Nhap mon cong nghe sinh hoc

Plant Tissue Culture
Chapter 17 & 18
Plant T.C.?
Technique used to maintain and propagate:
Plant cells
Tissues
Organs

To maintain and enhance characteristics

Requirements?
Asepsis
Nutrient media
Controlled environment
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.
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
Media constituents
(solid or liquid)
Inorganic salts
Sugar
Vitamins
Hormones
Gelling agents
water
Controlled environment
Light
Temperature
Humidity
Air
Wastes
Agricultural applications?
Rapid clonal propagation
Elimination of pathogens
Development of superior cultivars
Production of biochemicals
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
Process
Obtain Explant
Preparation of media
Isolate growing point
Transfer to medium
Explant
Piece of plant used to initiate a culture.
Media Preparation
Prep room
Inorganic salts
Organic compounds-hormones
Support media
Agar, Liquid
Facilities- Autoclave, pH meter, hot plate, various utensils, etc.
Isolate Growing Point
Transfer Hood-using laminar air flow through HEPA filters
Sterilization facilities
Bleach rinsing, sterile water
Microscope
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
Stages
I Initiation
II Multiplication
III Root Formation
IV Acclimatization to greenhouse environment
Risks involved
No guarantee on exclusion of pathogens
Cultivar traits may not be retained
Possibility of frequency of aberrants increased with each sub-culture
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.
Development of superior cultivars
Germplasm storage
Embryo rescue
Ovule and ovary cultures
Anther and pollen cultures
Screening cultured cells and protoplasts
Protoplasmic fusion
Future and Current Development of T.C.
Genetic engineering
Pathogen exclusion
Somatic cell embryos encapsulated into clonal seeds