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General steps, Crop's genetic modification technique, desired DNA…

- - - - Infection of sterile in vitro explant
        
        culture
        
        screening via antibiotic(eg.kanomycin) or Reporter gene(eg. lac Z gene, GUS marker, GFP marker)
        
        adapt and culture
- - - - Agrobacterium rhizogenes-mediated transformation system
        
        Agent: Agrobacterium rhizogenes(Rhizobium rhizogenes)
        
        Soil born gram negative bacteria
        
        Transformation vector: Ri plasmid
        
        Infect on root wound region as phenolic acid will causes the bacteria approach the plant cell
        
        Causes hairy roots on many dicot plant and production of opines
        
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      - Agrobacterium tumeficens mediated transformation system
        
        Agent: Argobacterium tumeficiens
        
        Transformation vector: Ti plasmid
        
        infect in wound region
        
        Causes formation of tumor/ crown gall
  - - - Bombardment and shot plasmid DNA coated heavy metal into plant cell by gene gun.
        
        Advantage: fast, only way to modify the gene of monocot, suitable in non-dividing cells.
        
        Disadvantage: impact of heavy metal on plant cells
    - - Creation of transient pores in plasma lemma which allows the enter of DNA into the protoplast under a high field electric pulse
        
        Advantage: Low cell toxicity, convinient, fast, simple, efficient, allow the intake of large DNA(linear DNA rather than circular DNA)
        
        Disadvantage: Requirement of protoplast preparation
  - - - liposome(microscopic vesical caused by hydrated phospholipid) carries DNA and fused with targeted cell
        
        Disadvantage: Low efficiency and Laboriness, as not all plant suit
    - - Mixture of target plant cell and DNA-coated fibres (silicon carbide) causes the entrance of DNA into the cell due to collision
        
        Advantage: easy, fast, inexpensive procedures(x protoplast isolation)
        
        Disadvantage Low transformation efficiency(low survival after screening), silicon carbide damage to cell
  - - - Improve shelf life, phytoremediation, enhanced nutrient
    - - Indirect harm to other organism, allergenicity, gene transfer to other non targeted organism, unknown effect on human health.
        
        To reduce the adverse effect of GM crop on environment Environmental risk assessment is done
        
        Step1: Problem formulation, including hazard identification
        
        Step 2: Hazard characterization
        
        Step 3: Exposure characterization
        
        Step 4: Risk characterization
        
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    - - Less disruptive effect as compared to traditional breeding
      - GM foods failed to produce any negative health effect
    - - Herbicide resistance, BT gene, disease resistance/ cold draught resistance, fungal and pathogen tolerant
  - - - a) BT gene
      - b) inhibitor of protease