Bài giảng Plant tissue culture - Chapter 17+18

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