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상처에 의해서 유도되는 벼 calmodulin promoter의 transgenic 담배에서조직 특이적 발현

Tissue Specific Expression of Wound-Inducible RCaM-2 Promoter in Transgenic Tobacco Plants

  • 발행 : 2005.04.01

초록

Calmodulin 유전자의 발현 조절을 연구하기 위해, 벼 calmodulin promoter (RCaM-2)를 분리하여 GUS (report 유전자)에 융합하였다. GUS 활성은 정단조직, 근단 및 관다발 영역과 같은 성장조직에서 높게 발현되었다. 줄기와 페티올의 transverse 절단부위 GUS 활성은 관다발계의 안쪽에 제한되었으며 관다발계의 외부에 위치한 피층과 표피에서는 강하게 발현된 식물에서도 GUS 활성이 나타나지 않았다. GUS 활성은 어린 조직에서 특이적으로 발현되었으며 상처에 의해서 신속하게 증가하였다. RCaM-2 promoter는 세포분열이 왕성한 어린조직이나 생장점에서 강하게 발현되며 mechanical 신호에 의해서 현저히 유도되었다. 이러한 결과는 RCaM-2 유전자의 5'-flanking 영역이 상처에 의해서 유도되는 발현을 조절하는 것으로 추정된다.

To study calmodulin (CaM) gene expression and its regulation, rice CaM promoter (RCaM-2) was isolated and fused to $\beta-glucuronidase$ (GUS), reporter gene. X-Glue staining patterns revealed that GUS localization is high in meristemic tissues such as the stem apex, stolen tip, and vascular regions. GUS staining in the transverse sections of stem and petiole was restricted to the inside of the vascular system, and cortex and epidermis located outside of the vascular system usually did not show GUS staining even a plant that expressed strong activity. GUS activity was found to be tissue specific expressed and exhibited a dramatic transient increase in response to wounding. These results suggest that the 5'-flanking region of RCaM gene regulates wound-inducible expression.

키워드

참고문헌

  1. Bergey D. R. and C. A. Ryan. 1999. Wound- and systemininducible calmodulin gene expression in tomato leaves. Plant Mol. Biol. 40, 815-823 https://doi.org/10.1023/A:1006247624823
  2. Botella, J. R. and R. N. Arteca. 1994. Differential expression of two calmodulins in response to physical and chemical stimuli. Plant Mol. Biol. 24, 757-767 https://doi.org/10.1007/BF00029857
  3. Braam, J. and R. W. Davis. 1990. Rain-, wind-, touch-induced expression of calmodulin-related genes in Arabidopsis. Cell 60, 357-364 https://doi.org/10.1016/0092-8674(90)90587-5
  4. Braam, J. 1992. Regulated expression of the calmodulinrelated TCH genes in cultured Arabidopsis cells: Induction by calmodulin and heat shock. Proc. Natl. Acad. Sci. USA 89, 3213-3216
  5. Braam, J., M. L. Sistrunk, D. H. Polisensky, W. Xu, M. M. Purugganan, D. M. Antosiewicz, P. Campbell and K. A. Johnson. 1997. Plant responses to environmental stress: regulation and functions of the Arabidopsis TCH genes. Planta 203, 535-541
  6. Choi, Y. J., C. O. Lim, P. G. Shin, S. W. Gal, J. D. Bahk, G. An and M. J. Cho. 1993. Structural organization and sequence analysis of the calmodulin from rice (Oryza sativa cv, IR36). Mol. Cells 3, 255-261
  7. Choi, Y. J., E. K. Cho, S. I. Lee, C. O. Lim, S. W. Gal, M. J. Cho and G. An. 1996. Developmentally regulated expression of the rice calmodulin promoter in transgenic tobacco plants. Mol. Cells 6, 541-546
  8. Dong, A., H. Xin, Y. Yu., C. Sun, K. Cao and W.-H. Shen. 2002. The subcellular of an unusual rice calmodulin isoform, OsCaM61, depends on its prenylation ststua. Plant Mol. Biol. 48, 203-210 https://doi.org/10.1023/A:1013380814919
  9. Gawiennowski, M. C., D. Szyrnanski. L Y. Perera and R. E. Zielinski. 1993. Calmodulin isoforms in Arabidopsis encoded by multiple divergent mRNAs. Plant Mol. Biol. 22, 215-225 https://doi.org/10.1007/BF00014930
  10. Geiser, J. R, D. van Tuinen, S. E. Brockerhoff, M. M. Neff, and T. N. Davis. 1991. Can calmodulin function without binding calcium? Cell 65, 949-959 https://doi.org/10.1016/0092-8674(91)90547-C
  11. Heo, W. D., S. H. Lee, M. C. Kim, J. C. Kim, W. S. Chung, H. J. Chun, K. J. Lee, C. Y. Park, H. C. Park, J. Y. Choi and M. J. Cho. 1999. Involvement of specific calmodulin isoforms in salicylic acid-dependent activation of plant disease resistance responses. Proc. Natl. Acad. Sci. USA 96. 766-771
  12. Hoekema, A., P. R. Hirsch, P. J. Hooykaas and R. A. Schilperoort. 1983. A binary plant vector strategy based on separation of the Vie- and T-region of the Agrobacterium tumefaciens Ti plasmid. Nature (London) 303, 179-180 https://doi.org/10.1038/303179a0
  13. Jefferson, R. A. 1987. Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5, 387-405 https://doi.org/10.1007/BF02667740
  14. Jena, P. K., A. S. N. Reddy and B. W. Poovaiah. 1989. Molecular cloning and sequencing of a eDNA for plant calmodulin: Signal-induced changes in the expression of calmodulin. Proc. Natl. Acad. Sci. USA 86, 3644-3648
  15. Knight, H. 2000. Calcium signaling during abiotic stress in plants. Internal Reviews of Cytology 195, 269-324
  16. Koltunow, A. M. J. Truettner, K H. Cox, M. Wallroth and R. B. Goldberg. 1990. Different temporal and spatial gene expression patterns occur during anther development. Plant Cell. 2, 1201-1224 https://doi.org/10.1105/tpc.2.12.1201
  17. Lavine, A. R., I. Pennell, M. E. Alvarez, R. Palmer and C Lamb. 1996. Calcium mediated apoptosis in a plant hypersensitive disease resistance response. Current Biology 6, 427-436 https://doi.org/10.1016/S0960-9822(02)00510-9
  18. Lee, S. H., J. C Kim, M. S. Lee, W. D. Heo, H. Y. Seo, H. W. Yoon, J. CHong, S. Y. Lee, J. D. Bahk, I. Hwang and M. J. Cho. 1995. Identification of a novel divergent calmodulin isoform from soybean which has differential ability to activate calmodulin-dependent enzymes. J. Biol. Chem. 270, 21806-21812 https://doi.org/10.1074/jbc.270.37.21806
  19. Lee, S. H., H. Y. Seo, J. C Kim, W. D. Heo, W. S. Chung, K. J. Lee, M. C Kim, Y. H. Cheong, J. Y. Choi, C.O. Lim and M. J. Cho. 1997. Differential activation of NAD kinase by plant calmodulin isoforms. J. Biol. Chem. 272, 9252-9259 https://doi.org/10.1074/jbc.272.14.9252
  20. Lee, S. H., M. C. Kim, W. D. Heo, J. C. Kim, W. S. Chung, C. Y. Park, H. C. Park, Y. H. Cheong, C. Y. Kim, S.-H. Lee, K. J. Lee, J. D. Bahk, S. Y. Lee and M. J. Cho. 1999. Competitive binding of calmodulin isoforms to calmodulinbinding proteins: implication for the function of calmodulin isoforms in plants. Biochim. Biophys. Acta 1433, 56-67 https://doi.org/10.1016/S0167-4838(99)00149-1
  21. Ling, V. and R. E. Zielinski. 1989. Molecular cloning of eDNA sequences encoding the calcium-binding protein, calmodulin, from barley. Plant Physiol. 90, 714-719 https://doi.org/10.1104/pp.90.2.714
  22. Ling, V., I. Y. Perera and R. E. Zielinski. 1991. Primary structures of Arabidopsis calmodulin isoforms deduced from the sequences of cDNA clones. Plant Physiol. 96, 1196-1202 https://doi.org/10.1104/pp.96.4.1196
  23. Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bio assay with tobacco tissue cultures. Physiol Plant 15, 473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  24. Pandy, S., S. B. Tiwari, K C. Upadhyaya and S. K. Sopory. 2000. Calcium signaling: linking environmental signals to cellular functions. Crit. Rev. Plant Sci. 19, 291-318 https://doi.org/10.1016/S0735-2689(00)80006-4
  25. Perera, I. Y. and R. E. Zielinsk. 1991. Structure and expression of the Arabidopsis CaM-3 calmodulin gene. Plant Mol. Biol. 19, 49-64
  26. Perera, I. Y. and R. E. Zielinski. 1992. Synthesis and accumulation of calmodulin in suspension cultures of carrot (Daucus carota L.). Plant Physiol. 100, 812-819 https://doi.org/10.1104/pp.100.2.812
  27. Rodriguez-Concepcion, M., S. Yalovski, M. Zik, H. Fromm and W. Gruissem. 1999. The prenylation status of a novel plant calmodulin directs plasma membrane or nuclear localization of the protein. EMBO J. 18, 1996-2007 https://doi.org/10.1093/emboj/18.7.1996
  28. Sambrook, J., E. Fritsch. and T. Maniatis. 1989. Molecular Cloning : A Laboratory Mannual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
  29. Snedden, W. A. and H. Fromm. 1998. Calmodulin, calmodulin-related proteins and plant responses to the environment. Trends Plant Sci. 3, 299-304 https://doi.org/10.1016/S1360-1385(98)01284-9
  30. Takezawa, D., Z. H Liu, G. An and B. W. Poovaiah. 1995. Calmodulin genes family in potato: developmental and touch-induced expression of the messenger-RNA encoding a novel isoforms. Plant Mol. Biol. 27, 693-703 https://doi.org/10.1007/BF00020223
  31. Van Der Luit A. H., C. Olivari, A. Haley, M. R. Knight, A. J. Trewavas. 1999. Distinct calcium signaling pathways regulate calmodulin gene expression in tobacco. Plant Physiol. 121, 705-714 https://doi.org/10.1104/pp.121.3.705
  32. Xiao, C., H. Xin, H., A. W. Dong, C. R. Sun and K. M. Cao. 1999. A novel calmodulin-like protein gene in rice which has an unusual prolonged C-terminal sequence carrying a putative prenylation site. DNA Res. 6, 179-181 https://doi.org/10.1093/dnares/6.3.179
  33. Yamakawa, H., I. Mitsuhara, N. Ito, S. Seo. H. Kamada and Y. Ohashi. 2001. Transcriptionally and post-transcriptionally regulated response of 13 calmodulin genes to tobacco mosaic virus-induced cell death and wounding in tobacco plant. Eur. J. Biochem. 268, 3916-3929 https://doi.org/10.1046/j.1432-1327.2001.02301.x
  34. Yang, T., G. Segal, S. Abbo, M. Feldman and H. Fromm. 1996. Characterization of the calmodulin gene family in wheat: structure, chromosomal location, and evolutionary aspects. Mol. Gen. Genet. 252, 684-694 https://doi.org/10.1007/BF02173974
  35. Yang, T., S. Lev-Yadun, M. Feldman and H. Fromm. 1998. Developmentally regulated organ-, tissue, and cell-specific expression of calmodulin genes in common wheat. Plant Mol. Biol. 37, 109-120 https://doi.org/10.1023/A:1005902905512
  36. Zielinski, R. E. 1998. Calmodulin and calmodulin-binding proteins in plants. Annu. Rev. Plant Physiol. and Plant Mol. Biol. 49, 697-725 https://doi.org/10.1146/annurev.arplant.49.1.697