i c r o p r o p a g a t i o n 1970 s & 1980 s Murashige (1974) Broad commercial application

8. Starting m a t e r i a l f o r P r o p a g a t i o n

9. S e l e c t i o n o f p l a n t m a t e r i a l 1 Part of plant 2 G e no t y p e 3 P h y s i o l o g i c a l Condition 4 S e a s o n 05 Position on plant 6 Size of Explant

10. 1 Mineral 3 2 Sugar 4 Organic growth factor Medium 5 5 Other Additives Gelling Agent Growth Regulator

11. Physical E n v i r o n m e n t T E M P E R A T U R E MOISTURE LIGHT

12. S T A G E S S e l e c t i o n o f p l a n t m a t e r i a l E s t a b li s h a s e p t i c c u l t u r e Multiplication S h oo t e l o n g a t i o n R oo t i n d u c t i o n / f o r m a t i o n Acclimatization

13. S t e p o f M i c r o p r o p a g a t i o n S t a g e I – E s t a b li s h m e n t S e l e c t i o n o f t h e e x p l a n t p l a n t Sterilization of the plant tissue takes place E s t a b li s h m e n t t o g r o w t h m e d i u m S t a g e I I - P r o li f e r a t i o n T r a n s f e r t o p r o li f e r a t i o n m e d i a S h oo t s c a n b e c o n s t a n t l y d i v i d e d S t a g e II I – R oo t i n g & H a r d e n i n g e x p l a n t t r a n s f e rr e d t o r oo t m e d i a e x p l a n t r e t u r n e d t o s o i l

14. Organogenesis Organogenesis via callus formation Direct adventitious organ formation Embryogenesis Direct embryogenesis Indirect embryogenesis Microcutting Meristem culture (Mericloning) Bud culture M e t h o d s o f M i c r o p r o p a g a t i o n

15. Organogenesis PGRs are prob. the most important factor affecting organogenesis Cytokinins tend to stimulate formation of shoots Auxins tend to stimulate formation of roots The central dogma of organogenesis: a high cytokinin:auxin ratio promotes shoots and inhibits roots high auxin:cytokinin ratio promotes roots and/or callus formation while inhibiting shoot formation

16. Organogenesis The process of initiation and development of a structure that shows natural organ form and function. The ability of non-meristematic plant tissues to form various organs de novo. The production of roots, shoots or leaves. These organs may arise out of pre-existing meristems or out of differentiated cells. This, like embryogenesis, may involve a callus intermediate but often occurs without callus

17. Tissue culture maintains the genetic of the cell or tissue used as an explant. Tissue culture conditions can be modified to cause to somatic cells to reprogram into a bipolar structure. These bipolar structures behave like a true e m b r y o - c a ll e d s o m a t i c e m b r y o s An Embryo is made up of actively growing cells and the term is normally used to describe the early formation of tissue in the first stages of growth. S o m a t i c E m b r y o s

18. The process of initiation and development of embryos or embryo-like structures from somatic cells The production of embryos from somatic or “non-germ” cells. Usually involves a callus intermediate stage which can result in variation among seedlings S o m a t i c E m b r y o g e n e s i s

19. The composition of the culture medium controls the process- auxin (usually 2,4-D) added causes induction, the formation of embrygogenic clumps or proembryogenic masses (PEMs) (induction medium) auxin is deleted and the clumps become mature embryos (maturation medium)

20. e a rl y c e l l d i v i s i o n d o e s n ' t f o ll o w a f i x e d p a tt e r n , u n l i k e w i t h z y g o t i c embryogenesis l a t e r s t a g e s a r e v e r y s i m i l a r t o z y g o t i c e m b r y o s ( d i c o t p a tt e r n ) globular stage (multicellular) heart-shaped stage (bilateral symmetry) –bipolarity torpedo-shaped stage – consists of initial cells for the shoot/root meristem S t a g e s o f d e v e l o p m e n t

21. S t a g e s o f S o m a t i c e m b r y o d e v e l o p m e n t

22. Somatic E m b r y o g e n e s i s Stimulation of callus or suspension cells to undergo a developmental pathway that mimics the development of the zygotic embryo

23. A dv a n t a g e s From one to many propagules rapidly. Multiplication in controlled lab conditions. Continuous propagation year round. Potential for disease-free propagules. Inexpensive per plant once established.

24. Disdvantages Specialized equipment/facilities required. More technical expertise required. Protocols not optimized for all species. Plants produced may not fit industry standards. Relatively expensive to set up.

25. M i c r o p r o p a g a t i o n L i m i t a t i o n s Equipment/facility intensive operation Technical expertise in management positions Protocols not optimized for all species Liners may not fit industry standard Propagules may be too expensive

26. Applications Rapid increase of new varieties. Elimination of diseases. Cloning of plant types not easily propagated by conventional methods. Propagules have enhanced growth features (multibranched character;Ficus, Syngonium)

27. T h a n k Y o u