The Science of Transgenics

The Sociological, Economic, and Ethical Impact

of Transgenic Organisms Workshop

Fargo, ND
February 21, 2003

Phil McClean
Department of Plant Science
North Dakota State University

NDSU
Extension
Transgenics are a Biotechnology Product
How about some definitions

Biotechnology - General Definition

The application of technology to improve
a biological organism

Biotechnology - Detailed Definition

The application of the technology to improve
the biological function of an organism
by adding genes from another organisms

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 What About the Term
Genetic Engineering?

Genetic engineering is the basic tool set of biotechnology

Genetic engineering involves:

 Isolating genes
 Modifying genes so they function better
 Preparing genes to be inserted into a new species
 Developing transgenes

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 What is a transgenic?

Concept Based on the Term Transgene

Transgene – the genetically engineered gene added to a species

Ex. – modified EPSP synthase gene (encodes a protein

that functions even when plant is treated with Roundup)

Transgenic – an organism containing a transgene

introduced by technological (not breeding) methods

Ex. – Roundup Ready Crops

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 Why are transgenics important?

We can develop organisms that express a “novel” trait

not normally found in the species

Extended shelf-life tomato (Flavr-Savr)

Herbicide resistant soybean (Roundup Ready)

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 Agriculture Transgenics On the Market

Insect resistant cotton – Bt toxin kills the

cotton boll worm
• transgene = Bt protein
Source: USDA

Insect resistant corn – Bt toxin kills the

European corn borer
• transgene = Bt protein
Normal Transgenic

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 Herbicide resistant crops
Now: soybean, corn, canola
Coming: sugarbeet, lettuce, strawberry
alfalfa, potato, wheat (2005?)
• transgene = modified EPSP synthase or
phosphinothricin-N-acetyltransferase
Source: Monsanto

Virus resistance - papaya resistant to

papaya ringspot virus
• transgene = virus coat protein

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 Biotech chymosin; the enzyme used
to curdle milk products
Source: Chr. Hansen
• transgene = genetically engineered enzyme

bST; bovin somatotropin; used to increase

milk production
• transgene = genetically engineered enzyme

Source: Rent Mother Nature

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Some Ag Biotech Products Are Discontinued
Why???

Poor Quality
• Flavr Savr tomatoes (Calgene)

Negative Consumer Response

• Tomato paste (Zeneca)

Negative Corporate Response

• New Leaf (Monsanto)

Universal Negative Publicity

• Star Link corn (Aventis)

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 Next Generation of Ag Biotech Products

Golden Rice – increased Vitamin A content

(but not without controversy)
transgene = three pathway enzymes

Sun flower – white mold resistance

transgene = oxalate oxidase from wheat
Source: Minnesota
Microscopy Society

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 Turfgrass – herbicide resistance;
slower growing (= reduced mowing)

Bio Steel – spider silk expressed in goats; used to

make soft-body bullet proof vests (Nexia)

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Biotechnology is Not Just on the Farm

Disease Treatment

Diagnostics
 
Environmental Cleanup
 
Human Applications

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 Human Applications

• Pharmaceutical products
New solutions to old problems
 
• Disease diagnosis
Determine what disease you have or may get 

• Gene therapy
Correcting disease by introducing a corrective gene

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 Biotechnology and Health
Product Use

Insulin Diabetes

Interferon Cancer

Interleukin Cancer

Human growth hormone Dwarfism

Neuroactive proteins Pain

The genes for these proteins are:

• Cloned
• Inserted into bacteria

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• Product isolated using biofermentation
Extension
Environmental Applications

Bioremediation - cleanup contaminated

sites; uses microbes designed to degrade
the pollution

Indicator bacteria – contamination can be

detected in the environment

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Future Health-related Biotech Products

Vaccines – herpes, hepatitis C, AIDS, malaria

Tooth decay – engineered Streptococcus mutans,

the bacteria that destroys enamel

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 Edible Vaccines
Transgenic Plants Serving Human Health Needs

• Works like any vaccine

• A transgenic plant with a pathogen protein gene is developed
• Potato, banana, and tomato are targets
• Humans eat the plant
• The body produces antibodies against pathogen protein
• Humans are “immunized” against the pathogen
• Examples:
Diarrhea
Hepatitis B
Measles

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 A Popular Term We Need To Know

GMOs - Genetically modified organisms

• GMO - an organism that expresses traits that result

from the introduction of foreign DNA

• Originally a term equivalent to transgenic organism

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 The GMO Ruse

 Some claim any improved biological product is a GMO

 They feel this will

• ease the publics fear
• pave the way for product acceptance

 For example, some call plant varieties biotechnology products

 This is a false claim

NDSU
Extension
 Let’s Be Up Front

• Biotechnology adds traits not available in the species

 Soybean does not have a gene to breakdown Roundup

 The gene comes from bacteria

• Breeding  Biotechnology

 Breeding only exchanges genes found in the species

 Breeding can transfer the transgene to other breeding materials
 BUT this does not make it a biotechnology procedure

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Extension
 Important Plant Improvement Methods

• Breeding
Crossing two individuals from the same species;
produces a new, improved variety;
Source: USDA not a biotechnology procedure

• Transformation
Adding a gene from another species; the
essential biotechnology procedure to produce
transgenics
Source: USDA

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 Interspecific Cross
Wheat Rye

Triticale
New species, but
NOT biotechnology
products

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 Mutagenesis

A useful procedure to produce a new trait

But the normal gene is modified

A transgene is not involved

 The product of mutagenesis is not a GMO

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Mutagenesis Changes the DNA Sequence

Mutagenesis
Treatment
Susceptible
Normal
Gene
ATTCGA

Resistant
Mutant
Gene
ATTGGA

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 BASF Clearfield Products

Herbicide resistance
•imidazolinones
Mutant AHAS enzyme
•developed by mutagenesis
Crops
• Canola
• Corn
• Rice
• Sunflower
• Wheat
A Major Marketing Advantage
but lost when stacked with a transgene

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Extension
 The Roundup Ready Story

• Glyphosate is a broad-spectrum herbicide

• Active ingredient in Roundup herbicide
• Kills all plants it come in contact with
• Inhibits a key enzyme (EPSP synthase) in an amino acid pathway

• Plants die because they lack the key amino acids

• A resistant EPSP synthase gene allows crops

to survive spraying

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Extension
 Roundup Sensitive Plants
Shikimic acid + Phosphoenol pyruvate
+ Glyphosate

X
Plant
EPSP synthase

Without amino acids,

X
3-Enolpyruvyl shikimic acid-5-phosphate
(EPSP)
plant dies

X
X
Aromatic
amino acids

NDSU
Extension
 Roundup Resistant Plants
Shikimic acid + Phosphoenol pyruvate
+ Glyphosate
RoundUp has no effect;
Bacterial enzyme is resistant to herbicide
EPSP synthase

3-enolpyruvyl shikimic acid-5-phosphate

(EPSP)
With amino acids,
plant lives

Aromatic
amino acids

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 The Golden Rice Story

• Vitamin A deficiency is a major health problem

• Causes blindness
• Influences severity of diarrhea, measles

• >100 million children suffer from the problem

• For many countries, the infrastructure doesn’t exist

to deliver vitamin pills

• Improved vitamin A content in widely consumed crops

an attractive alternative

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 -Carotene Pathway Problem in Plants
IPP

Geranylgeranyl diphosphate

Phytoene synthase

Phytoene
Problem: Phytoene desaturase
Rice lacks
these enzymes ξ-carotene desaturase

Lycopene
Lycopene-beta-cyclase
Normal
 -carotene

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Vitamin A
“Deficient” (vitamin A precursor)
Rice Extension
 The Golden Rice Solution
-Carotene Pathway Genes Added

IPP

Geranylgeranyl diphosphate

Daffodil gene Phytoene synthase

Phytoene
Vitamin A
Phytoene desaturase
Pathway Single bacterial gene;
is complete performs both functions
and functional ξ-carotene desaturase

Lycopene
Daffodil gene Lycopene-beta-cyclase

 -carotene

NDSU
Golden
Rice (vitamin A precursor)
Extension
 Introducing the Gene
or
Developing Transgenics

Steps

1. Create transformation cassette

2. Introduce and select for transformants

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 Transformation Cassettes

Contains

1. Gene of interest
• The coding region and its controlling elements

2. Selectable marker
• Distinguishes transformed/untransformed plants

3. Insertion sequences
• Aids Agrobacterium insertion

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Extension
 Transformation Steps

Prepare tissue for transformation

• Tissue must be capable of developing into normal plants
• Leaf, germinating seed, immature embryos

Introduce DNA
• Agrobacterium or gene gun

Culture plant tissue

• Develop shoots
• Root the shoots

Field test the plants

• Multiple sites, multiple years

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 Delivering the Gene
to the Plant
• Transformation cassettes are developed in the lab

• They are then introduced into a plant

• Two major delivery methods

• Agrobacterium

Tissue culture
• Gene Gun required to generate
transgenic plants

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The Lab Steps

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The Next Test Is The Field
Herbicide Resistance

Non-transgenics

Transgenics

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Final Test of the Transgenic
Consumer Acceptance

RoundUp Ready Corn

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Before After
Extension