3. Introduction
Principles of plant tissue culture :
Tissue culture simply directs and assistants the
natural potential within the plant to put forth new
growth and the multiply in highly efficiently and
predictable way.
Totipotency i.e is the capacity of an individual
all to regenerate in to the whole plant, the
concept of totipotency (T.H. Morgan, 1901).
All the plant cells have their property since
potential lies mainly in cellular differentiation.
This indicates that all genes responsible for
differentiation tissue or organ are able to
express only under adequate culture conditions.
4. Introduction
The three main changes or stages is the complete
development an ordered change or progress often towards a
higher more complex state of a cell are
Cell division
Cell elongation
Cell Maturation
Two kinds of plant growth are possible in vitro
Organized Growth : Occurs either when organized plant parts or
organs such as the growing point apical Meristem of shoots or
roots leaf initials, young flower buds and small fruit are
transferred to culture (where they may continue to grow with their
structure preserved ) or when these structure are formed afresh
during the culture of unorganized tissues.
Unorganized Growth : Occurs when pieces of whole plant are
cultured in vitro. The tissue thus formed typically lack any
recognizable structure contain only limited no of the many
different kinds of specialized cells found in an intact plant.
5.
6. Basic Techniques
Setting up of a tissue culture lab
requires proper planning.
It is divided into 5 areas
Media preparation room
Aseptic transfer area
Culture room
Analytical room
Acclimatization room
7. Media Preparation Room
Refrigerator & freezer
Water purification & storage system
Glassware washing facility
Continuous supply of single & double
distilled water
Culture media, washing powder,
disinfectants
Cabinets or shelves
8. Aseptic Transfer Area
Laminar air flow
Dissecting microscopes
Dissection instruments
Gas outlet
Vacuum facility
Sterilizer
9. Culture Room
Environmentally controlled
Incubators with controlled temperature
Rotary shakers
Lux meter
Space for cultures requiring complete
darkness
10. Analytical Room
Colorimeter
Low speed centrifuge
Inverted centrifuge
Chemical reagent racks
Viscosity meter
Gas outlet
11. Acclimatization Room
High illumination(4,000-10,000 lux)
High humidity(90-100% through mist &
fog systems)
12. Miscellaneous Items
Air conditioners
Uninterrupted power supply
Bunsen burners
Aluminium foils
Fluorescent lamps
Fire fighting equipment
13.
14. Media
No single medium supports growth of all
tissues.
Some basic factors
Callus induction
Organogenesis
Murashige-Skoog medium, White’s
medium, woody plant medium
25. Sterilization of plant tissues
Plant tissues
Sodium hypochlorite (NaOCl): most
common to sterilize plant tissues
Calcium hypochlorite (CaOCl): less damage
than NaOCl
Hydrogen peroxide (H2O2): easily removed
from tissues
Other substances: bromine water, silver
nitrate, mercuric chloride
26. Cleaning
Glassware/plasticware in 10%
commercial detergent liquid.
Wash with tap water (to remove
detergent).
Rinse in double distilled water, and allow
to dry overnight.