Challenges and Opportunities in Plant Biotechnology - PowerPoint PPT Presentation

Loading...

PPT – Challenges and Opportunities in Plant Biotechnology PowerPoint presentation | free to download - id: 741f68-M2JmM



Loading


The Adobe Flash plugin is needed to view this content

Get the plugin now

View by Category
About This Presentation
Title:

Challenges and Opportunities in Plant Biotechnology

Description:

Challenges and Opportunities in Plant Biotechnology Dave A. Vadnais – PowerPoint PPT presentation

Number of Views:110
Avg rating:3.0/5.0
Slides: 54
Provided by: Dave3184
Learn more at: http://faculty.nipissingu.ca
Category:

less

Write a Comment
User Comments (0)
Transcript and Presenter's Notes

Title: Challenges and Opportunities in Plant Biotechnology


1
Challenges and Opportunities in Plant
Biotechnology
  • Dave A. Vadnais

2
Lecture Outline
  • Applications in Plant Biotechnology
  • How Plants are Transformed
  • Functional Genomics
  • Recombinant DNA Techniques
  • Plant Transformation
  • Tissue Culture
  • Screening and Isolation of Transformants
  • Field and Laboratory Testing

3
Lecture Outline (cont.)
  • Gene Transfer to Commercial Cultivars
  • Product Commercialization
  • Plant Transformation Issues
  • Regulatory and Safety Issues
  • Societal Issues

4
Applications in Plant Biotechnology
  • Agronomic trait improvement
  • improved winter hardiness and stand persistence
  • insect resistance
  • pathogen tolerance
  • herbicide tolerance
  • nitrogen fixation
  • drought tolerance

5
Applications in Plant Biotechnology
  • Nutritional Improvement
  • increasing genistein content (soybeans)
  • oil quality (soybeans canola)
  • changing amino acid ratios (corn)
  • forage digestibility (alfalfa)
  • Improved Breeding
  • QTL cloning for grain productivity and plant
    height

6
Applications in Plant Biotechnology
  • Green Factories
  • biodegradable plastics from plants
  • pharmaceuticals from plants
  • Avian influenza vaccines (alfalfa)
  • Large scale pharmaceutical protein production
  • improved bioenergy sources
  • Biobutanol closer to gasoline than ethanol

7
Genetically Modified Organisms(GMOs)
  • All creatures are genetically modified
  • Genetic modification is natural
  • Viruses
  • Bacteria
  • Radiation

8
Genetically Modified Organisms(GMOs)
  • Methods used by people to genetically modify
    organisms
  • Chemical mutagens (Methyl Bromide)
  • Radiation (gamma radiation)
  • Genetic engineering

9
DNA to Protein
  • A Short Introduction

10
Protein Structure
11
Protein Structure
12
DNA Code is Degenerate
13
From DNA to Protein
Poly A
14
From DNA to Protein
15
The Plant Cell
  • Transit peptides direct proteins to organelles
  • mitochondria
  • chloroplast
  • vacuole
  • cell wall

16
How Plants are Transformed
  • What are the requirements?

17
Functional Genomics
  • Three major areas of study
  • Metabolomics
  • Understanding of products and the metabolic
    pathways used to their creation
  • Proteomics
  • Understanding the role of proteins in the life
    and regulation of living organisms
  • Genomics
  • Understanding the genetic control of various
    metabolic and proteomic pathways

18
Functional Genomics
  • Gene identification and isolation
  • micro-array technologies
  • cloning
  • sequencing
  • knowledge of regulatory elements

19
DNA Micro-Array Technology
  • Complete organism genome on micro array
  • Used to determine gene activity
  • Helps understanding of genetic pathways

20
Recombinant DNA Technologies
  • Cut DNA from Donor
  • Paste DNA into plasmid
  • Insert plasmid into Host Bacterium
  • Transform Plants

21
The Binary Vector
  • Multiplied in E. coli and Agrobacterium
  • Modular structure
  • Parts can be cut and pasted with great precision
  • Composed of DNA from many sources

22
Bacterial Selection
  • Bacteria are plated onto selection media
  • antibiotics
  • E. coli
  • DNA isolated for Particle Bombardment
  • Agrobacterium
  • Used to infect plant material

23
Plant Transformation
  • Four methods used today
  • Electroporation
  • Micro-injection
  • Particle Bombardment
  • Agrobacterium

24
Agrobacterium Transformation
25
Biolistic Plant Transformation
  • Biolistic gun
  • Patented by Dupont
  • One of the two main transformation methods used
  • Used highly compressed air

26
Biolistic Plant Transformation
  • Uses DNA coated gold or tungsten beads
  • Compressed air blasts beads into the cells or
    tissue
  • Cells take up the foreign DNA

27
Tissue Culture
  • The Big Picture
  • Regeneration from single cells

28
Plant Tissue Culture
  • Somatic Cell Embryogenesis
  • Cell Suspension Culture
  • Protoplast Regeneration
  • Anther Culture
  • Organogenesis

29
Somatic Embryogenesis
  • Donor plant and explants

30
Somatic Embryogenesis
  • Sterilization
  • Co-cultivation
  • Bombardment

31
Somatic Embryogenesis
  • Somatic Embryos Origin

32
Somatic Embryogenesis
  • Induction

33
Somatic Embryogenesis
  • Embryo Development

34
Somatic Embryogenesis
  • Germination

35
Somatic Embryogenesis
  • Plant Development

36
Screening and Isolation of Transformants
  • Polymerase Chain Reaction (PCR)
  • is the gene present?
  • Southern Analysis
  • gene copy number
  • Northern Analysis
  • gene expression patterns
  • Enzyme Assays
  • gene product activity

37
Polymerase Chain Reaction(PCR)
  • Only Foreign DNA targeted
  • Only specific regions of DNA amplified
  • Easy and quick to screen 100s of plants

38
Southern Analysis
  • Determines gene copy number
  • Detects only foreign DNA

39
Laboratory Testing of Transgenic Plants
  • Laboratory and Greenhouse Testing
  • Is the plant producing the required material?
  • Is it healthy?
  • Will it survive?

40
Field Testing of Transgenic Plants
  • Field Testing
  • Allows evaluation of the new plant in the real
    world
  • Will the plant survive real world stresses?
  • Is the new trait inherited?
  • Will the trait transfer to commercial varieties?

41
Plant Biotech Products
  • Golden Rice
  • Gene insertion (ProVitamin A and Phytase)
  • Enhanced beta-carotene (vitamin A) and Iron
    absorption
  • Helps prevent Vitamin A deficiency (VAD) and Iron
    Deficiency Anemia (IDA)

42
Plant Biotech Products
  • Flavr savr tomato
  • Gene knockout (antisense expression)

43
Plant Biotech Products
  • Bt Corn
  • Gene overexpression (constitutive)

44
Plant Biotech Products
  • RR Soybean
  • Gene overexpression (constitutive)

45
Plant Biotech Products
  • Salt tolerant tomato
  • Gene insertion
  • Gives ability to grow on salt contaminated soils
  • Ability to accumulate salt from the soil
  • Zhang and Blumwald (2001) Nature Biotechnology 19

46
Plant Biotech Products
  • High Energy Alfalfa
  • Gene targeting
  • Yeast invertase to vacuole
  • using patatin transit peptide

47
Commercialization Issues
  • Freedom To Operate
  • Many of the enabling technologies patented
  • May require permission and/or payment of
    licensing fees
  • Can cause many problems in getting the product to
    the market
  • Could force researchers to re-invent the wheel

48
Major Concerns About Plant Biotechnology
  • Imprecise technology
  • Food and product safety
  • Cross-species transfer (horizontal gene transfer)
  • Genetic pollution
  • New weeds

49
Imprecise Technology
  • Scientists can now place a single copy of a gene
    precisely in the genome
  • Matrix Attachment Regions
  • More stable expression of transgenes
  • Cre-Lox and FLP/FRT system
  • Enables precise placement and removal of
    transgenes
  • Antibiotic resistance genes
  • Herbicide resistance genes
  • Transgenic trait genes

Ow and Srivastava, 2004 Trends in Biotech 22
(12) 627-629
50
Food and Product Safety
  • Canadian Food Inspection Agency Roles
  • Regulate transgenic field trials
  • Ensures field trials conducted in reproductive
    isolation
  • Environmental Protection
  • prevent gene pollution (gene transfer to
    non-target species)
  • prevent the environmental release of toxins
  • prevent the production of Super-weeds

51
Food and Product Safety
  • Canadian Food Inspection Agency Roles
  • Ensure Food Safety
  • product is non-toxic to humans or animals
  • must be equivalent to current food products
  • must be non-allergenic

52
Food and Product Safety
  • Codex Alimentarius Commission
  • Established in 1963 by WHO and FAO of the UN
  • 2003 - established risk/safety assessment
    guidelines for food derived from genetically
    modified plants and micro-organisms
  • Serves as a reference for international trade
    disputes over the safety of internationally
    traded foods

53
Questions?
About PowerShow.com