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Micropropagation

Micropropagation is the process of rapidly multiplying plant materials using modern tissue culture methods under sterile conditions. The ratio of two key plant hormones, auxin and cytokinin, determines whether roots or shoots will develop. Micropropagation allows for large-scale asexual cloning of genetically identical plant materials and is used for commercial purposes such as preserving plant germplasm and producing disease-free plants.

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Brigitte Reyes
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311 views

Micropropagation

Micropropagation is the process of rapidly multiplying plant materials using modern tissue culture methods under sterile conditions. The ratio of two key plant hormones, auxin and cytokinin, determines whether roots or shoots will develop. Micropropagation allows for large-scale asexual cloning of genetically identical plant materials and is used for commercial purposes such as preserving plant germplasm and producing disease-free plants.

Uploaded by

Brigitte Reyes
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPT, PDF, TXT or read online on Scribd
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 The culture of plant seeds, organs, tissues,

cells, or protoplasts on nutrient media under


sterile conditions.
 Two Hormones Affect Plant Differentiation:
◦ Auxin: Stimulates Root Development
◦ Cytokinin: Stimulates Shoot Development

 Generally, the ratio of these two hormones can


determine plant development:
◦  Auxin ↓Cytokinin = Root Development
◦  Cytokinin ↓Auxin = Shoot Development
◦ Auxin = Cytokinin = Callus Development
 Micropropagation
 Germplasm Preservation
 Somaclonal variation
 Embryo Culture
 Haploid Production
 In vitro Hybridization- Protoplast fusion
 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.
 Clone is a plant population derived from a
single individual by asexual reproduction.

 ClonalPropagation is the multiplication of


genetically identical individuals by asexual
reproduction.
 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


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 temperature
Commercialization of Micropropagation 1970s & 1980s
Murashige (1974)
Broad commercial application
Tip
bud
Leaf

Axillary
bud

Internode

Root

Starting material for


micropropagation
 Part of plant
 Genotype
 Physiological condition
 Season
 Position on plant
 Size of explant
 Minerals
 Sugar
 Organic ‘growth factors’
 Growth regulators
 Gelling agent
 Other additives
 Temperature
 Moisture
 Light
1. Selection of plant material
2. Establish aseptic culture
3. Multiplication
4. Shoot elongation
5. Root induction / formation
6. Acclimatization
 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
 Organogenesis

◦ Organogenesis via callus formation


◦ Direct adventitious organ formation
 Embryogenesis

◦ Direct embryogenesis
◦ Indirect embryogenesis
 Microcutting

◦ Meristem culture (Mericloning)


◦ Bud culture
 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
◦ a high auxin:cytokinin ratio promotes roots and/or
callus formation while inhibiting shoot formation
 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.
 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.
 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
 Somatic embryogenesis is a useful regeneration
pathway for many monocots and dicots, but is
especially useful for the grasses
 Types of embryogenesis

◦ zygotic embryogenesis – the result of normal


pollination and fertilization
◦ somatic embryogenesis – embryos from
(cultured) sporophytic cells , that is embryos
arise indirectly
 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)
 Stages of development
◦ early cell division doesn't follow a fixed
pattern, unlike with zygotic embryogenesis
◦ later stages are very similar to zygotic embryos
(dicot pattern)
 globular stage (multicellular)
 heart-shaped stage (bilateral symmetry) –
bipolarity
 torpedo-shaped stage – consists of initial cells
for the shoot/root meristem
Somatic Embryogenesis
Stimulation of callus or suspension cells to undergo a
developmental pathway that mimics the development of
the zygotic embryo.
• 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.
• 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.
• 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
• Rapid increase of stock 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)

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