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.
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.
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
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
Aseptic Transfer Area Laminar air flow Dissecting microscopes Dissection instruments Gas outlet Vacuum facility Sterilizer
Culture Room Environmentally controlled Incubators with controlled temperature Rotary shakers Lux meter Space for cultures requiring complete darkness
Analytical Room Colorimeter Low speed centrifuge Inverted centrifuge Chemical reagent racks Viscosity meter Gas outlet
Acclimatization Room High illumination(4,000-10,000 lux) High humidity(90-100% through mist & fog systems)
Miscellaneous Items Air conditioners Uninterrupted power supply Bunsen burners Aluminium foils Fluorescent lamps Fire fighting equipment
Media No single medium supports growth of all tissues. Some basic factors Callus induction Organogenesis Murashige-Skoog medium, White’s medium, woody plant medium