Module 10: Micropropagation Flashcards
What is plant tissue culture?
Growth of plant tissue in sterile nutrient media in small containers using aseptic techniques
What is micropropagation?
The production of new plants through tissue culture
Cloning
What is an explant?
Small portion of tissue from the donor plant that is used to produce a new plant (any plant parts with meristems)
What are the applications of tissue culture?
Production of natural materials
Genetic improvement of crops
Production of plants with no disease
Quick multiplication (micropropagation)
- Year round
- Reducing number of stock plants and propagation area
What are the limitations of micropropagation?
Initial capital investment
Additional salary costs which are 75-80% of total production costs
Not commercially successful with some species
What steps do you take to prepare for explanting?
Mother plant needs to be true to type and disease free
Sterilization is required to ensure removal of microorganisms
What is the 1st stage of micropropagation?
Establishment and survival of the explant
Production of callus tissue
What is the 2nd stage of micropropagation?
Shoot growth where tissues are transferred onto media with low auxin/cytokinin ratio
What is the 3rd stage of micropropagation?
Root growth where plants are transferred onto media with high auxin/cytokinin ratio
What is the 4th stage of micropropagation?
Acclimatization where plants are transplanted into soil and adapted to greenhouse conditions
When can multiplication occur?
In each of the first three stages
What are some laboratory facilities involved in micropropagation?
Media preparation room
Transfer room
Culture/growing room
Anteroom
What happens in the media preparation room?
Holds all the ingredients needed to prepare the nutrient medium
(Distilled water, refrigerators for stock solutions, sinks, dishwasher, pH meter, balance/weighing tools, glassware, autoclave @ 121C @ 15 psi for 15-20 mins)
What happens in the transfer room?
Laminar flow hood with a HEPA filter to prevent any tissue culture contamination
Sterilizer to sterilize tools
Aseptic techniques (no talking to reduce air currents, minimizing movement, HEPA filter blows air at technician to reduce contamination, PPE)
What happens in the culture/growing room?
Maintain cultures under optimal environmental conditions
What happens in the anteroom
For staff to clean up and change into lab clothes
What are some media ingredients required in micropropagation?
Inorganic salts (no Carbon)
Sucrose
Glucose
Vitamins
Amino acids
Hormones
Agar
What documentation is required for micropropagation?
Media preparation sheet
Production sheet
Plant transfer sheet
