Transcript In vitro - Bangor University

Assuring Reduced Variation in
Commercial Crops
(In vitro mass propagation experiences from Barbados and
Caribbean plant tissue culture laboratories)
Collin Scantlebury
University of the West Indies
Cave Hill Campus
BARBADOS
E-mail: [email protected]
Plants as Providers of Fine Chemicals, Conference presentation, 29-30 August 2012, School of Chemistry, Bangor, Wales
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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
“Plant tissue culture and its applications in plant biotechnology”
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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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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Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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- 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
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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Going bananas on the
banana plantations
- Annual threat of hurricanes
- 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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Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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Multiplying banana cultures
In vitro mass
propagation process
Weaned and hardened
vitro plants ready for field
planting
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)
• auxins
• cytokinins
IAA
BAP
Kinetin
TDZ thiaduzuron
IBA
NAA
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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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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Anther culture for polyploids
Somatic embryos for artificial seed
Other Plant TC Applications for Phyto-chemistry
Cell and callus cultures for induced variation
Cell culture for genetic modification
Cell culture for more ready access to target chemicals
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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Safeguards for Reduced Variation
Each 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/ephemeral
phenotypic 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)
Plants as Providers of Fine Chemicals, 29-30 August 2012, School of Chemistry, Bangor, Wales
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