SHOOT TIP CULTURE

1. Shoot Tip Culture

2. What is Shoot Tip Culture?
• Shoot tip culture may be described as the culture of terminal (0.1-1.0 mm)
portion of a shoot comprising the meristem (0.05-0.1 mm) together with
primordial and developing leaves and adjacent stem tissue.

3. Why shoot tip or meristem??
• Virus free plants
• No cell differentiation
• Virus cant move from cell to cell
• Intense metabolic activity
• No vascular system

4. PROTOCOL
• Protocol Remove the young twigs from a healthy plant. Cut the tip (1 cm) portion
of the twig.
• Surface sterilize the shoot apices by incubation in a sodium hypochlorite solution
(1% available chlorine) for 10 minutes.
• The ex- plants are thoroughly rinsed 4 times in sterile distilled water.
• Transfer each explants to a sterilized petri dish.
• Remove the outer leaves from each shoot
• After the removal of all outer leaves, the apex is exposed.
• Cut off the ultimate apex with the help of scalpel and transfer only those less
than 1 mm in length to the surface of the agar medium.
• Incubate the culture under 16hrs light at 25°C

5. STAGESOF SHOOT TIP CULTURE
• Murashige reported that there are three stages of culture:
Stage 1 is the culture establishment stage
• when explant may develop into single shoot or multiple shoots.
• At this stage explant are supplements with cytokinin like BA, kinetin and 2iP.
In stage 2
• the objective is to multiply the propagule and for this axillary shoot proliferation
is followed as it maintains higher genetic stability.
The stage 3
• purpose is regeneration of adventitious roots from the shoots obtain in stage 2
• Numerous studies have indicated that NAA is followed by IBA,IAA, 2,4- D and
other auxins are used for induction of root generation

8. Factors affecting shoot tip culture
• Size of ex plant
• Physiological condition of ex plant
• Season of culture
• Culture medium
• Storage condition

9. Application of Shoot-tip or Meristem Culture
1. Virus Elimination
• Plants are often infected with more than one type of virus, including some even
not known.
• A general term virus- free is used by commercial horticulturist for plants freed of
any type of virus

10. 2. MicroPropagation
• A sexual (seed) or vegetative propagation (vegetative e part) of whole plants
using tissue culture techniques referred to as micro propagation.

11. 3. Storageof GeneticResources
• Many plants produce seeds that are highly heterozygous in nature or that is
recalcitrant.
• Such seeds are not accepted for storing genetic resources. So , the meristem
from such plants can be stored in vitro.

12. 4. Quarantine
• Plantlets derived from shoot-tip or meristem cultures are easily accepted by the
quarantine authority for international exchange without any checking.
• Therefore, using this technique , crop plants can be easily exchanged in crop
improvement programmes that are based on materials from different parts of
the world.

13. CONCLUSION
• It is very effective method of cloning of plant material and to develop disease
free clean plant stock.
• Shoot Tip Culture is a part of plant tissue culture which is a sun-rise
technology and working as a catalyst of agricultural and industrial
development