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Assuring Reduced Variation in Commercial Crops
(In vitro mass propagation experiences from Barbados and Caribbean plant tissue culture laboratories)
Plants as Providers of Fine Chemicals, Conference presentation, 29-30 August 2012, School of Chemistry, Bangor, Wales
Collin ScantleburyUniversity of the West IndiesCave Hill CampusBARBADOSE-mail: [email protected]
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• UAPS – Unit for Advanced Propagation Systems, Wye College, University of London at Wye near Ashford, Kent, UK
• BDD – British Development Division• ODA – Overseas Development
Administration
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
“Plant tissue culture and its applications in plant biotechnology”
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Applications of plant tissue culture to phytochemistry
• Via in vitro mass clonal propagation each single plant cell potentially can generate an entire new plant and thousands of clones
• Introduction of genetic/phenotypic variability in tissue cultured plants and associated different chemicals
• Direct gene cloning and host cell modification to increase access to target chemicals (molecular biology techniques)
• Concerted precautions to ensure clonal fidelity
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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Caribbean Agricultural Research and Development Institute; University of the West Indies, Cave Hill Campus, St. Michael, Barbados www.cardi.org/. (Diamond Valley, St. Philip, lab)
Ministry of Agriculture. Graeme Hall, Christ Church, Barbados. www.agriculture.gov.bb. (Home Tissue Culture Laboratory, The Home Agricultural Station , St. Philip).
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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5Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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6Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
- A single plant from a remote forest
- Limited numbers in germplasm or genebank
- Research (field and laboratory studies) driven identification of a particular superior
plant species or variety (the athlete plant)
- Insufficient numbers for scientific field studies with replicated plots
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7Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
- Annual threat of hurricanes
Going bananas on the banana plantations
- Thousands of suckers required 875-1300/acre (2200-3300/hectare)
- Only sword suckers for planting (uniform)
- Uniformly growing and fruiting plants for better forecasting of harvesting
- Not seed-propagated like corn, wheat etc.
- Crop comprises a single variety not mixtures
- Black Sigatoka disease limits movement of germplasm across international borders
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8Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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In vitro mass propagation process
Weaned and hardened vitro plants ready for field planting
Multiplying banana cultures
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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macro & micro nutrients, vitamins, amino acids, sucrose, plant hormones, phytagel/agar, deionized H2O, HCl, NaOH,
(ethanol, NaOCl, fungicides, detergent/tween surfactant)
• cytokinins • auxins
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
IAA
IBA
NAA
BAP
Kinetin
TDZ - thiaduzuron
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Swelling and greening of meristematic buds within first week of explant initiation
Single shoot formation form (meristematic) bud or eyes
Single shoot ready for subculture or rooting (or splitting down the middle)
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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12Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
Other Plant TC Applications for Phyto-chemistry
Cell and callus cultures for induced variationCell culture for genetic modificationCell culture for more ready access to target chemicals
Anther culture for polyploids
Somatic embryos for artificial seed
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13Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
Safeguards for Reduced VariationEach explant can produce thousands of offshoots but be conservative
In vitro multiplications rate may be variable so don’t stress the system
Use judicious levels of plant hormone (cytokinin, auxin, gibberillin)
Start clonal propagation process with proven mother plant (athlete plants)
Require field studies to ensure true- to- type-ness and not epigenetic/ephemeralphenotypic characteristic or environmental effects being observed
Field surveys of vitroplants are required to verify frequency of off-types/sports
Use preformed meristems (nodal and shoot tip explants) rather than leaf, stems,anthers etc. which involve callus stages
Selection of weaned and hardened plants (size, production dates, iso-lining)