Genetically Modified Plants

Biotechnology: underlying science

Potential Risks vs.(Potential) Benefits

Assigned Reading: Chapter 10.5

 Genetically Modified Organisms
Types of GMOs?
- artificial selection and
traditional breeding,
Old Science
- transgenic organisms,

- other approaches, Humans (~30,000 years)

- targeted mutagenesis,
Humans (~30 years)
- gene introgression, Bacteria (eons)
- ?
Humans (~15 years)
Bacteria (eons)
 Desirable Agronomic Traits
(traditional or modern)

• Increased yields, more nutritious, quality, etc.,

• More resistant to pestilence, weeds, water and

nutrient deprivations,

• Ability to withstand marginal growth conditions,

– and thrive in new environmental ranges,

• Profit.
 Traditional Breeding

~25,000 genes ~45,000 genes

• technology is not essential,

• limited by species boundaries,
• all genes/traits are mixed.
 Introgression
…incorporation of genes of one
genome into the genome of another
cultivar,
– standard breeding techniques are
laborious (if possible at all),

– genomics and related sciences greatly

accelerates standard breeding techniques.
 Genome Era Traditional Breeding

Wild tomato Cultivar w/ 1 wild gene replacement

Genetic Bottlenecks and Seed Preservation
Introgression GMO
 Transgenic Plants

• based on DNA technology,

• single genes/traits can be transferred,
• species boundaries are not limiting.
 How are GMOs generated?

...uses tools of
molecular genetics,
- i.e. applied
bacteria and
virus genetics.

insert into plant

…via biolistics - or - Agrobacterium tumefaceins

Biolistics
 Kalanchoe Stem
w/ infection.

Agrobacterium tumefaciens

Natural soil bacterium

that infects plants,
hosts: 160 Genera,
families: > 60,
effect; poor growth,
low yield.
 Ti-Plasmid Transfer-DNA Plant Cells
Nature
Ti: tumor inducing

Plasmid:
extrachromosomal
DNA evolved for Hormone Opines
genes genes
genetic transfer. Agrobacterium

Lab
T-DNA

Out: Ti genes, opine genes,

Any Gene
In: DNA of choice. Selectable Markers, etc
T-DNA (Transfer DNA)

…with gene of interest,

carotene,
Construct T-DNA - herbicide resistance, etc..

transform, select for agro with T-DNA

Agrobacterium

infect plant, select for plants with T-DNA

Plant chromosome with T-DNA insert.

T-DNA (Transfer DNA)
selection genes

…gene of interest,
Construct T-DNA virulence
genes carotene,
- herbicide resistance, etc..

Virulence genes: facilitate Agro infection, T-DNA transfer,

• not usually transferred in commercial applications,

Selection genes (2+): used to identify transgenics,

• usually antibiotic or herbicide resistance, etc. (i.e. only the
organisms with the T-DNA live in a selection experiment),

Gene of interest: protein coding region, plus a “promoter”.

Promoters Control Expression

Foreign DNA is common (via nature) in most genomes,

Transgenes must be expressed in order to function,

Promoters control where, when and how much protein is produced.
 Gene Structure

chromosome
(megabases)

gene (kilobases)

...ata cgt act atc...

...ttaggttctatc...
||| ||| ||| |||
||||||||||||
...tat gca tga tag...
...aatccaagatag...

promoter specific sequences. protein coding

 Promoter Specifies Expression

General Promoter: all tissues, all the time.

Vegetative Promoter: no flower, no fruit expression.

Root Promoter: only root expression.

Expression = Protein Production

Protein and
protein functions
only present in
tissue with
active promoter.

Tissue Specific Expression

Time Specific Expression

“Suicide” Promoters, etc.
Brief History of Transgenic
Organisms
• Transgenic E. coli,

– not demonstratively dangerous,

– demonstratively beneficial (probably).

• Transgenic virus,

– not demonstratively dangerous,

– demonstratively beneficial (probably).

• Transgenic plants,

– demonstratively dangerous? (not yet),

– demonstratively beneficial (?).
 Potential Risks
• Risk of invasion.
• Direct nontarget Effects

• Indirect nontarget Effects.

• New Viral Diseases.

• Variability and Unexpected Results.

 Potential Risks
(risk of invasion)
• 50,000 invaders in USA the old
fashioned ways,

– self-sustaining cultivars,
• low anticipated risk,

– hybridization with (native)

neighbors,
• transgene introgression,
• introgression of domestic cultivar genes
with natives has occurred, resulting in
negative impacts on native species,

– time lags.
 Direct
(nontarget)

• Risk to non-target species,

– pollinators,
– passers-by,

• soil ecosystems,

– decomposition rates,
– carbon cycle,
– nitrogen cycle.
 Indirect
(nontarget)

• kill weeds = kill species that

live “on” or eat the weeds,

• bioaccumulation,
– nontarget species eat plants,
store toxins,
– those species are eaten,
amassing the toxin,
– on up the food chain.
Bee on Red Clover.
 New Viral Diseases

• virus resistant plants promote

virulent strains,

– mutations,
– recombination,

• heteroencapsulation,

– virus move genes from one

organism to another,
– not presently a risk, but a
potential risk.
Variability and Unexpected Results
• time scale,

• numbers,

• environmental and cultivar

differences,

• application, culture and

consistency.
 Other Issues
• Economic hegemony of GMP seed producing countries,
companies,

• Cultural shifts in farming due to the introduction of GMOs,

• Potential allergies to genetically modified crops,

• The preservation of natural genetic crop-lines,

• The lack of an adequate risk assessment methodology to

quantify unintended ecological consequences.
The Precautionary Principle
 Biotechnology in General
Scenario 1 Scenario 2

Works great
Bad Environmental Consequences
Increase Carrying Capacity for Humans

Human Population Growth

Negative impacts on,
Negative impacts on,
• select species,
• crops,
• select species,
• ecosystems,
• crops,
• etc.
• ecosystems,
• etc.
 Schedule
Quiz on Below, Bring Paper for 2 pts.

• Wednesday: Sugar and Genetics

– PDF of paper available on WEB page,

• print pp. 7 - 15.

 Rest of Quarter
assignments
• June 1st: Chapter 8 and 9 reading assignments for
background.

• June 4th: Human and Chimps Paper.

• June 6th: Pheomone Paper.

• On Final: PCR, Northerns, Southerns,

Sequencing, and other relavant techniques.
GRADES Quiz: 12.5
Midterm 2:
mean = 69 (150, high score).
 Assignment
lecture 7

• Do a Punnett Square or a Split Fork Diagram

of,

Parent 1: wild-type for Chromosomes 14, 21

Parent 2: heterozygous for 14q;21q translocation.

Hint

gametes
 Transgenic Construct

pBacR: piggyback vector, transposon derived

3xP3-EGFP-S40: Green fluorescent protein, eye specific promoter

AgCP promoter: mosquito promoter, activated by blood feast.

Signal: peptide sequence that sends protein to the midgut.

SM14: SM1 DNA sequence repeated 4 times, linked

 What You Can Do
1. Pick one question on the exam that you missed,
answer completely for full credit,
• must be clearly and completely answered,
• Return Wednesday, 12 noon, stapled to exam.

2. Do better on the final, and get 1/2 the difference

between Exam 2 and the Final,
• Example: 100 on E#2, 150 on Final, then add 25 pts.