b4fa 2013 ghana: pineapple tissue culture - kenneth danso
TRANSCRIPT
Pineapple Tissue Culture
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K.E. Danso Biotechnology and Nuclear Agriculture Research Institute,
P. O. Box LG 80, Legon, Accra Ghana
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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
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
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