Application of PCR

1
Generation of Transgenic Plants
leaf
single cell
transformation of cell (gene transfer)
protoplast transformation
gene gun
Agrobacterium tumefaciens
regeneration
transformed cell
callus culture
transgenic plant
2
Cultivation of Transgenic Plants in 2005
3
Gene Transfer by Gene Gun (Biolistic)
4
Gene Transfer by Gene Gun in Plant Cells
pressure compartment
suppport for DNA-coated particles
breaking plate
DNA-coated particles
scattering grid
plant tissue in petri dish
5
Generation of Transgenic Plants
leaf
single cell
transformation of cell (gene transfer)
protoplast transformation
gene gun
Agrobacterium tumefaciens
regeneration
transformed cell
callus culture
transgenic plant
6
Generation of Transgenic Plants
leaf
single cell
transformation of cell (gene transfer)
protoplast transformation
gene gun
Agrobacterium tumefaciens
regeneration
transformed cell
callus culture
transgenic plant
7
RoundupReady Soy Beans (RRS)

• Soy beans are processed to soy oil and bruised
  grain
• 30.000 food products contain soy oil (chips, ice
  cream, bakery products, candy bars...)
• RoundupReady soy beans are resistant against the
  herbicide Roundup, glyphosate,
  N-(phosphonomethyl)-glycine) sold by Monsanto
• Glyphosate inhibits the enzyme EPSPS
  (3-enolpyruvyl-shikimate-5-phosphate-synthase)
  which is needed for the biosynthesis of aromatic
  amino acids
• Cells die without aromatic amino acids

8
Glyphosate - Mode of Action -
9
RoundupReady Soy Beans (RRS)

• Soy beans are processed to soy oil and bruised
  grain
• 30.000 food products contain soy oil (chips, ice
  cream, bakery products, candy bars...)
• RoundupReady soy beans are resistant against the
  herbicide Roundup, glyphosate,
  N-(phosphonomethyl)-glycine) sold by Monsanto
• Glyphosate inhibits the enzyme EPSPS
  (3-enolpyruvyl-shikimate-5-phosphate-synthase)
  which is needed for the biosynthesis of aromatic
  amino acids
• Cells die without aromatic amino acids
• RoundupReady soy beans contain a
  glyphosate-resistant bacterial gene from
  Agrobacterium tumefaciens CP4
• 30 reduction of use of glyphosate
• RoundupReady soy beans is sold by Monsanto
• USA produces 60 million tons of soy beans per
  year (50 of worldwide harvest)
• ---------------------

10
Vector for Glyphosate Resistance
11
T-DNA from Agrobacterium tumefaciens
gene of interest
plant promoter
plant resistance gene for selection
terminator
left border of T-DNA
right border of T-DNA
bacterial resistance gene for selection
ori /origin of replication
bacterial promoter
12
Labeling

• Food, food ingredients and food supplement have
  to be labeled when they are GMO, consist of GMO
  or contain material of GMO
• Labeling is required when the percentage of GMO
  material is greater than 0.9 in food containing
  a mixture of normal and GMO material
• Examples
• Food or food supplement are GMOs GM tomato, GM
  patato, GM corn, GM fish...
• Food or food supplement contain GMOs GM bacteria
  in yogurt, GM yeast in beer...
• Food or food ingredients are derived from GMO
  oil from GM soy beans or GM corn, sugar form GM
  sugar beet, starch from GM corn...
• Food or food supplement are isolated from GMO
  Vitamin C, vitamin B2, glutamate...
• Labeling is not required when
• meat, eggs or milk comes from animals fed with
  GMO plants
• Material from GMO fed to microorganisms, e.g.
  yeast cells fed with starch from GM corn
• enzymes isolated from GMO chymosin produced by
  GM bacteria
• ---------------------------------------------
• GMO genetically modified organism

13
End