Tanaman Transgenik

1. TanamanTransgenik Fifi Fitriyah Masduki 12 November 2010

2. Transgenics are a Biotechnology Product Biotechnology - General Definition The application of technology to improve a biological organism Biotechnology - Detailed Definition The application of the technology to improve the biological function of an organism by adding genes from another organisms Sumber : Phil McClean, NDSU, 2003

3. What About the Term Genetic Engineering? Genetic engineering is the basic tool set of biotechnology Genetic engineering involves: • Isolating genes • Modifying genes so they function better • Preparing genes to be inserted into a new species • Developing transgenes Sumber : Phil McClean, NDSU, 2003

4. What is a transgenic? Transgene – the genetically engineered gene added to a species Transgenic – an organism containing a transgene introduced by technological (not breeding) methods GMO- an organism that expresses traits that result from the introduction of foreign DNA Originally a term equivalent to transgenic organism

5. Why create transgenic plant? Increase yield Enhance taste Enhance Nutrition

6. Molecular Farming Why create transgenic plant? TembakauvsE. coli • 1 tobacco plant = million seeds • The leaves of 1 tobacco plant produce more recombinant protein than a 300-liter fermenter in E. coli • It cost about 50x more to produce 1kg of a recombinat protein in E. coli than in trangenic plants

7. The Golden Rice Story • Vitamin A deficiency is a major health problem • Causes blindness • Influences severity of diarrhea, measles • >100 million children suffer from the problem • For many countries, the infrastructure doesn’t exist to deliver vitamin pills • Improved vitamin A content in widely consumed cropsanattractive alternative

8. IPP Geranylgeranyl diphosphate Phytoene synthase Phytoene Problem: Rice lacks these enzymes Phytoene desaturase ξ-carotene desaturase Lycopene Lycopene-beta-cyclase Normal Vitamin A “Deficient” Rice  -carotene (vitamin A precursor) -Carotene Pathway Problem in Plants

9. IPP Geranylgeranyl diphosphate Phytoene synthase Phytoene Vitamin A Pathway is complete and functional Phytoene desaturase ξ-carotene desaturase Lycopene Lycopene-beta-cyclase  -carotene (vitamin A precursor) Golden Rice The Golden Rice Solution -Carotene Pathway Genes Added Daffodil gene Single bacterial gene; performs both functions Daffodil gene

10. The Roundup Ready Story • Glyphosate is a broad-spectrum herbicide • Active ingredient in Roundup herbicide • Kills all plants it come in contact with • Inhibits a key enzyme (EPSP synthase) in an amino acid pathway • Plants die because they lack the key amino acids • A resistant EPSP synthase gene allows crops • to survive spraying

11. Shikimic acid + Phosphoenol pyruvate Plant EPSP synthase 3-Enolpyruvyl shikimic acid-5-phosphate (EPSP) Aromatic amino acids Roundup Sensitive Plants + Glyphosate X X Without amino acids, plant dies X X

12. Roundup Resistant Plants Shikimic acid + Phosphoenol pyruvate + Glyphosate RoundUp has no effect; enzyme is resistant to herbicide Bacterial EPSP synthase 3-enolpyruvyl shikimic acid-5-phosphate (EPSP) With amino acids, plant lives Aromatic amino acids

13. Insect resistant plant story • Insect resistant cotton – Bt toxin kills the • cotton boll worm • transgene = Bt protein Source: USDA • Insect resistant corn – Bt toxin kills the • European corn borer • transgene = Bt protein Normal Transgenic

17. Introducing the Gene or Developing Transgenics Steps 1. Create transformation cassette 2. Introduce and select for transformants

18. 1. Gene of interest • The coding region and its controlling elements 2. Selectable marker • Distinguishes transformed/untransformed plants 3. Insertion sequences • Aids Agrobacterium insertion Transformation Cassettes Contains

19. Tissue must be capable of developing into normal plants • Leaf, germinating seed, immature embryos • Develop shoots • Root the shoots Transformation Steps Prepare tissue for transformation Introduce DNA • Agrobacterium or gene gun Culture plant tissue Field test the plants • Multiple sites, multiple years

20. Agrobacterium Tissue culture required to generate transgenic plants • Gene Gun Delivering the Gene to the Plant • Transformation cassettes are developed in the lab • They are then introduced into a plant • Two major delivery methods

21. Genome organization in plant

22. Chloroplast vs Nuclear

23. Chloroplast vs Nuclear

24. Chloroplast vs Nuclear

25. Chloroplast vs Nuclear

26. Chloroplast vs Nuclear WT Flowers + transgenic pollen Transgenic Flowers + WT pollen

27. Resistance shoot Heteroplasmy Homoplasmy Chloroplast transformation (Source: Bock and Khan 2004)

30. Edible Vaccines Transgenic Plants Serving Human Health Needs • Works like any vaccine • A transgenic plant with a pathogen protein gene is developed • Potato, banana, and tomato are targets • Humans eat the plant • The body produces antibodies against pathogen protein • Humans are “immunized” against the pathogen • Examples: • Diarrhea • Hepatitis B • Measles

31. .. Thanks for your attention!!

32. Basic Science Purpose

34. Two Hormones Affect Plant Differentiation: Auxin: Stimulates Root Development Cytokinin: Stimulates Shoot Development Generally, the ratio of these two hormones can determine plant development: Auxin↓Cytokinin = Root Development Cytokinin↓Auxin = Shoot Development Auxin = Cytokinin = Callus Development Basis for Plant Tissue Culture