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Planta(2007) Institute of Botany, Chinese Academy of Science

Overexpression of OsCOIN, a putative cold inducible zinc finger protein, increased tolerance to chilling, salt and drought, and enhanced proline level in rice.

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Planta(2007) Institute of Botany, Chinese Academy of Science

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  1. Overexpression of OsCOIN, a putative cold inducible zinc finger protein, increased tolerance to chilling, salt and drought, and enhanced proline level in rice Overexpression of OsCOIN, a putative cold inducible zinc finger protein, increased tolerance to chilling, salt and drought, and enhanced proline level in rice Planta(2007) Institute of Botany, Chinese Academy of Science

  2. 1 2 3 4 Introduction Materials and methods Results Conclusion Contents Company Logo

  3. Introduction • Zinc finger proteins are one of transcriptional factors. • In plants zinc finger proteins are involved in growth, development and responses to environmental stresses. Company Logo

  4. Introduction • ABA as a phytohormone plays a critical role in response to various stress signals. • Numerous studies have shown that ABA is essential for the normal adaptive response to stress imposed by drought or high salinity. The endogenous ABA level increases under low-temperature conditions and application of exogenous ABA enhances cold tolerance of non-acclimatized plants. • Many of the biochemical and physiological changes under stress result from ABA-induced changes in gene expression patterns. Company Logo

  5. Introduction • The promoters of stress responsive genes have typical cis-regulatory elements and are regulated by various upstream transcriptional factors. These upstream transcription factors fall in the category of early genes and are induced within minutes of stress.The expression of the stress responsive genes leads to physiological responses. Company Logo

  6. Introduction • Proline has been reported to play roles in protecting enzymes from denaturation, stabilizing the machinery of protein synthesis, regulating the cytosolic acidity, increasing water-binding capacity, and acting as a reservoir of carbon and nitrogen source. Company Logo

  7. Isolation and characterization of OsCOIN gene • To isolate the cold-inducible gene in rice, the full-length OsCOIN cDNA was amplified by RT-PCR, cloned into pGEM-T easy and sequenced. Company Logo

  8. Subcellular localization • The OsCOIN cDNA was amplified with the primers. The PCR products were digested with XbaI and KpnI, and ligated with XbaI and KpnI double-digested pGFP221 to created pGFP-OsCOIN. The fusion construct and the GFP control vector were transformed into the onion epidermis cells by particle bombardment . Company Logo

  9. Subcellular localization Company Logo

  10. OsCOIN expression in various rice organs • The expression of the OsCOIN gene in various organs of the wild-type rice plants were analyzed by semi-quantitative RT-PCR using tubulin as an internal control. Company Logo

  11. OsCOIN expression in various rice organs Company Logo

  12. OsCOIN expression induction by cold Company Logo

  13. OsCOIN expression induction by ABA The expression of OsCOIN was induced by exogenous ABA suggesting that the expression of the OsCOIN is ABA-dependent in rice Company Logo

  14. OsCOIN expression induction by salt Company Logo

  15. OsCOIN expression induction by drought Company Logo

  16. Generation of OsCOIN transgenic rice • To explore its function, the OsCOIN gene driven by an ubiquitin promoter together with a GUS gene as a marker was transformed into rice plants by the Agrobacterium transformation approach. Company Logo

  17. Southernblot analysis Company Logo

  18. The levels of OsCOIN expression in wild type and OsCOIN transgenic rice Company Logo

  19. the effect of overexpression of OsCOIN gene on cold tolerance • To examine the effect of overexpression of OsCOIN gene on cold tolerance, rice seedlings of both wild type and OsCOIN transgenic rice were exposed to chilling temperature for 60, 72, and 84 h, and then returned to the normal growth conditions to allow their recovery. Company Logo

  20. the effect of overexpression of OsCOIN gene on cold tolerance Company Logo

  21. the effect of overexpression of OsCOIN gene on drought tolerance • To analyze the effect of overexpression of OsCOIN gene on tolerance to drought, rice seedlings of wildtype and the OsCOIN transgenic rice plants were submerged in the solutions containing 20% of PEG6000 for 12h and 24 h, and then returned to the normal growth conditions to allow their recovery. Company Logo

  22. the effect of overexpression of OsCOIN gene on drought tolerance Company Logo

  23. the effect of overexpression of OsCOIN gene on salt tolerance • To examine the effect of overexpression of OsCOIN gene on salt tolerance, the seeds of both wild type and OsCOIN transgenic rice were submerged in medium with 250 mM NaCl and germinated for 20 days. Company Logo

  24. the effect of overexpression of OsCOIN gene on salt tolerance Company Logo

  25. Expression of cold-responsive genes in OsCOIN transgenic rice OsP5CS was involved in the biosynthesis of proline Company Logo

  26. Determination of cellular proline levels Company Logo

  27. Conclusion • In this report we cloned and characterized OsCOIN, a novel cold-inducible gene in rice. OsCOIN, a putative zinc finger protein, is localized in both nuclear and cytoplasm membrane. Company Logo

  28. Conclusion • OsCOIN is expressed in all rice organs and strongly induced by low temperature, ABA, salt and drought. Overexpression of OsCOIN in transgenic rice significantly enhances tolerance to cold, salt and drought treatment. Company Logo

  29. Conclusion • Finally we showed that the expression of some of the known cold-responsive genes and the proline content of the cells were increased in the OsCOIN transgenic rice after the cold treatment. Company Logo

  30. Thank You ! www.themegallery.com

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