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RICE BIOTECHNOLOGY Presented by: Janine V. Samelo BSChem4

Rice biotechnology

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Page 1: Rice biotechnology

RICE BIOTECHNOL

OGYPresented by:

Janine V. SameloBSChem4

Page 2: Rice biotechnology

Introduction: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 directproportion to the predicted rise in the world’s human population,rice consumption and demand will increase over the next severaldecades.

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Why the need for Biotechnology?

1.To improve yield2.Little land is now available for farming3.Crops subjected to environmental stress4.To enhance the nutritional value of food we

eat

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Methods of transformation1. 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

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Methods of transformation

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

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Methods of transformation2. 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

 

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Methods of transformation3. 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

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Methods of transformation

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  

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Methods of transformation

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)

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Biotech Applied to Rice

A.Stress Tolerance1. Biotic Stress Tolerancea. 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)

 

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Biotech Applied to Rice

Striped stemborer Rice leaffolder

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Biotech Applied to Rice

• 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.

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Biotech Applied to Riceb. 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.

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Biotech Applied to Ricec. 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.

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Biotech Applied to Rice

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.

 

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Biotech Applied to Rice2. 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.

 

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Biotech Applied to Rice• 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).

 

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Biotech Applied to Rice3. 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

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Improvement of grain quality1. 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 

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Improvement of grain quality

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)

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Biotech Applied to Rice

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)

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Gene modification

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Conclusion: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.

 

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THANK YOU for listening !!

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References: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:10.1080/07352680701252809Redona, E. D. (2004). Rice Biotechnology for Developing Countries in Asia, 201–232.Xie, F. (n.d.). Hybrid Rice Breeding & Seed Production. Retrieved from [email protected] 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