INTERNATIONAL ISLAMIC UNIVERSITY ISLAMABADTopic of presentation: SHOOT TIP CULTURE

Presented by: Anum Yousaf 489-FBAS/BSBT/F13 Atia Gulzar 407-FBAS/BSBT/F13 Nosheen Rehman 436-FBAS/BSBT/F13

Presented to: MADAM RUBINA

Dated: 28th November 2016

Course: Agriculture Biotechnology

INTERNATIONAL ISLAMIC UNIVERSITY ISLAMABAD

CONTENT LIST• Introduction• What Is Meristem

Culture?• History Of Plant Tissue

Culture• Principle• Why Shoot Tip or

Meristem??• Protocol And Stages Of

Shoot Tip Culture• Factors Affecting Shoot

Tip Culture• Application Of Shoot Tip

Culture• Limitations • Conclusion

INTRODUCTIONShoot 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.

Why shoot tip or meristem??

Virus free plantsNo cell differentiationVirus cant move from cell

to cellIntense metabolic activityNo vascular system

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 incuba tion in a sodium hypochlorite solution (1% available chlorine) for 10 minutes. The ex- plants are thoroughly rinsed 4 times in ster ile distilled water.

Transfer each explants to a sterilized petri dish.

Cut off the ultimate apex with the help of scalpel and transfer only those less than 1 mm in length to the sur face 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

STAGES OF SHOOT TIP CULTUREMurashige 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.

STAGES OF SHOOT TIP CULTURE

The stage 3 purpose is regeneration of adventitious roots from the shoots obtain in stage 2Numerous studies have indicated that NAA is followed by IBA,IAA, 2,4-D and other auxins are used for induction of root generation.

STAGES OF SHOOT TIP CULTURE

Factors affecting shoot tip culture Size of ex plant Physiological

condition of ex plant

Season of culture Culture medium Storage condition

Application of Shoot-tip or Meristem Culture1.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

2. Micro Propagation• A sexual (seed) or vegetative

propagation (vegetative e part) of whole plants using tissue culture techniques referred to as micro propagation.

3. Storage of Genetic Resources

• 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.

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.

CONCLUSIONIt 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