Documente Academic
Documente Profesional
Documente Cultură
Application
By;
Nasir Hussain
Tissue Culture
• Production of microbe-free plant material in
an in vitro conditions from any part of the
plant in an aseptic (sterile) environment, such
as sterilized nutrient medium.
• It is non conventional method of vegetative
propagation for any crop specie.
• Conventionally vegetative propagation take
place through cuttings, budding, grafting and
layering etc.
Tissue Culture Techniques
• Micropropagation .
• Production of pathogen free plants.
• Meristem Culture.
• Somatic embryogenesis.
• Organ Culture.
• Callus Culture.
• Embryo culture.
• Anther and pollen grain Culture
• Protoplast culture etc.
Advantages of Tissue Culture
• Elimination of viruses from infected plants
• Rapid multiplication of clones (elite)
• Vegetative propagation of difficult to
propagate species
• All the year round propagation of clones
• True to type plantlet production
• Rapid multiplication of seedlings (in cases
where seed is hard to get)
Applications of Tissue Culture
• Organ, meristem, • Micropropagation,
shoot. disease elimination,
germplasm preservation.
• Undifferentiated • In vitro selection,
(callus suspension). secondary metabolites
production
• Embryo rescue. • Wide hybridization
• Somatic • Artificial seeds, genetic
embryogenesis. engineering.
Applications of Tissue Culture
• Anther, microspore • Homozygous line
culture. production.
• Protoplast • Gene pool
diversification
Tissue culture methods for virus
eradication
Thermotherapy, chemotherapy and meristem
tip culture.
• Incubation of plants or of cultures at
relatively high temperatures (32° to 40°C)
either alone or
• in combination with the excision and culture
of very small stem tips is the only method of
ridding infected cultivars of their viruses.
• Antivirus (Ribavirin).
Tissue culture methods for virus
eradication
• Excise and culture the apical meristem of an infected
plant because this region is most likely to be free of
viruses.
• It involves a very delicate dissection and such small
explants are difficult to grow in culture.
• A very small stem tip which has, in addition to the
apical meristem, one or two leaf primordia. It is
much easier to grow in culture.
• High magnification stereomicroscope is also needed
to excise the meristems (0.1-0.5 mm).
Tissue culture methods for virus
eradication
• However, some viruses invade the
apical meristem and, in these cases, it
is necessary to apply heat treatments
to the plant. In the past, this has meant
heat treatment of the whole plant
followed by stem tip culture. An easier
method involves the application of the
heat treatment to aseptic cultures.
MERISTEM TIP CULTURE
Axillary
bud
Internode
Root
Micropropagation Methods
• Micro cuttings
• Small sections of plant
tissue are cut from the
mother plant
• Placed into media
• Grown out or
subdivided again to
produce more plants
Micropropagation Methods
• Shoot Tip Culture (less 10 mm)
• Meristem culture (0.1-0.5 mm)
Surface sterilize, wash & culture.
Higher cytokinins to over come
apical dominance.
Adventitious bud production.
sub-culture frequently.
Basic in vitro propagation ...
Micropropagation Stages
♣ in vitro micro-grafting
♣ Genetic conservation
♣ Plant improvement
♣ Experimental system