This document discusses genetically modified crops and their potential environmental impacts. It describes current and future GM crop traits such as insect resistance, herbicide tolerance, stress tolerance, and production of pharmaceuticals in plants. Major concerns discussed include effects on non-target species, gene flow and transgene escape. The document outlines EPA research on monitoring these impacts, including assessing non-target effects using molecular techniques to detect gene expression changes in species exposed to GM crops. It provides examples of monitoring bentgrass for transgene escape and using sentinel plants and resident populations to track gene flow. The overall goals are to apply molecular monitoring to at-risk species and ensure the safety of biotech crops.

1. AssignmentAssignment
Subject : Breeding Designer Crops (GPB822)Subject : Breeding Designer Crops (GPB822)
Presented by: Mr.Presented by: Mr. IndranilIndranil BhattacharjeeBhattacharjee
Student I.D. No.: 17PHGPB102Student I.D. No.: 17PHGPB102
Presented to : Prof. (Dr.)Presented to : Prof. (Dr.) B.G.SureshB.G.Suresh
SamSam HigginbottomHigginbottom University of Agriculture, Technology & SciencesUniversity of Agriculture, Technology & Sciences
AllahabadAllahabad--211007211007

2. Agricultural Biotechnology:
Genetically modified crops

3. 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

4. Major environmental and health
concerns regarding GM crops
• Non-target effects
• Gene flow /
Transgene Escape
• Insect Resistance
• Allergenicity

5. 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.

6. 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

7. Agricultural Pesticides:
UnintendedUnintended
EnvironmentalEnvironmental
ImpactImpact
Intended and unintended effects
Human HealthHuman Health
EffectsEffectsTargetedTargeted
InsectInsect
PestPest

8. 80 million acres of corn planted per year
(20% of total crop acreage)
Western Corn
Rootworm (WCR) is
major pest of corn.
Conventional insecticides
applied to 14-18 million
acres per year.
WCR responsible for 1
out of 7 insecticide
applications for all
agricultural crops
WCRWCR
damagedamage

9. 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

10. BtBt--corncorn
NonNon
BtBt--corncorn
NonNon
BtBt--corncorn
Bt-corn reduces the need for
conventional pesticides:

11. The Evolution of Bt-Resistance is an
Environmental Problem
????

12. Current Research
• 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

13. 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

14. 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

15. Non-Target Effects: The Agricultural
Landscape
?
?
?
NonNon--economiceconomic
EconomicEconomic

16. 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

17. 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

18. Bt CornBt Corn
WCRWCR
Gene Expression ChangesGene Expression Changes
DIET +ROOT +BTDIET +ROOT +BT
STDDIETSTDDIET
1 3 4 5 62
bt
DIET+ROOTDIET+ROOT

19. 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
RelativeGene
Expression
cnt
bt
Results from QPCR for six clones
identified through differential display.

20. 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

21. 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

22. Gene Flow / Transgene Escape

23. Major Issues of Transgene
Escape / Gene Flow
• Contamination of food supply
• Creation of “super-weed” crops
• Transgene escape to native relatives
via hybridization, creation of super-
weeds
• Genetic pollution

24. Keys to Prevention of
Transgene Flow
• 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

25. Case Study of EPA work
on Transgene Flow
• Herbicide
(Roundup)
resistant bent
grass
• Considered for
turf management
• Improve golf
courses

27. CD = managed
bentgrass field
Sentinel plants =
potted plants
Resident = naturally
occurring

28. 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).

30. 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 by
monitoring potential environmental
problems.