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, transgenic organisms, other approaches, targeted mutagenesis, gene introgression, ? Old Science Humans (~30,000 years) Humans (~30 years) 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 technology is not essential, ~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.

Wild tomato Genome Era Traditional Breeding 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? insert into plant …via biolistics ...uses tools of molecular genetics, - i.e. applied bacteria and virus genetics. insert into plant …via biolistics - or - Agrobacterium tumefaceins

Biolistics

Agrobacterium tumefaciens Kalanchoe Stem w/ infection. Agrobacterium tumefaciens Natural soil bacterium that infects plants, hosts: 160 Genera, families: > 60, effect; poor growth, low yield.

Out: Ti genes, opine genes, Ti-Plasmid Transfer-DNA Plant Cells Nature Ti: tumor inducing Plasmid: extrachromosomal DNA evolved for genetic transfer. Hormone genes Opines genes Agrobacterium Lab Out: Ti genes, opine genes, In: DNA of choice. T-DNA Any Gene Selectable Markers, etc

Plant chromosome with T-DNA insert. T-DNA (Transfer DNA) …with gene of interest, b-carotene, herbicide resistance, etc.. Construct T-DNA transform, select for agro with T-DNA Agrobacterium infect plant, select for plants with T-DNA Plant chromosome with T-DNA insert.

Virulence genes: facilitate Agro infection, T-DNA transfer, T-DNA (Transfer DNA) selection genes …gene of interest, b-carotene, herbicide resistance, etc.. virulence genes Construct T-DNA 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... ||| ||| ||| ||| ...tat gca tga tag... ...ttaggttctatc... |||||||||||| ...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, 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, soil ecosystems, 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, select species, crops, ecosystems, etc. Negative impacts on, select species, crops, ecosystems, etc.

Transgenic Construct pBacR: piggyback vector, transposon derived 3xP3-EGFP-S40: Green fluorescent protein, eye specific promoter Signal: peptide sequence that sends protein to the midgut. SM14: SM1 DNA sequence repeated 4 times, linked AgCP promoter: mosquito promoter, activated by blood feast. pBacR: piggyback vector, transposon derived