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

1. Micropropagation
Or
Clonal propagation

2. 01 Micropropagation
03
02 Germplasm Preservation
04
Somaclonal variation
BREEDING APPLICATIONS OF TISSUE CULTURE
05
05
In vitro Hybridization-
Protoplast fusion
Haploid Production
Embryo Culture

3. MICROPROPAGATIO
N
In vitro Clonal Propagation.
Micropropagation is the practice of
rapidly multiplying stock plant material to
produce a large number of progeny
plants, using modern plant tissue culture
methods.

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. FEA TURES OF M IC ROP ROPA G A TION
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. Commercialization of Micropropagation 1970s & 1980s
Murashige (1974)
Broad commercial application

8. STARTING
M ATERIA L
FOR
PROPA G ATIO
N

9. SELECTION OF PLANT
MATERIAL
01 Part of plant
02 Genotype
03 Physiological
Condition
04 Season
05 Position on plant
06 Size of Explant

10. 01 Mineral
03
02 Sugar
04
Organic growth factor
MEDIUM
05
05
Other Additives
Gelling Agent
Growth Regulator

11. Physical
Environment
TEMPERATUR
E MOISTURE
LIGHT

12. STAGES
1.Selection of plant material
2. Establish aseptic culture
3.Multiplication
4. Shoot elongation
5. Root induction /formation
6. Acclimatization

13. STEP OF
MICROPROPAGATION
Stage I–Establishment
Selection of the explant plant
Sterilization of the plant tissue takes place
Establishment to growth medium
Stage II - Proliferation
Transfer to proliferation media
Shoots can be constantly divided
Stage III– Rooting & Hardening
explant transferred to root media
explant returned to soil

14. Organogenesis
1.Organogenesis via callus formation
2.Direct adventitious organ formation
Embryogenesis
1.Direct embryogenesis
2.Indirect embryogenesis
Microcutting
1.Meristem culture (Mericloning)
2.Bud culture
M ETHODS OF
M ICROPROPAG ATION

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

16. ORGANOGENESI
SThe 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
embryo - called somatic embryos
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.
SOM ATIC
EM BRYOS

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
SOM ATIC
EM BRYOG ENESIS

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. early cell division doesn't follow a
fixed pattern,unlike with zyg otic
embryogenesis
later stag es are very sim ilar to
zyg otic em bryos (dicot pattern)
globular stage (multicellular) heart-shaped stage (bilateral symmetry)
–bipolarity torpedo-shaped stage – consists of initial cells for the
shoot/root meristem
STAG ES OF
DEVELOPM ENT

21. STAG ES OFSOM ATIC EM BRYO
DEVELOPM ENT

22. SOMATIC
EMBRYOGENESI
S
Stimulation of callus or
suspension cells to
undergo a
developmental pathway
that mimics the
development of the
zygotic embryo

23. ADVANTAG 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. DISDVANTAGE
S
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 ICROPROPAG ATION
LIM ITATIONS
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. APPLICATION
S
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. THA NK YOU