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and Micropropagation
Lecture Outline-Introduction to Tissue Culture
z In developing countries:
– Rapid growth industry
– Production of food, fiber, forestry and medicine
– Future- ornamental exports?
Micropropagation vs. Plant Tissue Culture
z Micropropagation-
–– The production of whole plants from small sections of a plant,
called an “explant”.
“explant”.
z
z Stem
Stem tip
tip
z
z Node
Node
z
z Meristem
Meristem
z
z Embryo
Embryo
z
z Seed.
Seed.
z Plant tissue culture-
–– Inclusive term for growth of cells and tissues in a sterile
microenvironment
z
z undifferentiated
undifferentiated plant
plant cells
cells
z
z plant
plant callus
callus
z
z plant
plant tissue
tissue
Micropropagation
Explant
Any part of the plant (cells, tissue,
organs etc.) used to start in vitro cultures.
Micropropagation
Advantages:
z Mass production of various plant cultivars
– Widespread availability and low cost of exotics
– 10 66 plants / year from a single explant
– Compare with scale of other asexual techniques
–– Example:
Example: Hosta
Hosta division
division
– Especially beneficial:
–– ↑↑ demand
demand oror value
value plants
plants
–– Plants
Plants are
are slow
slow or
or difficult
difficult to
to propagate
propagate
–– Endangered
Endangered species
species
•
Micropropagation
Advantages:
z Germplasm preservation
z
z Germplasm: the DNA of a species.
z
z In the past: seeds
• limited shelf-life
• require considerable storage space
• don’t preserve uniform characteristic
(variablility)
Micropropagation
Advantages:
z
z Any tissue or cell type can be used for
micropropagation
• Meristematic
Meristematic Tissue
Tissue
• Parenchyma Tissue
• Adventitious growth
• Virtually any plant cell
Principles of Micropropagation
Terms-
z Meristematic cells
– undifferentiated cells at shoot and root tips.
– greatest potential to produce cells that will become shoots or
roots.
z Parenchyma cells
– thin-walled cells that make up the bulk of most nonwoody
structures.
– can be induced to divide and differentiate.
Forms of Micropropagation
Callus
shoots initiating
from callus
Forms of Micropropagation
2. Microcuttings:
z
z The explant has buds (produce shoots directly-no callus).
z
z Shoots are separated from the explant and rooted.
root initiation
z Four stages:
I. Explant establishment (microshoot initiation)
II. Multiplication of microshoots
III. Rooting
IV. Acclimatization (hardening-off)
Micropropagation - The Basics
z Stage I – Explant establishment/initiation
1. Select explant.
2. Disinfestation (surface-sterilization).
3. Place explant in media.
–– Provides:
Provides:
•• Nutrients
Nutrients
•• Vitamins
Vitamins
•• Hormones
Hormones
fungal contamination
Micropropagation - The Basics
z Stage II – Multiplication of shoots
“Subcultured” or “transferred” to media with
↑[cytokinin] = shoot initiation
callus
[auxin] 1 1 1 10
[cytokinin] 1 10 30 1
Micropropagation - The Basics
z Stage IV– Acclimatization or Hardening off
Hardening off :
- transplanting the plantlet into soilless media with mist
- placing a clear plastic cover over the plantlet
Micropropagation - Summary
Explant placed in growth
Explant harvested media containing nutrients
and plant hormones
New plantlets
develop