uwe stolz nerl postdoctoral research fellow genomics workshop, chicago, il april 29, 2005
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Uwe Stolz NERL Postdoctoral Research Fellow Genomics Workshop, Chicago, IL April 29, 2005. Agricultural Biotechnology: Genomic Approaches to Monitoring the Effects of Genetically Modified Crops. Agricultural Biotechnology: Genetically modified crops. Current and Future GM Crop Traits. - PowerPoint PPT PresentationTRANSCRIPT
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Uwe StolzUwe Stolz
NERL Postdoctoral Research FellowNERL Postdoctoral Research Fellow
Genomics Workshop, Chicago, IL Genomics Workshop, Chicago, IL
April 29, 2005 April 29, 2005
Agricultural Biotechnology: Agricultural Biotechnology: Genomic Approaches to Monitoring Genomic Approaches to Monitoring the Effects of Genetically Modified the Effects of Genetically Modified
CropsCrops
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Agricultural Biotechnology:Genetically modified crops
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Current and Future GM Crop Traits
1. Insect Resistance (Plant Incorporated Protectants) – ex: BT corn & cotton
2. Herbicide Tolerance – ex: Glyphosate Resistant Corn & Soybeans (i.e. Roundup Ready)
3. “Value Added” Crops – ex: Golden Rice containing vitamin A
4. Stress Tolerance – ex: drought, salt resistant varieties
5. “Biopharming” – Production of drugs, chemicals on agricultural scales
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Major environmental and health concerns regarding GM crops
• Non-target effects
• Gene flow / Transgene Escape
• Insect Resistance
• Allergenicity
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Issues for Regional Monitoring Efforts
• Non-target Effects Species, Communities, Ecosystems
• Beneficial species, threatened / endangered species
• Gene Flow / Transgene Escape Crop to Crop, Crop to Wild Relative, Crop
to distant relative• Insect resistance genes, herbicide tolerance
genes
• Future – vaccines, chemicals, drugs, etc.
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Non-target effects of GM corn
1. Bt corn and non-target effects: Yieldgard Corn
2. ORD/NERL research approach / rationale
3. Research progress
4. Monitoring Applications
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Agricultural Pesticides:
UnintendedUnintendedEnvironmentalEnvironmental
ImpactImpact
Intended and unintended effects
Human HealthHuman Health EffectsEffectsTargetedTargeted
InsectInsect
PestPest
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80 million acres of corn planted per year(20% of total crop acreage)
Western Corn Rootworm (WCR) is major pest of corn.
Conventional insecticidesapplied to 14-18 million
acres per year.
WCR responsible for 1 out of 7 insecticide applications for all agricultural crops
WCRWCRdamagedamage
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CRW will continue to be a problem…Infested acres of CRW expected to increase: 39 million acres by 2013
EPA identified 10 insecticides used in agriculture as the most toxic to birds - 3 currently used to control corn rootworm (carbofuran, phorate and methyl parathion).
CRW have adapted to crop rotation with soybeans and have evolved resistance to several chemical insecticides
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Bt-cornBt-corn
Non Non Bt-cornBt-corn
Non Non Bt-cornBt-corn
Bt-corn reduces the need for conventional pesticides:
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The Evolution of Bt-Resistance is an Environmental Problem
????
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Current Research of ORD -NERL in Cincinnati
• Monitor resistance genes in pest populations-Identify resistance/tolerance genes
-Develop molecular techniques for rapid and cost effective screening
• Monitor non-target species for effects of Bt exposure
-Identify gene expression markers for Bt exposure in target pests
-Develop assays to test for effects in related non-targets
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Complements ecological monitoring by adding another level of analysis and increases the amount of information that can be gathered about the status and health of species
Advantages of Genetic Monitoring
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Non-Target-Monitoring
• What is a non-target species?
• How do we choose non-targets?
• Molecular genetics to monitor non-targets Gene Expression
• Benefits of molecular genetic techniques
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Non-Target Effects: The Agricultural Landscape
?
?
?
Non-economicNon-economic
EconomicEconomic
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Non-Targets
• Economic non-targets Non-target pests Beneficial predators Other beneficial Insects
• Non-Economic non-targets Threatened/endangered species Species of public concern Species whose primary habitat is not
agricultural
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Gene Expression
• Detects exposure to pesticides/PIP• Detects biologically meaningful exposure• Detects non-acute effects• Permits study of non-lab species• Highly sensitive - ex: EDCs in streams
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Bt CornBt Corn
WCRWCR
Gene Expression ChangesGene Expression Changes
DIET +ROOT +BTDIET +ROOT +BT
ST
D D
IET
ST
D D
IET
1 3 4 5 62
btDIE
T +
RO
OT
DIE
T +
RO
OT
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Effect of Bt-corn diet on gene expression
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
7A(1.21) 7D(4.23) 4A4(6.85) 4B1(6.18) 6A1(2.82) 6A2(1.82)
clone name
Rel
ativ
e G
ene
Exp
ress
ion
cnt
bt
Results from QPCR for six clones identified through differential display.
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Once Gene Expression Markers Confirmed
•Non-target species can be monitored for exposure to PIPs and/or pesticides (QPCR, RT-PCR)
•Several closely related species can be monitored at the same time to detect ecosystem level patterns
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Proof of Concept Research
•We are examining ground beetle species across corn growing areas•Examine several species communities•Look at exposure to Bt corn
-ELISA assays using beetle guts to check for presence of Bt toxin-Gene expression assays
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Gene Flow / Transgene Escape
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• Contamination of food supply• Creation of “super-weed” crops• Transgene escape to native relatives
via hybridization, creation of super-weeds
• Genetic pollution
Major Issues of Transgene Escape / Gene Flow
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• Use GM crops with few wild relatives (ex: Corn in the USA, Canada, Europe)
• Use GM crops with no weedy relatives (ex: Sorghum and Johnson Grass)
• Provide sufficient physical and physiological barriers to gene flow (ex: plant isolated patches, have appropriate borders, sterility)
• Monitor for the escape and movement of transgenes
Keys to Prevention of Transgene Flow
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Case Study of EPA work on Transgene Flow
• Herbicide (Roundup) resistant bent grass
• Considered for turf management
• Improve golf courses
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CD = managed bentgrass field
Sentinel plants = potted plants
Resident = naturally occurring
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In addition to PCR, also used Elisa (checks for protein product of transgene) and used seedlings from sentinel plants to check for RoundUp resistance (phenotype of transgene).
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Ultimate Goals
• Apply molecular monitoring methods to conserved, threatened, and endangered species (i.e. difficult to study in the lab).• Apply strategy to species / populations /
communities that are most at risk to changing crop practices.• Ensure the safety of biotech crops bymonitoring potential environmental problems.
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Acknowledgements
Ecological Exposure Research Division/Molecular Ecology Research Branch
Mark BagleyJim Lazorchak
Sobran Inc.
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