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Questions and Answers
Which method of plant genetic engineering involves treating protoplasts with Polyethylene Glycol to induce macro DNA uptake?
Reporter genes such as GFP and GUS are selectable markers in plant genetic engineering.
False
What is the purpose of the GFP (Green Fluorescent Protein) reporter gene in plant genetic engineering?
To make plant organs light up green when interacting with UV light.
The first FDA-approved GMO tomato on the market in 1984 had enhanced ripening due to the polygalacturonase gene, which was suppressed using __________.
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Match the following plant genetic engineering delivery methods with their descriptions:
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Which method allows for direct transfer of one or a few genes between closely or distantly related species?
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Reporter genes such as GFP and GUS are selectable markers.
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What does GUS stand for in the context of plant genetics?
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The first FDA-approved GMO tomato on the market enhanced tomato ripening by suppressing the expression of Polygalacturonase using ____.
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What is one of the techniques used in plant genetic engineering that involves treating protoplasts with Polyethylene Glycol to induce macro and DNA uptake?
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Reporter genes like GFP and GUS are selectable markers for plant genetic engineering.
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What was the first FDA-approved GMO tomato on the market and what enhancement did it provide?
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Golden Rice was transformed with 3-beta carotene synthesis genes using an ___________-specific promoter.
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Match the plant genetic engineering tool with its description:
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Study Notes
Plant Genetic Engineering
- Humans have been manipulating plant genes for 10,000 years through selective breeding
- Agriculture originated in the Mediterranean
- Maize was developed from teosinte through selective breeding, taking 700 years
Conventional Breeding
- Limited to exchanges between same/related species
- No guarantee of obtaining specific gene combinations
- Undesirable genes can be transferred with desirable ones
- Long time to achieve desired results
Genetic Engineering
- Allows for direct transfer of one or a few genes between closely or distant species
- Can be achieved in a shorter amount of time
Tools/Techniques of Plant Genetic Engineering
- Polyethylene Glycol (PEG): treats protoplasts to induce DNA uptake by changing cell permeability
- Liposomes: foreign DNA is encapsulated and protoplasts engulf liposomes in presence of PEG via endocytosis
- Electroporation: foreign DNA is taken up by protoplasts in a high voltage electric field
- Whisker-mediated: silicon carbide fibers and protoplasts are vortexed, creating holes in the plasma membrane for DNA uptake
- Microinjection: foreign DNA is injected into plant protoplasts with glass needles
- Laser Microbeam: UV laser creates holes in the protoplast to introduce foreign DNA
- Desiccation: dehydrated embryos can absorb foreign DNA when rehydrated with DNA solution
Reporter Genes
- GFP (Green Fluorescent Protein): cloned from jellyfish, interacts with UV light to produce a green glow
- GUS (E. coli's B-glucuronidase): interacts with X-Gluc and K+ferricyanide/ferrocyanide mix to turn blue
History of GMOs
- 1984: First FDA-approved GMO tomato on the market, used antisense to suppress polygalacturonase expression for enhanced tomato ripening
- 1992-2000: Golden Rice, used endosperm-specific promoter to transform 3-beta carotene synthesis genes for vitamin A deficiency
Plant Biotechnology Applications
- Medicine
- Disease/pest resistance
- Drought tolerance
- Biofuel
- Nutrition
- High yield
- Comparative genomics
- Phytosensor (detecting toxins) and phytoremediation (removing chemicals from soil)
Additional Notes
- Tobacco plants were genetically modified to produce "Plantibodies" that inactivated Ebola
- Every gene must have a promoter, unless it's an operon
- Promoter -> GUS -> Terminator -> mRNA -> Protein
- Reporters (GUS, GFP, LUC) are not selectable markers
- Gene Gun/agrobacterium transformation are most popular methods to transfer foreign genes to plants
- Delivery system -> integration -> recovery, with PCR necessary to confirm integration
Plant Genetic Engineering
- Humans have been manipulating plant genes for 10,000 years through selective breeding
- Agriculture originated in the Mediterranean
- Maize was developed from teosinte through selective breeding, taking 700 years
Conventional Breeding
- Limited to exchanges between same/related species
- No guarantee of obtaining specific gene combinations
- Undesirable genes can be transferred with desirable ones
- Long time to achieve desired results
Genetic Engineering
- Allows for direct transfer of one or a few genes between closely or distant species
- Can be achieved in a shorter amount of time
Tools/Techniques of Plant Genetic Engineering
- Polyethylene Glycol (PEG): treats protoplasts to induce DNA uptake by changing cell permeability
- Liposomes: foreign DNA is encapsulated and protoplasts engulf liposomes in presence of PEG via endocytosis
- Electroporation: foreign DNA is taken up by protoplasts in a high voltage electric field
- Whisker-mediated: silicon carbide fibers and protoplasts are vortexed, creating holes in the plasma membrane for DNA uptake
- Microinjection: foreign DNA is injected into plant protoplasts with glass needles
- Laser Microbeam: UV laser creates holes in the protoplast to introduce foreign DNA
- Desiccation: dehydrated embryos can absorb foreign DNA when rehydrated with DNA solution
Reporter Genes
- GFP (Green Fluorescent Protein): cloned from jellyfish, interacts with UV light to produce a green glow
- GUS (E. coli's B-glucuronidase): interacts with X-Gluc and K+ferricyanide/ferrocyanide mix to turn blue
History of GMOs
- 1984: First FDA-approved GMO tomato on the market, used antisense to suppress polygalacturonase expression for enhanced tomato ripening
- 1992-2000: Golden Rice, used endosperm-specific promoter to transform 3-beta carotene synthesis genes for vitamin A deficiency
Plant Biotechnology Applications
- Medicine
- Disease/pest resistance
- Drought tolerance
- Biofuel
- Nutrition
- High yield
- Comparative genomics
- Phytosensor (detecting toxins) and phytoremediation (removing chemicals from soil)
Additional Notes
- Tobacco plants were genetically modified to produce "Plantibodies" that inactivated Ebola
- Every gene must have a promoter, unless it's an operon
- Promoter -> GUS -> Terminator -> mRNA -> Protein
- Reporters (GUS, GFP, LUC) are not selectable markers
- Gene Gun/agrobacterium transformation are most popular methods to transfer foreign genes to plants
- Delivery system -> integration -> recovery, with PCR necessary to confirm integration
Plant Genetic Engineering
- Plant genes have been manipulated for 10,000 years through selective breeding.
- Conventional breeding is limited to exchanges between same/related species and can take a long time to achieve desired results.
- Genetic engineering allows for direct transfer of one or a few genes between closely or distant species in a shorter amount of time.
Tools and Techniques of Plant Genetic Engineering
- Polyethylene Glycol (PEG) is used to treat protoplasts, inducing macro and DNA uptake by changing cell permeability.
- Liposomes are used to encapsulate foreign DNA, which is then engulfed by protoplasts in the presence of PEG via endocytosis.
- Electroporation is used to take up foreign DNA by protoplasts in a high voltage electric field.
- Whisker-mediated method uses silicon carbide fibers to create holes in the plasma membrane, allowing DNA to enter the cell.
- Microinjection involves injecting foreign DNA into plant protoplasts with glass needles.
- Laser Microbeam uses a UV laser to create holes in protoplasts to introduce foreign DNA.
- Desiccation involves dehydrated embryos absorbing foreign DNA when rehydrated with a DNA solution.
Reporter Genes
- GFP (Green Fluorescent Protein) is cloned from jellyfish and interacts with UV light to produce a green glow.
- GUS (E. coli's B-glucuronidase) interacts with X-Gluc and a K+ ferricyanide/ferrocyanide mix to produce a blue color.
GMO Applications
- The first FDA-approved GMO tomato was released in 1984, which used antisense to suppress the expression of polygalacturonase to enhance tomato ripening.
- Golden Rice was developed from 1992 to 2000, using an endosperm-specific promoter to transform 3-beta carotene synthesis genes, which could help populations with vitamin A deficiency.
Plant Biotechnology Applications
- Medicine: Genetic modification can be used to produce "plantibodies" like those used to inactivate Ebola.
- Disease/Pest Resistance: Genetic modification can be used to enhance disease resistance in plants.
- Drought Tolerance: Genetic modification can be used to enhance drought tolerance in plants.
- Biofuel: Genetic modification can be used to produce biofuels.
- Nutrition: Genetic modification can be used to enhance nutritional content in plants.
- High Yield: Genetic modification can be used to enhance crop yields.
- Comparative Genomics: Genetic modification can be used to study comparative genomics.
- Phytosensor/Phytoremediation: Genetic modification can be used to detect toxins and remove chemicals from soil.
Gene Transfer Methods
- Gene Gun: A high velocity particle delivery system that uses helium to rupture a disk, propelling gold-covered DNA into target cells.
- Agrobacterium Transformation: Uses a disarmed plasmid to transfer DNA into plant cells, which is then integrated into the host genome.
- PCR is necessary to confirm integration of foreign genes into the host genome.
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Description
Discover the history of plant genetic manipulation and the differences between genetic engineering and conventional breeding techniques.
