Plant Tissue Culture

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Plant Tissue Culture

• Chapter 17 18

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Plant T.C.?

• Technique used to maintain and propagate
• Plant cells
• Tissues
• Organs
• To maintain and enhance characteristics

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Requirements?

• Asepsis
• Nutrient media
• Controlled environment

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Asepsis

• Fundamental characteristic of plant tissue
  culture.
• Exclusion of superficially associated non
  infecting microbes.
• Other methods are required to eliminate viruses,
  viroids, internal bacteria, etc.

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Asepsis achieved by

• Donor organs treated with disinfestant.
• Bleach
• Detergents
• Alcohol
• Culture media sterilized by autoclaving.
• All work done in sterile, clean environment.
• Transfer hoods using HEPA filters

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Media constituents(solid or liquid)

• Inorganic salts
• Sugar
• Vitamins
• Hormones
• Gelling agents
• water

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Controlled environment

• Light
• Temperature
• Humidity
• Air
• Wastes

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Agricultural applications?

• Rapid clonal propagation
• Elimination of pathogens
• Development of superior cultivars
• Production of biochemicals

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Main objective

• Reproduce uniform plants via rapid clonal
  propagation using three systems
• Adventitious shoot formation
• Enhancing axillary branching of shoot tips
• Rooting of shoots
• Somatic cell embryogenesis

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Process

• Obtain Explant
• Preparation of media
• Isolate growing point
• Transfer to medium

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Explant

• Piece of plant used to initiate a culture.

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Media Preparation

• Prep room
• Inorganic salts
• Organic compounds-hormones
• Support media
• Agar, Liquid
• Facilities- Autoclave, pH meter, hot plate,
  various utensils, etc.

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Isolate Growing Point

• Transfer Hood-using laminar air flow through HEPA
  filters
• Sterilization facilities
• Bleach rinsing, sterile water
• Microscope

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Transfer Process

• Procedure done under transfer hood following
  procedures to maintain asepsis.
• Tissue isolated and removed
• Multiple rinse steps
• Tissue placed in tube/flask on to medium
• Vessel is covered, then placed under grow lights

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Stages

• I Initiation
• II Multiplication
• III Root Formation
• IV Acclimatization to greenhouse environment

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Risks involved

• No guarantee on exclusion of pathogens
• Cultivar traits may not be retained
• Possibility of frequency of aberrants increased
  with each sub-culture

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Elimination of pathogens

• Restores potential of plant for increased yields
  and/or quality.
• Save diseased plant from extinction.
• International exchange of propagules without risk
  of disease and insect spread.

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Development of superior cultivars

• Germplasm storage
• Embryo rescue
• Ovule and ovary cultures
• Anther and pollen cultures
• Screening cultured cells and protoplasts
• Protoplasmic fusion

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Future and Current Development of T.C.

• Genetic engineering
• Pathogen exclusion
• Somatic cell embryos encapsulated into clonal
  seeds

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