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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.

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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

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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

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Focused crops
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Food crops
•Cassava
•Plantain/
•Banana
•Yam
•Sweetpotato
Tree crops
•Shea tree
•Bamboo
Medicinal
Plants
•Phyllantus
•Cryptolepis
•Afromomum

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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

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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

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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

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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.

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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

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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.

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Incubation conditions
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 Photoperiod: 16/8 hours day/light
 Temperature: 25-28ºC
 Light intensity 3,500-4,500 lux
 High humidity

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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

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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.

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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

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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