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Understanding Biotechnology
Steve Strauss, Professor, OSU
Forest Science, Genetics, Molecular and Cellular Biology
Director, Outreach in Biotechnologyhttp://wwwdata.forestry.oregonstate.edu/orb/
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Outreach website
http://wwwdata.forestry.oregonstate.edu/orb/
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Educational activities Food for Thought Lecture Series / 2005-2008
Streaming video - OPAN/OPB usage
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The plan• What is biotechnology• GMOs
– State of usage in the world– How it works– The general concerns surrounding them
• Non-GMO biotechnologies (Dave Harry)– Genomics and DNA markers
• Break-outs for grass seed specifics– Commercialization issues, GMO testing, grass
industry biotechnologies
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What is biotechnology?Amer. Heritage Dictionary (2000)
• 1. The use of microorganisms or biological substances such as enzymes, to perform industrial processes.
• 2a. The application of the principles of engineering and technology to the life sciences; bioengineering.
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A more crop oriented definition of biotechnology
• Use of technologies that affect physiology, genetics, management, or propagation
• Most common uses– Microorganisms for fermentation of plant
products– Plant tissue culture for propagation– DNA sequencing and indexing for
identification (DNA fingerprinting)– Gene isolation, modification, and insertion
(genetic engineering, “modern biotechnology”)• GE, GEO or GM, GMO
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Why emphasize GE forms of biotechnology?
GE crops have been taken up rapidly by farmers when available, have had
large benefits, and have great economic and humanitarian potential
Exploding science of genomics fuels rapid discovery, innovation
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Rapid rise of GE crops in developed and developing world
http://www.isaaa.org
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Many social issues with major impacts on use / acceptance
• Few GMO crop types in production– Maize, soy, cotton, canola– Insect, herbicide tolerance traits– Small amounts of viral resistance (squash, papaya)
• Benefits of reduced tillage, reduced pesticide use, improved yields, reduced costs
• But other traits and crops mostly on hold– Substantial social resistance and obstacles to their use
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Defining GMOs
• GEO / GMO = creation of a “recombinant DNA modified organism”– It’s the method, can use native or foreign genes
• DNA isolated, changed/joined in a test tube, and re-inserted asexually– Vs. making crosses or random mutations in
conventional breeding
• Powerful breeding tool but can generally handle one to a few genes at a time – Simple traits can be designed, but without constraints
from native gene pools– That’s why its called genetic engineering, though we
are modifying, not building, a new organism
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Assembling a gene
Controls level of expression, and
Where and when expressed
Provides stability to messenger RNA, and
Guides processing into protein
Coding sequencePromoter Terminator
Can mix and match parts & can change sequences to improve properties
Protein
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Promoter (controls expression)
Gene (encodes protein)
Examples of promoter : gene combinations produced via recombinant DNA methods
Phenolic pathway enzyme (bacteria)35S-CAMV (plant virus)
RNA degrading enzyme (bacteria)Pollen sac (tobacco)
Herbicide tolerant
Male-sterile
FMV (plant virus) Insect toxin protein (bacteria)
Insect resistance
Oilseed (canola) Insulin (human)
Improved nutrition
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Recombinant DNA modification of native plant genes
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How are GE plants produced?
Step 1Getting whole plants back from cultured
cells = cloning
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Differentiation of new plant organs from single cells
Leaf-discs
First step is de-differentiation into “callus” after treatment with the plant hormone auxin
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Shoots, roots, or embryos produced from callus cells using plant hormones
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Step 2
Getting DNA into plant cells
Main methods- Agrobacterium tumefaciens- Biolistics [gene gun]
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Agrobacterium is a natural plant genetic engineer
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Agrobacterium gene insertion
Gene of interest
Agrobacterium tumefaciens
Engineeredplant cell
T-DNA
Ti Plasmid
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Only a few cells get modified so need to identify and enrich for the engineered cells
Not all cells are engineered, or engineered the same. Thus need to recover plants from that one cell so the new plant is not chimeric (i.e., not genetically variable within the organism)
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Hormones in plant tissue culturestimulate division from plant cells
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Antibiotics in plant tissue culturelimit growth to engineered cellsOther kinds of genes can also be used to favor transgenic cells
(e.g., sugar uptake, herbicide resistance)
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Transformation of bentgrass(Wang and Ge 2006)
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Glyphosate-tolerant FescueConventionally-bred Patented Varieties
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GE traits under development in forage and turfgrasses
Wang and Ge, In Vitro Cell Develop. Biol. 42, 1-18 (2006) • Nutritional quality
– Lignin reduction, increase of sulfur-rich proteins
• Abiotic stress tolerance– Drought, frost, salt
• Disease/pest management– Fungal, viral, herbicide tolerance
• Growth and nutrient use– Flowering time, phosporus uptake
• Hypoallergenic pollen• Bioethanol processability
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Problems and obstacles to wider use of GE crops
• Regulations complex, uncertain, changing, and very costly – Three agencies can be involved– Environmental and food/feed acceptability criteria
complex, stringent compared to all other forms of breeding
• Unresolved legal issues of gene spread, safety assessment, liability, marketing, and trade restrictions
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Legal actions• USDA sued over process for granting field
trial permit for GE bentgrass and GE biopharma crops
• USDA sued over deregulated Roundup- resistant alfalfa– First time an authorized crop forced to be
removed from market
• USDA required to do EIS for alfalfa, one was already underway for bentgrass
• Scotts fined $ 500K over Roundup Ready bentgrass field trial
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Strong and well funded political and legal resistance
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Intellectual property issues• New, costly, overlapping “utility patents” issued
for genes and crops since 1980• Patent “anticommons”
– Major costs, uncertainties for use of best technologies and usually need several licenses for an improved crop
• Major litigations ongoing for years to decades– Basic Agrobacterium gene transfer method– Bt insect resistance gene innovations
• Regulatory risks make large companies very reluctant to license to small companies, academics
• Public sector, small companies find it very hard to cope with the costs, obstacles
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• Strong polarization on benefits vs. risks– A highly vocal, concerned minority (~20%)
• A majority whose level of acceptance varies widely among applications depending on benefits and ethical views– Strong resistance to animal applications, and
to impacts that appear to harm biological diversity
• Very low knowledge of the science, technology
Varied public approval
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Rutgers survey data - USA (2005)http://www.foodpolicyinstitute.org/resultpub.php
http://www.foodpolicyinstitute.org/docs/reports/NationalStudy2003.pdf
• Seven in ten (70%) don't believe it is possible to transfer animal genes into plants
• Six in ten (60%) don't realize that ordinary tomatoes contain genes
• More than half (58%) believe that tomatoes modified with genes from a catfish would probably taste fishy
• Fewer than half (45%) understand that eating a genetically modified fruit would not cause their own genes to become modified
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Education needs: Gullibility
• "People seem to have a great number of misconceptions about the technology. As a result, they seem to be willing to believe just about anything they hear about GM foods.“
• Very few universities take an active role in outreach, education– University of California system an exception
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Summary
• GE is a method, not a product
• GE crops a major presence and with major science and technology push forward
• GE method highly regulated, causing great costs and uncertainties both for field research and commercial development
• Social/legal obstacles slowing or blocking investment outside of the major crops and large corporations
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Understanding Biotechnology
Part 2:
Genomics and DNA MarkersDavid Harry
Department of Forest ScienceAssoc. Director, Outreach in Biotechnology
http://wwwdata.forestry.oregonstate.edu/orb/
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DNA-based Biotechnologies
• Genetic engineering (GE, GMO)– direct intervention and manipulation– gene manipulation and insertion through an
asexual process
• Genomics & DNA markers – are generally descriptive, examining the
structure and function of genes and genomes– manipulating genes and genomes is indirect,
through selection and breeding
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Some definitions• Genes
– a piece of DNA (usually 100’s to 1000’s of bases long)– collected together along chromosomes– serves as a structural blueprint or a regulatory switch
• Genome– an entire complement of genetic material in the nucleus of an
individual (excluding mitochondria and chloroplasts) – genes, regulatory elements, non-coding regions, etc – tools for describing genomes include maps and sequence
• DNA marker– some type of discernable DNA variant (variation, or
polymorphism) that can be tracked– tracking the +/- of markers offers powerful tools for managing
breeding populations and, increasingly, for predicting offspring growth performance
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For today:• Basics of DNA markers• DNA markers & fingerprints
– are fixed for the life of an individual– can be used to identify individuals
• Marker inheritance (parent to offspring) – nuclear markers– parentage verification– genome mapping
• Associating markers and traits– maps and associations– marker breeding (MAS/MAB)
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Genes are located on packaging platforms called
chromosomes
Genomes, genes, and DNA
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DNA markers reveal subtle differences in DNA sequence
A
A
AT
T
C G
G
A
A
AT
T
C C
G
T
C
CA
C
G T
G
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CA
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G T
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G<>C
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Marker “1”
Marker “2”
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AgeAge
A DNA fingerprint is fixed throughout an individual’s
life
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MCW-305
MCW-184
MCW-087
DNA Fingerprints to Verify Identities 22 Paired Samples Collected at Different Times
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S DProgeny
Pedigree errors: “non-parental” marker types
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Genetic Map: Perennial Ryegrass
Gill et al. 2006
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X XThen, evaluate genetic makeup early to select
young birdsFirst, associate performance and
genetic makeup
X X
How might genetic markers accelerate breeding?
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a
b
c
d
e
f
g
h
i
j
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l
m
Hypothetical genes (QTLs) affecting economic traits
Linkage Map
1 2 3 4
Trait 2
Trait 1
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Mapping loci affecting quantitative traits (QTL) in
chickens
Distance along chromosome Gga 3 (cM)
Genes in the circled region appear to affect breast-meat yield
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High-throughput Genotyping
Illumina- BeadStation500G-BeadLab
~150,000 data points per week at UCDavis Genome Center
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Marker Assisted Breeding in Conifers
• Quantitative Trait Locus (QTL) Mapping • Association Mapping
Pinus taeda (loblolly pine)
Pseudotsuga menziesii (Douglas-fir)
Pinus elliottii(slash pine)
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Genomics & DNA Markers: Summary
• DNA markers can be used as fingerprints to distinguish individuals, and– cultivars, varieties, etc– increasingly used to protect intellectual property
(utility patents, PVP)
• Marker inheritance allows parentage to be verified, facilitating pedigree control
• DNA markers can be associated with phenotypic traits
• Once marker-trait associations have been established, marker data can augment phenotypic observations to accelerate breeding