Shoot tip culture

1. INTERNATIONAL ISLAMIC UNIVERSITY ISLAMABAD
Topic of presentation: SHOOTTIP CULTURE
Presented by: AnumYousaf 489-FBAS/BSBT/F13
AtiaGulzar 407-FBAS/BSBT/F13
NosheenRehman 436-FBAS/BSBT/F13
Presented to: MADAM RUBINA
Dated: 28th November 2016
Course: Agriculture Biotechnology
INTERNATIONAL ISLAMIC UNIVERSITY ISLAMABAD

2. CONTENT LIST
• Introduction
• What Is MeristemCulture?
• HistoryOf Plant Tissue Culture
• Principle
• Why Shoot Tip or Meristem??
• Protocol And Stages Of Shoot Tip Culture
• Factors Affecting Shoot Tip Culture
• ApplicationOf Shoot Tip Culture
• Limitations
• Conclusion

3. INTRODUCTION
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.

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

6. PROTOCOL

7. 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.

8. 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
Remove the outer leaves from each
shoot
After the removal of all outer
leaves, the apex is exposed.
Protocol

9. STAGESOF SHOOTTIP 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.

10. In stage 2 the objective is to
multiply the propagule and for
this axillary shoot proliferation
is followed as it maintains
higher genetic stability.
STAGESOF SHOOTTIP CULTURE

11. 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.
STAGESOF SHOOTTIP CULTURE

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

13. 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 like potato

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

15. 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.

16. 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.

17. 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