Plant tissue culture. Micropropagation. Features of micropropagation. Selection of plant material. Physical environment. Steps of micropropagation. Methods of micropropagation. Organogenesis. Somatic embbryogenesis. Stages of somatic embryo development. Micropropagation limitations. Applications. References.

1. Plant Tissue Culture
Micropropagation Stage 2: Shoot Multiplication
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By
KAUSHAL KUMAR SAHU
Assistant Professor (Ad Hoc)
Department of Biotechnology
Govt. Digvijay Autonomous P. G. College
Raj-Nandgaon ( C. G. )

2.  Plant tissue culture.
 Micropropagation.
 Features of micropropagation.
 Selection of plant material.
 Physical environment.
 Steps of micropropagation.
 Methods of micropropagation.
 Organogenesis.
 Somatic embbryogenesis.
 Stages of somatic embryo development.
 Micropropagation limitations.
 Applications.
 References.
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3.  The culture of plant seeds, organs, tissues,
cells, or protoplasts on nutrient media under
sterile conditions.
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4.  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.
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5.  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.
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6.  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
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7. Rapid clonal in vitro propagation of plants:
• From cells, tissues or organs
• Cultured aseptically on defined media
• Maintained under controlled conditions of
light and temperature
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8. Commercialization of Micropropagation 1970s & 1980s
Murashige (1974)
Broad commercial application
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9. Tip
bud
Leaf
Axillary
bud
Internode
Root
Starting material for
micropropagation
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10.  Part of plant
 Genotype
 Physiological condition
 Season
 Position on plant
 Size of explant
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11.  Temperature
 Moisture
 Light
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12.  Stage I –Establishment of Aseptic Culture
◦ Selection of the explant plant
◦ Sterilization of the plant tissue takes place
◦ Establishment to growth medium
 Stage II - Multiplications of shoots
◦ Transfer to proliferation media
◦ Shoots can be constantly divided
 Stage III – Rooting & Hardening
◦ explant transferred to root media
◦ explant returned to soil
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13.  Organogenesis
◦ Organogenesis via callus formation
◦ Direct adventitious organ formation
 Embryogenesis
◦ Direct embryogenesis
◦ Indirect embryogenesis
 Microcutting
◦ Meristem culture (Mericloning)
◦ Bud culture
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14. 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.
The production of roots, shoots or leaves.
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15.  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.
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16.  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
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17.  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
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19. ◦ Technical expertise in management positions
• Protocols not optimized for all species
• Liners may not fit industry standard
• Propagules may be too expensive
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20. • 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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21.  Plant tissue culture- S.S. Bhojwani & Razdan.
 Google-www.google.com.
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