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Part 2:Understanding Plant
Growth In Vitro
Richard R Williams
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1. Fundamental Concepts / History
2. Types of Cells or Tissues
3. Cell to Cell Interaction
4. Growth Processes
Topics to be covered:
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“… the aseptic culture of plant protoplasts, cells, tissues or organs under conditions which lead to cell multiplication or regeneration of organs or whole plants “
What is Plant Tissue Culture?
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Cell theory and totipotency
Callus formation and growth substances
Early cell culture
First plant tissue cultures
Animal tissue cultures
Plant organ culture
1 History
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Cork tissue as observed by Robert Hooke in 1664
Cell theory ….
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Totipotency ….
… each living cell has a complete genetic blueprint and therefore has the potential to develop into an entire plant.
… cells lines differentiate to form specialised tissues and organs
… unlike animal cells, living plant cells can de-differentiate and then re-differentiate to form different cell types
… complete genetic blue print
… cells differentiate
… living plant cells can re-differentiate
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Callus Formation ….…
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…Haberlandt .. early 1900’s … proposed concept of totipotency … cells cultured under right conditions
Callus cultured from tree cambium (Gautheret, Nobecourt, Whire in the 1930s.
… cells kept alive but did not develop
Early Cell Culture ….
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Cell enlargement … role of auxins
Cell division ... role of cytokinins
- dependent on discovery of “growth regulators”
Early tissue culture ….
Regeneration from tobacco pith ..
(Skoog and Miller) … interaction of auxin
and cytokinin gives differentiation.
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GA for growth of shoots
Aux + Cyt + sucrose > vascular development
Culture of ‘thin layers’ … many interacting factors eg pH
Further development …
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Carrot plants from root cells – Stewart in 1964
[Steeves & Sussex 1972]
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Tobacco plants from single cells – Vasil & Hilderbrandt 1965
[Steeves & Sussex 1972]
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Plant Organ Culture ….
Murashige and Skoog 1962 - mineral media
micropropagation
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Many different types of cells
Varying degrees of specialisation
- Meristematic- Embryonic- Reproductive
2 Types of Cell & Tissues
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Shoot ... apical, … axillary
Meristematic tissues ...
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leaf trace
procambium
tunicanew leaf
corpus
pithcortex
apical meristem
axillary meristem
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Shoot ... apical, … axillary
Leaf
Root
Meristematic tissues ...
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Shoot ... apical, … axillary
Leaf
Root
Adventitious
Cambial tissues
Meristematic tissues ...
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Embryonic tissues ...
Pre-formed plant
Generative / somatic
Juvenile
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Reproductive tissues ...
Diploid / Haploid
Female / Male
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Monocots versus Dicots
‘Stems’ are leaf bases … not buds
Shoot apex at base
Intercallary leaf growth
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Important in intact plant
Isolated cell not function the same
Establishment of polarity
Expression of somaclonal (epigenetic)
variation
3 Cell to Cell Interaction
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Multiplication Apical dominance Differentiation
Growth Cell division Cell enlargement
Phasic development Juvenility Dormancy
4 Growth Processes
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Photosynthesis
Transpiration and water uptake
Cytological instability
4 Other Processes
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Physiological mechanisms Physical manipulation Chemical Plant Growth Regulators
(PGRs)
Environment
Genotype
5 Controlling Growth & Development
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Hormone theory Auxins Cytokinins
Gibberelins (GA) Ethylene Abscisic Acid (ABA)
Other PGRs
6 Growth Regulators
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Hormone BalanceAuxin Cytokinin
High Low
Low High
Root formation on cuttings Embryogenesis
Adventitious root formation in callus Callus initiation
Adventitious shoot formation Axillary shoot growth
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Auxin : Cytokinin ratio
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Hormone Action …..
Application + uptake
Endogenous + applied
Accumulation & Habituation
Interactions / Sequence
Pulsing vs prolonged exposure
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END of Part 2