Presentation at the March 2013 dialogue workshop of the Biosciences for Farming in Africa media fellowship programme in Accra, Ghana. Please see www.sti4d.com/b4fa for more information

1. 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.

4. 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

5. 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

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

8. 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

9. 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

10. 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

11. 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.

12. 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

13. 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.

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

15. 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

16. 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.

17. 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

19. 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