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BIO403: Biotechnology

Unit 5: Genetic Engineering of Plants   Plants have long been engineered for human use ever since the earliest civilizations established agricultural-based societies.  Most of the cultivated crops that we eat as food have already been artificially selected for thousands of years for certain characteristics.  With modern biotechnology, we have turned this largely trial-and-error process into a major science as we understand the functions of more and more genes and develop more sophisticated methods for introducing exotic genes into plants.  The new feature of the modern technology is the ability to produce transgenic plants.  For example, the addition of certain vitamins into rice plants has the potential to solve endemic malnutrition for millions of people in third-world countries.  This unit will also examine the safety and health issues surrounding genetically engineered plants.

Unit 5 Time Advisory
This unit should take you approximately 10 hours to complete.

☐    Subunit 5.1: 0.5 hour

☐    Subunit 5.2: 1.0 hour

☐    Subunit 5.3: 1.0 hour

☐    Subunit 5.4: 1.0 hour

☐    Subunit 5.5: 2.5 hours

☐    Subunit 5.6: 2.0 hours

☐    Subunit 5.7: 1.5 hours

☐    Subunit 5.8: 0.5 hour

Unit5 Learning Outcomes
Upon successful completion of this unit, the student will be able to:

  • Compare and contrast traditional plant breeding and transgenic technology.
  • Define the crucial characteristics of totipotent and multipotent cells.
  • Describe the Ti plasmid, the promoter sequences, and the reporter genes that are used in the production of transgenic plants.
  • Identify examples of genetically modified plants.
  • Discuss safety issues and ethical concerns regarding genetically modified plants.

5.1 Plant Breeding   5.1.1 Cross-Pollination   - Reading: National Center for Biotechnology Information’s Bookshelf: Sinauer Associates: S.F. Gilbert’s Developmental Biology, 6th edition: “Pollination” Link: National Center for Biotechnology Information’s Bookshelf:  Sinauer Associates: S.F. Gilbert’s Developmental Biology, 6th edition:  “Pollination” (HTML)
 
Instructions: Please study this excerpt from Giblert’s textbook in its entirety.
 
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5.1.2 Selective Breeding   - Reading: University of Notre Dame: Lauren Willoughby’s “Selective Breeding and Hybridization” Link: University of Notre Dame: Lauren Willoughby’s “Selective Breeding and Hybridization” (HTML)
 
Instructions: Please study this page.  Please note that humans have modified living organisms historically.
 
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5.1.3 Mutation Breeding   - Reading: National Center for Biotechnology Information’s Bookshelf: W.H. Freeman: Griffiths, Miller, Suzuki, et al.’s Introduction to Genetic Analysis, 7th edition: “Mutation Breeding” Link: National Center for Biotechnology Information’s Bookshelf:  W.H. Freeman: Griffiths, Miller, Suzuki, et al.’s Introduction to Genetic Analysis, 7th edition:  “Mutation Breeding” (HTML)
 
Instructions: Please study this section on mutation breeding from Griffiths, Miller, Suzuki, et al.’s textbook.
 
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5.2 Plant Cultures and Properties   5.2.1 Totipotency   - Reading: Dave’s Garden: LariAnn Garner’s “Totipotency—The Wonder of Stem Cells in Plants” Link: Dave’s Garden:  LariAnn Garner’s “Totipotency—The Wonder of Stem Cells in Plants” (HTML)
 
Instructions: Please study this page.
 
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5.2.2 Callus Culture   - Reading: University of Florida: Alba Myers’ “Somatic Embryogenesis Induction in Delonix Regia (Boger.) Raf (Royal Poinciana)” Link: University of Florida: Alba Myers’ “Somatic Embryogenesis Induction in Delonix Regia (Boger.) Raf (Royal Poinciana)” (HTML)
 
Instructions: Please read this page, and identify examples of callus culture.  Plant tissues are sterilized and placed on tissue culture medium, which induces “callus,” that is, undifferentiated cell formation.
 
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5.2.3 Suspension Culture   - Reading: The University of New South Wales’ Cell Biology Wiki: Dr. Mark Hill’s “Group 4 Project—Cell Culture” Link: The University of New South Wales’ Cell Biology Wiki: Dr. Mark Hill’s  “Group 4 Project—Cell Culture” (HTML)
 
Instructions: Please study the “Harvesting and Isolating Cells” section on this page.
 
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5.3 Insertion of Genes   5.3.1 Ti Plasmid   - Reading: National Center for Biotechnology Information’s Bookshelf: W.H. Freeman: Griffiths, Gelbart, Miller, et al.’s Modern Genetic Analysis: NCBI Bookshelf: Griffiths et al.’s “Recombinant DNA Technology in Eukaryotes” Link: National Center for Biotechnology Information’s Bookshelf:  W.H. Freeman: Griffiths, Gelbart, Miller, et al.’s Modern Genetic Analysis: NCBI Bookshelf: Griffiths et al.’s “Recombinant DNA Technology in Eukaryotes” (HTML)
 
Instructions: Please study “The Ti plasmid” section on this page.  The Ti plasmid is actually inserted by a bacteria species called Agrobacterium.  Thus, when we talk about the Ti plasmid, we are referring to the plasmid inside Agrobacterium. 
 
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5.3.2 Electroporation, Particle Bombardment, Microinjection   - Reading: National Center for Biotechnology Information’s Bookshelf: W.H. Freeman: Berg, et al.’s Biochemistry, 5th edition: “Manipulating the Genes of Eukaryotes” Link: National Center for Biotechnology Information’s Bookshelf:  W.H. Freeman: Berg, et al.’s Biochemistry, 5th edition: Manipulating the Genes of Eukaryotes” (HTML)
 
Instructions: Please read the third and fourth paragraphs (starting with “Foreign DNA can be introduced . . . “) in the “6.3.6 Tumor-Inducing Plasmids Can Be Used to Introduce New Genes into Plant Cells” section.
 
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  • Reading: Saint John’s University: Stephen G. Saupe’s “Introduction to the Protoplast Lab” Link: Saint John’s University:  Stephen G. Saupe’s “Introduction to the Protoplast Lab” (HTML)
     
    Instructions: Please study the “A Protoplast Primer” section on this page.
     
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  • Reading: National Center for Biotechnology Information’s Bookshelf: W.H. Freeman: Griffiths, Miller, Suzuki, et al.’s Introduction to Genetic Analysis, 7th edition: “Recombinant DNA Technology in Eukaryotes” Link: National Center for Biotechnology Information’s Bookshelf:  W.H. Freeman: Griffiths, Miller, Suzuki, et al.’s Introduction to Genetic Analysis, 7th edition:  “Recombinant DNA Technology in Eukaryotes” (HTML)
     
    Instructions: Please study the “Transgenic Eukaryotes” section on this page.
     
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5.4 Detection of DNA Insertion   5.4.1 Reporter Genes   - Reading: University of Central Florida: Dheeraj Verma and Henry Daniell’s “Chloroplast Vector Systems for Biotechnology Applications” Link: University of Central Florida: Dheeraj Verma and Henry Daniell’s “Chloroplast Vector Systems for Biotechnology Applications” (HTML or PDF)
 
Instructions: Please study the “Reporter Genes Used In Plastids” section on this page.  You can access the PDF from the top right corner of the page.
 
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  • Reading: Wellesley College’s “?-Glucuronidase (GUS) Activity Assays of Transformed Plants” Link: Wellesley College’s “β-Glucuronidase (GUS) Activity Assays of Transformed Plants” (HTML)
     
    Instructions: Please study this page.  Please note that β-glucuronidase is a reporter gene, which can be used to assess the succes of  transformation.
     
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  • Assessment: The Saylor Foundation's "BIO403 Unit 5.4 Assessment" Link: The Saylor Foundation's "BIO403 Unit 5.4 Assessment" (HTML)
     
    Instructions: You will find link to the "GM Plant Design" assessment on this page.  This is a multiple choice assessment with one correct answer.  Clicking on an answer will bring you to another page.  If your answer is correct, then it is acknowledged with a short explanation.  Please read the explanation carefully.  If you clicked on the wrong answer, then you will be directed to a tutorial page.  Please study the tutorial page carefully.  You will be prompted to return to the assessment and complete it again.  Please note that plastid modification is a more recent development in GM plant.design.

5.4.2 Cre-loxP System   - Reading: The Jackson Laboratory’s “Introduction to Cre-Lox Technology” Link: The Jackson Laboratory’s “Introduction to Cre-Lox Technology” (HTML)
 
Instructions: Please study this page. Note that Cre-Lox Technology is used both on animals and plants.
 
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5.5 Common Transgenic Properties   5.5.1 Herbicide Resistance   - Reading: International Service for the Acquisition of Agri-Biotech Applications: “Pocket K No. 10: Herbicide Tolerance Technology: Glyphosate and Glufosinate” Link:  International Service for the Acquisition of Agri-Biotech Applications:  “Pocket K No. 10: Herbicide Tolerance Technology: Glyphosate and Glufosinate” (HTML or PDF)
 
Instructions: Please study this page.  You can access the PDF in various languages from the left-hand side of the page.
 
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5.5.2 Insect Resistance   - Reading: John W. Kimball’s “Bacillus thuringiensis (Bt)” Link: John W. Kimball’s “Bacillus thuringiensis (Bt)” (HTML)
 
Instructions: Please study this page.
 
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  • Reading: Nature Education’s Scitable: Losey et al.’s “Transgenic Pollen Harms Monarch Larvae” Link: Nature Education’s Scitable: Losey et al.’s “Transgenic Pollen Harms Monarch Larvae” (PDF)
     
    Instructions: Please study this publication in its entirety.
     
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5.5.3 Stress Tolerance   - Reading: Virginia Tech: Information Systems for Biotechnology: Teresa Capell’s “Enhanced Drought Tolerance In Transgenic Rice” Link: Virginia Tech:  Information Systems for Biotechnology: Teresa Capell’s “Enhanced Drought Tolerance In Transgenic Rice” (HTML)
 
Instructions: Please study this page.  Author Theresa Chapel is from Department of Crop Genetics and Biotechnology, Schmallenberg, Germany.
 
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5.5.4 Crop Yield   - Reading: The NCAT Sustainable Agriculture Project’s “Transgenic Crops” Link: The NCAT Sustainable Agriculture Project’s “Transgenic Crops” (HTML)
 
Instructions: Please study the “Crop Yield, Costs, and Profitability” section on this page.
 
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  • Reading: Nature Education’s Scitable: Leslie A. Pray’s “Quantitative Genetics: Growing Transgenic Tomatoes” Link: Nature Education’s Scitable:  Leslie A. Pray’s “Quantitative Genetics: Growing Transgenic Tomatoes” (HTML)
     
    Instructions: Please study the entire article. Please note the similarities and differences between complex quantitative traits and single gene determined traits.
     
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5.5.5 Nutrition   - Reading: International Service for the Acquisition of Agri-Biotech Applications: “Biotechnology and Biofortification” Link:  International Service for the Acquisition of Agri-Biotech Applications: “ Biotechnology and Biofortification” (HTML or PDF)
 
Instructions: Please read the content of this page.  You can access the PDF from the left-hand side of the page.
 
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  • Reading: The American Society for Nutritional Sciences: The Journal of Nutrition: Bo Lönnerdal’s “Genetically Modified Plants for Improved Trace Element Nutrition” Link:  The American Society for Nutritional Sciences:  The Journal of Nutrition:  Bo Lönnerdal’s “Genetically Modified Plants for Improved Trace Element Nutrition” (HTML or PDF)
     
    Instructions: Please study the “Insertion of Genes for Novel Metal-Binding Proteins” section on this page. You can access the PDF form from the right side of the page.  Author Bo Lönnerdal works in the Department of Nutrition at the University of California.  This is a peer-reviewed publication.
     
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5.6 Functional Genomics of Plants   5.6.1 Metabolic Pathways   - Reading: Jim M. Dunwell’s “Transgenic Approaches to Crop Improvement” Link: Jim M. Dunwell’s “Transgenic Approaches to Crop Improvement” (HTML or PDF)
 
Instructions: Please study the content of this page. You can access the PDF form on the righthand side of the page.

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  • Reading: John W. Kimball’s Biology Pages: “Arabidopsis Thaliana: Another ‘Model Organism’” Links: John W. Kimball’s Biology Pages:  “Arabidopsis Thaliana: Another ‘Model Organism’” (HTML)
     
    Instructions: Please study this page.  Please note that A. thaliana is the most widely used model organism in research aiming to produce new genetically modified plants.
     
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5.6.2 Mutagenesis   - Reading: Oxford University Press’s Journal of Experimental Botony: Parry et al.’s “Mutation Discovery for Crop Improvement” Link: Oxford University Press’s Journal of Experimental Botony:  Parry et al.’s “Mutation Discovery for Crop Improvement” (HTML or PDF)
 
Instructions: Please study the “Introduction” and “Conclusions” sections on this page. You can access the PDF from the right side of the page.  This is a peer-reviewed publication.
 
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5.6.3 Gene Knockouts   - Reading: Arizona Genomics Institute, Arizona Genomics Computational Laboratory, and Stanford University: “The Maize Full Length cDNA Project” Link: Arizona Genomics Institute, Arizona Genomics Computational Laboratory, and Stanford University: “The Maize Full Length cDNA Project” (HTML)
 
Instructions: Please study the “Disrupt Gene Regulation: ‘Knockdown’ Technology” section on this page.
 
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5.7 Genetically Modified Plants   5.7.1 Tomatoes   - Reading: National Education’s Scitable: Leslie A. Pray’s “Quantitative Genetics: Growing Transgenic Tomatoes” Link: National Education’s Scitable:  Leslie A. Pray’s “Quantitative Genetics: Growing Transgenic Tomatoes” (HTML)
 
Instructions: Please study the content of this page.  Longer shelf life, size, and taste are some of the traits that are targeted in transgenic tomatoes.  Please note the broad range of traits, which transgenic technology attempts to modify.
 
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5.7.2 Soybeans   - Reading: National Center for Biotechnology Information: The Yonsei University’s College of Medicine: Yonsei Medical Journal: Sang-Ha Kim, et al.’s “Evaluating the Allergic Risk of Genetically Modified Soybean” Link: National Center for Biotechnology Information: The Yonsei University’s College of Medicine: Yonsei Medical Journal:  Sang-Ha Kim, et al.’s “Evaluating the Allergic Risk of Genetically Modified Soybean” (HTML or PDF)
 
Instructions: Please study the “Introduction” and “Discussion” sections on this page. You can access the PDF form from the top left corner of the page.  This is a peer-reviewed publication.  Please note that possible allergic reaction to transgenic soybean is discussed.
 
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5.7.3 Corn   - Reading: PLoS Biology: Virginia Gewin's “Genetically Modified Corn— Environmental Benefits and Risks” Link: PLoS Biology: Virginia Gewin's “Genetically Modified Corn— Environmental Benefits and Risks” (HTML or PDF)
 
Instructions: Please study this page. You can download this material in PDF form from the top right corner of the page.  Protection from insect pests was a major driving force of transgenic corn development.  Please note the environmental risk of decreasing diversity. Author Gewin is a freelance science journalist.
 
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5.7.4 Hawaiian Papaya   - Reading: Plant Management Network: Dennis Gonsalves and Steve Ferreira’s “Transgenic Papaya: A Case for Managing Risks of Papaya Ringspot Virus in Hawaii” Link: Plant Management Network:  Dennis Gonsalves and Steve Ferreira’s “Transgenic Papaya: A Case for Managing Risks of Papaya Ringspot Virus in Hawaii” (HTML)
 
Instructions: Please study this page.  Protection from viral infection was the major driving force of transgenic papaya development.  Please note the impact of the transgenic plant on the cultivation of nontransgenic papaya in the virus infected area.
 
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5.7.5 Rice   - Reading: GMO Compass: “Rice” Link: GMO Compass: “Rice” (HTML)
 
Instructions: Please study this page.  Please note that golden rice is an example of improved nutritional value of a genetically modified plant.
 
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5.8 Safety Issues   5.8.1 Human Health   - Reading: Ohio State University: Dresbach et al.’s “The Impact of Genetically Modified Organisms on Human Health” Link: Ohio State University: Dresbach et al.’s “The Impact of Genetically Modified Organisms on Human Health” (HTML or PDF)
 
Instructions: Please study this page.  You can access the PDF at the bottom of the page.
 
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5.8.2 Nontarget Organisms   - Reading: Emory University’s “GMO and Environment: Once a Gene’s In, Where Does It Go?” Link: Emory University’s “GMO and Environment: Once a Gene’s In, Where Does It Go?” (HTML)
 
Instructions: Please study this page.  Please note that a decreasing diversity makes species more vulnerable to environmental changes.
 
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