TISSUE CULTURE ON BANANA SUCKERS AND RICE CALLUS (MICROPROPAGATION)

PLANT TISSUE CULTURE

MEMBERS :

•KAREN LENGGIAU ANK AUGUSTINE TUJOH (B11A124) SBS2•NUR AMALINA BINTI SHAHABUDIN (B11A294) SBS2•NURHAIZAN BINTI KAMARUZZAMAN (B11A323) SBS2•SITI SUBAILAH BINTI HAJI SUOD (B11A388) SBS2•WAN SHAHIRAH NERINI BINTI ROSLI (B11A423) SBS2 •MUHSIN BIN ABD RAZAK (B11A251) SBT2•MOHAMAD IQBAL BIN ALI AKBAR (B11A233) SBS2•LAM CHIN JACK (B11A143)SBT2

LECTURER : DR FATIMAH KAYATGROUP : 7 DATE OF PRESENTATION : 28TH MAY 2013

BANANA SUCKERS

INTRODUCTION

1)Providing us a basic understanding of physical and chemical requirements of cell, tissue, organ culture, their growth and development.

2)To learn and understand a procedure that is often used to propagate many plants of the same genetic background.

OBJECTIVES

Banana belongs to Musaceae family, second largest food-fruit crops of the world produced in the tropical and subtropical regions.several constraints including highly season dependent and sometimes poor quality of planting material of banana become a limiting factor for banana propogation. Therefore, tissue culture technology was considered an appropriate option to overcome this problem.Application of tissue culture technology to propagate banana plants gives some advantages resulting in disease free planting materials, more vigorous growth, high yields, better quality of fruits, earlier fruiting and more uniform crop since they are made from selected high yielding mother plants.

The suckers are the major source of planting material for banana and normally remain true-to-

type.

-sword suckers-water suckers

MEDIA PREPARATION

MS MEDIA

200 ml of distilled water is added to a 500 ml conical flask

15g of sugar

25ml of Macronutrients stock

2.5ml of Micronutrients stock

1ml of Vitamin stock

2.5 ml of Plant Regulator Growth stock

(Benzylaminopurine 5mg/L)

4g of agar is added

Distilled water is added to adjusted the volume to 500ml.

Natural regeneration of cultivated bananas through suckers is very slow

due to hormone-mediated apical dominance of the mother plant. So, as an alternative, shoot tips from young banana suckers with growing buds or

cut rhizomes called ‘bits’ and ‘peepers’ are most commonly used as explants

for accelerating the rate of in vitro propogation of banana.

MS MEDIA with 5mg BAP

IN-VITRO PROPAGATION (BANANA SUCKERS)

• The suckers of banana tree is trimmed by discarding the outer layers of the pseudostem and roots leaving only shoot tips containing rhizome tissue

• 70% bleach and Tween-20 for (15 mins)

spray the surface of laminar workflow with ethanol

The half trimmed banana suckers then sterile immediately by put in bottle contain 70% ethanol and shake it for 20 seconds and then 3

times of sterile distilled water.

Trimmed again

RESULTS AND DISCUSSION

CONTAMINATED!!

bacteria

fungi (mycelium)

WHY TISSUE CULTURE

• Multiplying plant in short time and in large quantity

• Disease free plants

• Save space for early plantation/germination thus reduce cost for land used

• Cultured specific/usefull part of plant

PROCESS

• Sucker selection (sword sucker)

• Sterilization technique

• Growth media (MS media)

• Culturing

• Sub-culture

FACTOR

• Germination

suitable nutrient composition (media)

duration of alcohol use in sterilization

• ContaminationCrowded work station

Media/explant sterilization

IMPROVEMENT

• Container

• Sealing/ cap

• Media

• Working area/ method

SUGGESTION•use shoot tips from young banana suckers with growing buds or cut rhizomes called ‘bits’ and ‘peepers’ are accelerating the rate of in vitro propogation of banana.

•Place the banana culture in complete darkness first for 1 week.

•add antioxidant in media such as ascorbis acid.

Callus Induction of

Rice (Oryza sativa)

Seeds

INTRODUCTION

Rice is the most important crop in Malaysia

because it is staple food for Malaysian

In Malaysia, rice demand is higher than

production rate

New technology must be developed to increase

the rice production

Tissue culture technology is one of the way to

produce high quality rice which is can produce

much rice grains, resistant to environmental

stress and shorter development stage ( cut

time so that can harvest quickly)

Importance of Tissue Culture in Rice production

* Callus can undergo mutation to achieve the desired traits

* The following methods are used to induce mutation in callus :

1. Physical mutagens (irradiation)

2. Chemical mutagens (carcinogens)

* The application of somaclonal variation in rice callus includes:

1. For crop improvement

2. Producing plants that resistant to environmental stress

3. To increase yield

4. To increase plant fertility

Description : ContaminatedFactor : Technique of handling

Description : ContaminatedFactor : Contamination from media

Formation of Callus- Compact callus

Unorganized, undifferentiated, growing and dividing

mass of cells.

Form from the wounding part.

Rapid plant multiplication for quality seed material.

Performed in the dark as light can encourage

differentiation of the callus

Important in plant biotechnology.

Manipulation of the auxin to cytokinin ratio in the

medium can lead to the development of shoots, roots or

somatic embryos from which whole plants can

subsequently be produced.

Callus

Callus Suspension culture

Place friable callus into a liquid medium and agitate

Single or small clumps of cells will be released into the

medium, where they keep growing and dividing to eventually a

cell suspension.

The size of inoculum have to be sufficient , suspension can be

build up faster.

Too much cells and there is a risk that toxic products from

damaged cells could accumulate to a inhibiting level.

Cell suspension cultures should also be subcultured

periodically.

CONCLUSION

It can be concluded dichlorophenoxyacetic acid 2, 4- D is suitable for callus induction of rice seed.The role of 2,4-D in cell division is to increase the rate of cell division and this attributes to the increased amount of callus.

SUGGESTIONREPEATED EXPERIMENTS.

For callus induction MS medium supplemented with different concentrations of 2,4-D

DIFFERENT VARIETIES OF RICE.

ASD 16, ADT 43, Basmati 370, Pusa Basmati and Pokkali

THANK YOU