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Rice biotechnology


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Rice biotechnology

  1. 1. Rice(Oryza sativa) is one of the most important cereal crops, providing staple food for nearly one-half of the global population. More than 90% of rice is grown and consumed in Asia where about 55% of the world’s population lives, reflecting the value of rice in daily human life. Its importance can be estimated by the fact that the year 2004 was declared as International Year of Rice by the United Nations Food and Agriculture Organization. In direct proportion to the predicted rise in the world’s human population, rice consumption and demand will increase over the next several decades.
  2. 2. 1.To improve yield 2.Little land is now available for farming 3.Crops subjected to environmental stress 4.To enhance the nutritional value of food we eat
  3. 3. 1. Agrobacterium- mediated Gene Transfer • CAT1 intron was inserted into the selectable marker gene for hygromycin resistance • Several useful traits that uses Ag transformation are: abiotic stress tolerance, biotic stress tolerance, herbicide tolerance, nutritional enhancement and enhanced photosynthesis
  4. 4.  Isolate gene that direct cells to make protein of interest  Attach the gene to the promoter that works in plant  Insert the promoter-gene and a gene for selectable marker into plant cells
  5. 5. 2. Particle Bombardment/ Biolistics • considered genotype independent and less labor intensive. • associated sometimes with some risk due to arrangement of multiple copies of transgenes, particularly in the form of inverted repeats and problem of high copy number of the transgene, unlike Agrobacterium- mediated transformation. • has been used for investigation on promoter, stress tolerance, nutritional enhancement, gene expression, plant development and grain yield
  6. 6. 3. PEG (polyethylene glycol) Mediated transformation • Rice protoplasts can be transformed with naked DNA by treatment with PEG in the presence of divalent cations such as calcium. • Toriyama et al. (1988) and Zhang and Wu (1988) were the first to recover transgenic rice using PEG technology and thus it is the first technique used in transgenic rice production
  7. 7. 4. Electroporation • Shimamoto et al. (1989) were the first to recover fertile transgenic plants using electroporation in japonica rice. • protoplasts are not easy to work with and regeneration of fertile plants is problematic. • a single-cell manipulation supporting robot for high throughput microinjection of rice protoplasts have been developed
  8. 8. 5. In planta transformation • To avoid tissue culture and sterile conditions, in planta transformation method that depends on a needle dipped in Agrobacterium culture to prick the seed‘s embryonic portion that subsequently grows into a plant and sets transgenic seeds were developed. Supartana et al. (2005)
  9. 9. A.Stress Tolerance 1. Biotic Stress Tolerance a. Insect resistance- • By introducing the gene for virus-enhancing factor in rice, the effectiveness of baculoviral insecticides against feeding armyworm larvae was enhanced. (Hakuhara et al,,1999)
  10. 10. Striped stemborer Rice leaffolder
  11. 11. • Another strategy for insect resistance is the use of plant proteinase inhibitor. Earlier, Irie et al. (1996) generated transgenic rice resistant to insect storage pests using hydrolase inhibitors.
  12. 12. b. Bacterial Disease Resistance- • Bacterial blight, caused by Xanthomonas oryzae pv. Oryzae (Xoo), along with blast and sheath blight are most important • The most promising endogenous rice gene for bacterial blight resistance identified so far is Xa21 (Song et al., 1995), which conferred complete protection against bacterial blight. Cecropins are antibacterial peptides having broad spectrum bacteriallytic activity against gram negative and gram positive bacteria, but not against eukaryotic cells.
  13. 13. c. Fungal Disease Resistance • Most promising endogenous resistance (R) gene, Pi-ta (cytoplasmic NBS receptor), is responsible for resistance to fungal diseases. • Another gene, OSDR8, is involved in thiamine biosynthesis and acts upstream in defense signal transduction pathway. Pathogenesis-related (PR) genes have been used widely to address fungal tolerance in plants.
  14. 14. d. Viral Resistance- • Insect-vectored viral diseases cause considerable damage to the rice plant and drastically reduce the yield of the plant. • To generate virus-resistant transgenic rice, japonica rice was transformed with a hammerhead ribozyme. • Resistance against this virus has been introduced in rice by expressing 39 kDa spike protein of RRSV. RHBV is the causative virus of a major rice viral disease that can cause significant damage (up to 50% loss) of the total yield.
  15. 15. 2. Abiotic Stress Resistance- • responsible for most of the reduction that differentiates yield potential from harvestable yield. • high temperature, chilling, freezing, water deficit (drought and salinity), high light intensity, flooding, and exposure to ozone and heavy metals. • Overproduction of various compatible solutes has been tested in rice, e.g., glycine betaine, trehalose, proline and polyamines, to achieve significant drought, cold, and salt tolerance.
  16. 16. • Water channel proteins, aquaporins, are members of the major intrinsic protein family which regulate the passive movement of water across membranes. Aquaporins might have a possible role in providing drought tolerance to plants as they affect the root water uptake. • A promising strategy against salinity stress is the use of Na+ transporters, which transport cytosolic Na+ to vacuole and, thus protect cellular machinery. Na+/H+ antiporters from Artiplex or E.coli confer high salt tolerance in transgenic rice (Ohta et al., 2002).
  17. 17. 3. Herbicide Tolerance- • In rice, bar was the first gene tested for herbicide (Basta, glufosinate) tolerance. Two gene families that play major roles in conferring tolerance to herbicides are P450 monooxygenase and glutathione S-transferase. • Cytochrome P450 monooxygenase, a drug-metabolizing enzyme system in plants and animals, has been widely used to address herbicide tolerance trait in rice. • transgenic plants showed tolerance to sulfonylurea herbicide chlorsulfuron
  18. 18. 1. Nutritional Enhancement >> ‗Golden Rice‘ with provitamin A was generated. Later, the entire pathway for β-carotene biosynthesis was reconstituted by using just two structural genes, PSY and crt1
  19. 19. IRON FORTIFIED >> Human lactoferrin (HLF) is a major iron binding glycoprotein in breast milk. Transgenic rice accumulating HLF in seed provided a novel means for nutrient supplement for infants as recombinant human lactoferrin could maintain the biological activity in transgenic rice seeds. (Nandi et al., 2002)
  20. 20. 2. Alteration of Starch Content • In vivo modification of starch using genetic engineering • Wheat puroindoline genes PINA and PINB were introduced in rice to modify the grain texture. Transgenic grains were very soft in texture and referred as ―Soft Rice.‖ (Nakamura et al., 2002)
  21. 21. Rice is a diverse crop that grows in different ecosystems. Current gene evolution should provide wide scope for the application of biotechnology across ecosystems and crop barriers. It is desirable to create superior transgenic rice plants that can grow in compromised environments and have higher yields with decreasing arable land availability. Gene pyramiding or multigene engineering using genes involved in various agronomic traits is a powerful approach to obtain superior rice varieties (Ashikari and Matsuoka, 2006). Indeed, rice biotechnology gives great benefit and safety to all people—particularly resource-poor people.
  22. 22. Bhullar, N. K., & Gruissem, W. (2013). Nutritional enhancement of rice for human health : The contribution of biotechnology. Biotechnology Advances, 31(1), 50–57. doi:10.1016/j.biotechadv.2012.02.001 Brown, D. C. W., & Thorpe, T. A. (1995). Crop improvement through tissue culture, II, 409– 415. Chen, H. (n.d.). Development of Transgenic Insect- resistance ( IR ) Rice Rice insect pests. DNA Technology & the Story of ―Golden Rice‖. (n.d.). Farooq, S. (n.d.). Women in rice biotechnology: success that will have an impact in the times ahead. Gangwar, B. (2010). Strategies for improving production in rice based cropping systems. Kathuria, H., Giri, J., Tyagi, H., & Tyagi, A. K. (2007). Advances in Transgenic Rice Biotechnology, 65–103. doi:10.1080/07352680701252809 Redona, E. D. (2004). Rice Biotechnology for Developing Countries in Asia, 201–232. Xie, F. (n.d.). Hybrid Rice Breeding & Seed Production. Retrieved from Zhu, Z., & Wu, R. (2008). Regeneration of transgenic rice plants using high salt for selection without the need for antibiotics or herbicides, 174, 519–523. doi:10.1016/j.plantsci.2008.01.017