Understanding Biotechnology

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Understanding Biotechnology

• Steve Strauss, Professor, OSU
• Forest Science, Genetics, Molecular and Cellular
  Biology
• Director, Outreach in Biotechnology
• http//wwwdata.forestry.oregonstate.edu/orb/
• Steve.Strauss_at_OregonState.Edu

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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
• Its 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
• Thats why its called genetic engineering, though
  we are modifying, not building, a new organism

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• Assembling a gene

Protein

• Provides stability to messenger RNA, and
• Guides processing into protein

• Controls level of expression, and
• Where and when expressed

Can mix and match parts can change sequences to
improve properties
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Examples of promoter gene combinations
produced via recombinant DNA methods
Promoter (controls expression)
Gene (encodes protein)
Phenolic pathway enzyme (bacteria)
35S-CAMV (plant virus)
Herbicide tolerant
RNA degrading enzyme (bacteria)
Pollen sac (tobacco)
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

First step is de-differentiation into callus
after treatment with the plant hormone auxin
Leaf-discs
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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
Engineered plant cell
Agrobacterium tumefaciens
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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 culture
• stimulate division from plant cells

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• Antibiotics in plant tissue culture
• limit growth to engineered cells
• Other 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
turfgrassesWang 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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Varied public approval

• 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

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Rutgers survey data - USA (2005)http//www.foodpo
licyinstitute.org/resultpub.php
http//www.foodpolicyinstitute.org/docs/reports/N
ationalStudy2003.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 Markers

• David Harry
• Department of Forest Science
• Assoc. Director, Outreach in Biotechnology
• http//wwwdata.forestry.oregonstate.edu/orb/
• david.harry_at_oregonstate.edu

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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 100s to 1000s 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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Genomes, genes, and DNA
Genes are located on packaging platforms called
chromosomes
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A DNA fingerprint is fixed throughout an
individuals life
Age
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DNA Fingerprints to Verify Identities 22 Paired
Samples Collected at Different Times
MCW-305
MCW-184
MCW-087
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Pedigree errors non-parental marker types
Progeny
S D
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Genetic Map Perennial Ryegrass
Gill et al. 2006
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How might genetic markers accelerate breeding?
X
X
X
X
Then, evaluate genetic makeup early to select
young birds
First, associate performance and genetic makeup
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Mapping loci affecting quantitative traits (QTL)
in chickens
Genes in the circled region appear to affect
breast-meat yield
Distance along chromosome Gga 3 (cM)
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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

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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