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Understanding Biotechnology

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Title: Understanding Biotechnology


1
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

2
Outreach website
http//wwwdata.forestry.oregonstate.edu/orb/
3
Educational activities Food for Thought Lecture
Series / 2005-2008
  • Streaming video - OPAN/OPB usage

4
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

5
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.

6
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

7
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
8
Rapid rise of GE crops in developed and
developing world
http//www.isaaa.org
9
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

10
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

11
  • 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
12
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
13
Recombinant DNA modification of native plant genes
14
  • How are GE plants produced?

Step 1Getting whole plants back from cultured
cells cloning
15
  • Differentiation of new plant organs from single
    cells

First step is de-differentiation into callus
after treatment with the plant hormone auxin
Leaf-discs
16
  • Shoots, roots, or embryos produced from callus
    cells using plant hormones

17
  • Step 2

Getting DNA into plant cells
Main methods - Agrobacterium tumefaciens -
Biolistics gene gun
18
Agrobacterium is a natural plant genetic engineer
19
  • Agrobacterium gene insertion

Gene of interest
Engineered plant cell
Agrobacterium tumefaciens
20
  • 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)
21
  • Hormones in plant tissue culture
  • stimulate division from plant cells

22
  • 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)

23
Transformation of bentgrass(Wang and Ge 2006)
24
Glyphosate-tolerant FescueConventionally-bred
Patented Varieties
25
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

26
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27
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

28
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29
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

30
Strong and well funded political and legal
resistance
31
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

32
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

33
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

34
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

35
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

36
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

37
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

38
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

39
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)

40
Genomes, genes, and DNA
Genes are located on packaging platforms called
chromosomes
41
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42
A DNA fingerprint is fixed throughout an
individuals life
Age
43
DNA Fingerprints to Verify Identities 22 Paired
Samples Collected at Different Times
MCW-305
MCW-184
MCW-087
44
Pedigree errors non-parental marker types
Progeny
S D
45
Genetic Map Perennial Ryegrass
Gill et al. 2006
46
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
47
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48
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)
49
High-throughput Genotyping
Illumina- BeadStation500G-BeadLab
150,000 data points per week at UCDavis Genome
Center
50
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51
Marker Assisted Breeding in Conifers
  • Quantitative Trait Locus (QTL) Mapping
  • Association Mapping

52
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
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