Pineapple Tissue Culture

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K.E. Danso Biotechnology and Nuclear Agriculture Research Institute,

P. O. Box LG 80, Legon, Accra Ghana

kaedanso@hotmail.com

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BNARI’S BIOTECHNOLOGY EXPERIENCE

Mission BNARI exists to carry out research and development activities on safe applications of biotechnology and nuclear science and transfer these technologies to end-users in order to enhance agricultural productivity, health delivery and industrialization.

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Vision Leading public institution that providing solutions to challenges in agriculture, health and industry through scientific knowledge in biotechnology and nuclear science.

Biotechnology and Nuclear Agriculture Research Institute

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Nuclear Agriculture Centre (NAC)

Biotechnology Centre (BTC)

Radiation Entomology and Pest Management Centre (REPMC)

Radiation Technology Centre (RTC)

Technology Transfer Centre (TTC)

Research Centres

Research activities

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Mutation induction Conventional hybridisation Genetic transformation Improving regeneration efficiency in food, tree and medicinal plants Water use efficiency Soil management studies Medical sterilisation Food preservation and extension of shelve life Control of pest and diseases using SIT

Other activities Training of students Farmers

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TRAINING OF STUDENTS AND INTERNATIONAL FELLOWS

Scientists from Africa at a practical section

Focused crops

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Food crops •Cassava •Plantain/ •Banana •Yam •Sweetpotato

Tree crops •Shea tree •Bamboo

Medicinal Plants •Phyllantus •Cryptolepis •Afromomum

Limitations of conventional propagation

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Pineapple (Ananas comosus L. Merr.) is vegetatively propagated

Have long propagation cycle about 18 months

Unexpected or sporadic natural flowering

Seeds cannot be used for propagation

Often associated with systemic viral, fungal and bacterial diseases

Mature at different times because they are not uniform

Why pineapple tissue culture?

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Rapid multiplication All year round production Disease elimination Production of uniform planting materials Germplasm conservation and exchange of plant genetic resources Prerequisite for genetic modification Enhance improvement of the crop

Methodology

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Explants for culture

slips, suckers, crowns ratoons leaves

Crowns are the preferred planting material since they have the potential to develop better root systems in some countries. For tissue culture slips are the most preferred

Explant Selection

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The planting (explant) to be cultured is usually taken from a healthy and vigorously growing ideally from the glasshouse. suckers or buds at the base of the leaves

slips found at the base of the fruit.

Caution: Buds must not be opened at the time of collection due to high microbial load on the explant.

Sterilization of explants

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Explants

Running tap water

Wash with sterile distilled water

Transfer to Flow chamber

Trim explants

Immerse with sodium hypochlorite Or 0.1% mercuric chloride

Rinse three times in SDW

Initiate on a pineapple culture medium

Culture medium

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Murashige and Skoog (1962) basal salt

30 g/l sucrose

100 mg/l myo-inositol

4.5 mg/l BAP

0.75 mg/l NAA

Gamborg B5 vitamins pH 5.8 autoclave 3.5 mg/l phytagel autoclaving at 121ºC for 15 minutes

Culture medium may be solid or liquid. Generally liquid medium enhances the production of more plantlets than solid but liable to contamination.

Incubation conditions

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Photoperiod: 16/8 hours day/light

Temperature: 25-28ºC

Light intensity 3,500-4,500 lux

High humidity

Subculture

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Splitting them into two equal halves

Each half is transferred onto fresh MS medium supplemented with BAP and NAA but at slightly lower concentrations than he initiation medium.

Well developed shoots without roots are transferred onto the same medium for proliferation

Rooting

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MS medium with lower concentration of BAP

relatively higher concentration of NAA or IBA or a combination of NAA and IBA.

Root development occurs within 4-8 weeks depending on the auxin in the culture medium.

Acclimatisation

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Shoots with roots are transferred to the greenhouse or the plant barn Gently remove plantlets from the culture vessels Washed off any phytagel adhering to roots Transfer to loamy soil mixed with cow dung or coconut husk (vermiculite or jeffy peat pellets can be used) Water of MS solution Cover with Watson module/plastic cup to create high humidity. Plastic cups are removed after four

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Acclimatisation

Field transfer

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Successfully weaned plantlets are transplanted on the field Plantlets will do well depending proper agronomic practices

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BNARI’S Role in the pineapple Industry

Significant role in the change over from growing smoot cayene and sugar loaf to MD2 at the international market Supplying of planting materials to individual farmers And some companies outside Ghana

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Thank you