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, limited by species boundaries, all genes/traits are mixed. ~45,000 genes~25,000 genes

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

GMO Introgression

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

Biolistics

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

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

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

Construct T-DNA selection genes virulence genes T-DNA (Transfer DNA) …gene of interest,  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 Transgenes must be expressed in order to function, Promoters control where, when and how much protein is produced. Foreign DNA is common (via nature) in most genomes,

Gene Structure chromosome (megabases) gene (kilobases)...ata cgt act atc... ||| ||| ||| |||...tat gca tga tag... protein coding...ttaggttctatc... ||||||||||||...aatccaagatag... promoter specific sequences.

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 “Suicide” Promoters, etc. Time Specific Expression

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 Bad Environmental Consequences Negative impacts on, select species, crops, ecosystems, etc. Works great Increase Carrying Capacity for Humans Human Population Growth Negative impacts on, select species, crops, ecosystems, etc.

Wednesday: Sugar and Genetics –PDF of paper available on WEB page, print pp Schedule Quiz on Below, Bring Paper for 2 pts.

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.

Quiz: 12.5 Midterm 2: mean = 69 (150, high score). GRADES

Assignment lecture 7 Do a Punnett Square or a Split Fork Diagram of, Parent 1: wild-type for Chromosomes 14, 21 x 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. SM1 4 : SM1 DNA sequence repeated 4 times, linked Signal: peptide sequence that sends protein to the midgut.

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.