Quinoa Mother Grain for Mother Earth

1. QuinoaMother Grain for Mother Earth

2. Quinoa and Downy Mildew • Quinoa is a highly nutritious and understated pseudo grain • Worshiped by the Incas as mother grain • 90 - 99% of Quinoa crops are ruined by downy mildew

3. Science and Technology Report Geneticist: Pseudo PhD. Catherine M. G. Murray

4. Science and Technology Report • Altered Plant: Quinoa (Chenopodium quinoa) • Goal: Resistance to downy mildew • Source of Gene: Sunflower (Helianthus annuus L.) • Gene PLH1

5. Step One – Isolating Gene PLH1 • Isolate the DNA of Helianthus annuus • Obtain a sample of sunflower plant tissue • Ground tissue in a blender, put in a tube • Add a lysis solution • Solution contains a detergent and proteinase K • Place tube in warm water bath

6. Step One – Isolating Gene PLH1 • Add a concentrate salt solution • Place tube in centrifuge • Use a micropipette to remove the liquid and put in another tube • Add isopropyl alcohol and gently mix. • Place the tube in a centrifuge • Use a micropipette to remove the liquid at the top of the tube • Result: extracted and isolated DNA

7. Step One (Cont) • Isolate the Gene PLH1 using restriction endonucleases • Restriction endonucleases - enzymes that splice the DNA at certain points where it recognizes specific sequences • Creates DNA fragments

8. Step Two – Cloning the Gene • This is done through the Polymerase Chain Reaction (PCR) video

9. Step Three – Designing Gene • Transcription – the synthesis of RNA from a DNA template • Three parts of a gene • Promoter • Coding Region • Termination Sequence • RNA polymerase – enzyme that “reads” the DNA code and builds the mRNA strand

10. Step Three – Designing Gene (Cont) • Transcription Factor – factors that are needed to produce the mRNA strand or “transcript” • Process: • RNA polymerase binds to DNA strand • Double helix unwinds • RNA polymerase moves along the DNA strand (3’ ---> 5’ direction)

11. Step Three – Designing Gene (Cont) • Ribonucleotides bond to the DNA strand • Result: strand of mRNA • Strand of ribonucleotides begins to break away • DNA strands rewind into double helix • RNA polymerase reaches the termination sequence • RNA polymerase and the mRNA strand released from the DNA strand • mRNA strand -----> cytoplasm

12. Step Four – Inserting PLH1 • “Gene Gun Method” • Paint copies of the gene PLH1 onto microscopic metal (tungsten) particles • Load onto a gene gun

13. Step Four (Cont) • Fire at cells of Quinoa – “callus” • Metal particles penetrate cells • Cellular Fluid flushes DNA off particles • DNA -----> nucleus ------> integrated into the genome

14. Step Five – Backcross Breeding • Transgenic Quinoa plant is inbred • Sunflower plant is inbred • Transgenic inbred line x elite inbred line ----> F1 generation • 50% of the transgenic line’s alleles and 50% of the elite line’s alleles • F1 generation x elite inbred line ----> BC1 generation

15. Step Five (Cont) • 75% elite line’s alleles and 25% transgenic line’s alleles • ELISA TEST • Crossing continues until BC5 • Percentage of elite alleles is about 99% • Last step: self-cross the new transgenic elite line

16. Product • A new line of Quinoa plants in which the downy mildew-resistant gene has been incorporated

17. Consumer Appeal Analysis Ethisist: Rebecca A. Crawford

18. Public Ethics Concerns According to the Food and Agricultural Organization of the UN, the most important ethical values of food are: The value of food The value of enhanced well-being The value of human health The value of natural resources The value of nature

19. The Value of Food • Every year 15 million children die of hunger • Good for countries where nutrition is scarce

20. The Value of Enhanced Well-Being and Human Health • Help poor farmers in south America and Asia • Boost economy • Increase income • Extremely healthy: • Good source of magnesium, iron, copper and phosphorus

21. Common Public Concerns • Use of “material from organisms that have never been part of the human food supply” • Toxins • Allergic Reactions • Decreased Nutritional Value

22. How We Address Them • Use of “material… [not]…part of the human food supply” • We use sunflower genes • Both are plants • Sunflowers – commonly consumed

23. How We Address Them (Cont) • Toxins • The pesticide for downy mildew (Mancozeb) now used on quinoa can cause: • Reproductive and developmental toxicity • Neurotoxicity • Acute toxicity • Cancer • Inserting PLH1 would eliminate need for toxins

24. How We Address Them (Cont) • Allergic Reactions • 90 - 99% more quinoa would help more than harm • Sunflower allergies are extremely uncommon • Quinoa helps people with allergies • 1 in 133 Americans - Celiac disease • Many non-related gluten allergies • Appropriately label the GM quinoa

25. How We Address Them (Cont) • Decreased Nutritional Value • We are eliminated a health risk • Besides Quinoa’s nutritional value, sunflowers are good sources of: • A Rich Source Of Vitamin E • Dietary Fiber • Proteins • Carbohydrates

26. Environmental Impact Statement Environmentalist: Jenna n. bates

27. Environmental Impact • Not completely known • Will take several years or decades • Vary from plant to plant • Only real world information can show the true effects • Monitor plants closely

28. Environmental Impact (Cont) • The downy mildew could become resistant • Gene transfer to wild plants • Low risk because Quinoa has few wild relatives in the Himalayas • Native to Peru • No hybrid between GM and wild plants has survived in the wild • Gene transfer to weeds

29. Environmental Impact (Cont) • Unlikely to improve strength of weed • The natural pesticides it makes could kill animals • The risk of this is very low • Use of pesticides would actually pose a more evident risk to the surrounding ecosystem. • Reduce pesticide use

30. Environmental Impact (Cont) • “GM crops have enabled U.S. farmers to make annual reductions in pesticide applications of 46 million pounds since 1996.” • Mancozeb kills animals • Reduction of fosil fuel use • Less soil erosion • Unknown effects on soil and the flow of nutrients

31. Marketing Plan Agricultural Economist: Alexandra M. C. Stambaugh

32. SUNFLOWER ENHANCED • Genetically altered plants are closer to the hybrid plants than the actual plants • What is the purpose of the modification? • 99% of the Quinoa plant crop is eliminated by Downy Mildew

33. Who is our market? • Marketable in countries • Japan- macrobiotic diet (25-30% of diet cereals and grains).  • Other Eastern Asian countries such as India and China  • “Here in Henan's rural Gushi County…” • Market in Developed Countries—market similarly to wholegrain, with protein and fiber

34. Problem Addressed and Solution • Fighting malnutrition on a low budget: • Economic price: cooked 222 calories for cup (229g) cooked can be priced around $3 for a .5 kg bag.  • Important for a Vegetarian diet—8 + grams of protein per cup • Contains all 9 essential amino acids, 5 grams of fiber (more than prunes, whole wheat, rye)

35. Problem Addressed and Solution (Cont) • No saturated fats, no transfats, extremely low sodium, no sugars • Can be grown in high elevations—specifically 10,000 ft • Easily grown in Himalayas—stimulates economic growth in Eastern Asian countries

36. Problem Addressed and Solution (Cont) • Only issue is water logging, which is easy to revise • Easy to grow- mature seeds germinate once in contact with water

37. How is it going to be marketed? • Quinoa must be rinsed before packaging—Saponin – natural pesticide • Dried and then packaged in bags or boxes • Versatility of use • As is, Flour, Pasta, many other grain products • Easy storage: cool, dry place, Storage calls for easy transport

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39. Bibliography "The Polymerase Chain Reaction." Sumanas, Inc.. 2004. Sumanas. 12 Jan 2009 http://www.sumanasinc.com/webcontent/animations/content/pcr.html. "Restriction Enzymes." UCDavis. 12 Jan 2009 http://ppge.ucdavis.edu/acrobat/restriction_enzymes.pdf. "Methods for Genetically Engineering a Plant." 2000. U.S. Drug and Food Administration. 12 Jan 2009 <http://www.fda.gov/fdac/features/2000/biochart.html>;. "Polymerase Chain Reaction." Gene Almanac. Cold Spring Harbor Laboratory. 12 Jan 2009 <http://www.dnalc.org/ddnalc/resources/pcr.html>;. Hain, Patty. "Transcription Detail." 2000. University of Nebraska. 12 Jan 2009 <http://www-class.unl.edu/biochem/gp2/m_biology/animation/gene/gene_a2.html>;. "DNA Isolation Methods." 2009. enotes.com. 12 Jan 2009 <http://www.enotes.com/forensic-science/dna-isolation-methods>;. "Backcross Breeding." Library of Crop Technology Lesson Modules. 12 Jan 2009 <http://croptechnology.unl.edu/viewLesson.cgi?min=1&max=9&topic_order=3&LessonID=959009357>;. Woods, Georgia. "Preparation of Gene Gun Bullets and Biolistic Transfection of Neurons in Slice Culture." Journal of Visualized Experiments 1212 Feb 2008 12 Jan 2009 <http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2582847>;. "Concept 6: The Transcription Process." The Biology Place. 2008. Pearson Education, Inc. 12 Jan 2009 <http://www.phschool.com/science/biology_place/biocoach/transcription/tcproc.html>;.

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