Understanding Biotechnology

Steve Strauss, Professor, OSU

Forest Science, Genetics, Molecular and Cellular Biology

Director, Outreach in Biotechnologyhttp://wwwdata.forestry.oregonstate.edu/orb/

Steve.Strauss@OregonState.Edu

Outreach website

http://wwwdata.forestry.oregonstate.edu/orb/

Educational activities Food for Thought Lecture Series / 2005-2008

Streaming video - OPAN/OPB usage

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

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.

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

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

Rapid rise of GE crops in developed and developing world

http://www.isaaa.org

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

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

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

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

Recombinant DNA modification of native plant genes

How are GE plants produced?

Step 1Getting whole plants back from cultured

cells = cloning

Differentiation of new plant organs from single cells

Leaf-discs

First step is de-differentiation into “callus” after treatment with the plant hormone auxin

Shoots, roots, or embryos produced from callus cells using plant hormones

Step 2

Getting DNA into plant cells

Main methods- Agrobacterium tumefaciens- Biolistics [gene gun]

Agrobacterium is a natural plant genetic engineer

Agrobacterium gene insertion

Gene of interest

Agrobacterium tumefaciens

Engineeredplant cell

T-DNA

Ti Plasmid

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)

Hormones in plant tissue culturestimulate division from plant cells

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)

Transformation of bentgrass(Wang and Ge 2006)

Glyphosate-tolerant FescueConventionally-bred Patented Varieties

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

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

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

Strong and well funded political and legal resistance

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

• 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

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

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

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

Understanding Biotechnology

Part 2:

Genomics and DNA MarkersDavid Harry

Department of Forest ScienceAssoc. Director, Outreach in Biotechnology

http://wwwdata.forestry.oregonstate.edu/orb/

david.harry@oregonstate.edu

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

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

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)

Genes are located on packaging platforms called

chromosomes

Genomes, genes, and DNA

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

T

C

CA

C

G T

T

G<>C

G<>T

Marker “1”

Marker “2”

AgeAge

A DNA fingerprint is fixed throughout an individual’s

life

MCW-305

MCW-184

MCW-087

DNA Fingerprints to Verify Identities 22 Paired Samples Collected at Different Times

S DProgeny

Pedigree errors: “non-parental” marker types

Genetic Map: Perennial Ryegrass

Gill et al. 2006

X XThen, evaluate genetic makeup early to select

young birdsFirst, associate performance and

genetic makeup

X X

How might genetic markers accelerate breeding?

a

b

c

d

e

f

g

h

i

j

k

l

m

Hypothetical genes (QTLs) affecting economic traits

Linkage Map

1 2 3 4

Trait 2

Trait 1

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

High-throughput Genotyping

Illumina- BeadStation500G-BeadLab

~150,000 data points per week at UCDavis Genome Center

Marker Assisted Breeding in Conifers

• Quantitative Trait Locus (QTL) Mapping • Association Mapping

Pinus taeda (loblolly pine)

Pseudotsuga menziesii (Douglas-fir)

Pinus elliottii(slash pine)

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