Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
권호 표지
DOI QR코드

DOI QR Code

Introduction and Expression of a Thaumatin-like Protein from Rice in American Ginseng Following Agrobacterium-mediated Transformation

  • Chen, W.P. (Department of Biological Sciences, Simon Fraser University) ;
  • Punja, Z.K. (Department of Biological Sciences, Simon Fraser University)
  • Published : 2003.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Agrobacterium-mediated transformation of American ginseng (Panax quinquefolius L.) with strain LBA 4404 containing a rice thaumatin-like protein gene is described. The selectable markers used were phosphinothricin acetyltransferase and hygromycin phosphotransferase genes. Epicotyl explants from seedlings were precultured for 5-7 days on Murashige and Skoog medium with ${\alpha}$-naphthaleneacetic acid and 2,4 dichlorophenoxyacetic acid at 10 ${\mu}$M and 9 ${\mu}$M, respectively (ND medium), prior to Agrobacterium infection. The explants were immersed in a bacterial suspension for 20 min. A post-infection co-culture period of 3-4 days was provided on ND medium. Selection for transformed calli was conducted on ND medium with 20 mg/L phosphinothricin followed by 100 mg/L hygromycin over an 8-month period. it transformation frequency of 24.8% was achieved at the callusing phase. The presence of the transgenes in calli was confirmed by Southern hybridization and polymerase chain reaction analysis. The expression of the thaumatin-like protein gene in ginseng calli was demonstrated by Western blot analysis. Somatic embryos were produced from both transgenic calli and suspension cultures, and plantlets were recovered that expressed the transgenic thaumatin-like protein gene.

Keywords

References

  1. Bradford, M. M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72, 248-254 (1976) https://doi.org/10.1016/0003-2697(76)90527-3
  2. Casas, A. M., Cereal transformation through particle bombardment in Janick, J. (ed), Plant breeding reviews, vol. 13.: John Wiley & Sons Inc., New York, pp 235-264 (1995)
  3. Chen, W. P, Chen, P. D., Liu, D. J., Kynast, R., Friebe, B., Velazhahan, R., Muthukrishnan, S. and Gill, B. S., Development of wheat scab symptoms is delayed in transgenic wheat plants that conStitutively express a rice thaumatin-like protein. Theor Appl Genet. 99, 755-760 (1999) https://doi.org/10.1007/s001220051294
  4. Chen, W. P., and Punja, Z. K., Transgenic herbic de-and disease-tolerant carrot (Daucus carota L.) plan.s obtained through Agrobacterium-mediated transformation Plant Cell Rept. 20, 929-935 (2002) https://doi.org/10.1007/s00299-001-0419-7
  5. Chilton, M. D., Currier, T. C., Farrand, S. K., Bendich, A. J., Gordon, M. P., and Nester, E. W., Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detectable in crown gall tumours. Proc Natl Acad Sci USA. 71, 3672-3676 (1974) https://doi.org/10.1073/pnas.71.9.3672
  6. Christensen, A. H. and Quail, P. H., Ubiquitirn promoterbased vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res. 5, 213-218 (1996) https://doi.org/10.1007/BF01969712
  7. Christensen, A. H., sharrock, R. A., and Quail, P. H., Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Mol Biol. 18, 675-689 (1992) https://doi.org/10.1007/BF00020010
  8. Datta, K., Velazhahan, R., Oliva, N., Ona, I., Mew, I., Khush, G. S., Muthukrishnan, S., and Datta, S. K., Over-expression of the cloned rice thaumatin-like protein (PR-5) gene in transgenic rice plants enhances environmental friendly resistance to Rhizoctonia solani causing sheath blight disease. Theor Appl Genet. 98, 1138-1145 (1999) https://doi.org/10.1007/s001220051178
  9. Fang, G., Hammar, S., and Grumet, R., A quick and inexpensive method for removing polysaccharides from plant genomic DNA BioTechniques 13, 52, 54, 56 (1992)
  10. Fukuoka, H., Ogawa, T., Matsuoka, M., Ohkawa, Y, and Yano, H., Direct gene delivery into isolated microspores of rapeseed (Brassica napus L.) and the production of fertile transgenic plants. Plant Cell Rept. 17, 323-328 (1998) https://doi.org/10.1007/s002990050401
  11. Gritz, L. and Davies, J., Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene. 25, 179-188 (1983) https://doi.org/10.1016/0378-1119(83)90223-8
  12. Henke, W., Herdel, K., Jung, K., schnorr, D., and Leoning, S. A, Betaine improves the PCR amplification cf GC-rich DNA sequences. Nucleic Acids Res. 25, 3967-3958 (1997) https://doi.org/10.1093/nar/25.19.3957
  13. Klimyuk, V. I., Carroll, B. J., Thomas, C. M., and Jones, J. D. G., Alkali treatment for rapid preparation of plant material for reliable PCR analysis. Plant J. 3, 493-494 (1993) https://doi.org/10.1046/j.1365-313X.1993.t01-26-00999.x
  14. Murashige, T. and Skoog, F., A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant. 15, 473-497 (1962) https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  15. Odell, J. T., Nagy, F., and Chua, N. H., Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313, 810-812 (1985) https://doi.org/10.1038/313810a0
  16. Proctor, J. T.A. and Bailey, W. G., Ginseng: industry, botany, and culture. Hort Rev. 9, 187-236 (1987)
  17. Punja, Z. K., Fungal pathogens of American ginseng (Panax quinquefolium) in British Columbia. Can J Plant Pathol. 19, 301-306 (1997) https://doi.org/10.1080/07060669709500528
  18. Reeleder, R. D. and Brammall, R. A, Pathogenicity of Pythium species, Cylindrocarpon destructans, and Rhizoctonia solani to ginseng seedlings in Ontario. Can J Plant Pathol. 16, 311-316 (1994) https://doi.org/10.1080/07060669409500736
  19. Riede, C. R. and Anderson, J. A, Linkage of RFLP markers to an aluminum tolerance gene in wheat. Crop Sci. 36, 905-909 (1996) https://doi.org/10.2135/cropsci1996.0011183X0036000400015x
  20. Takimoto, I., Christensen, A. H., and Quail, P. H., Non-systerric expression of a stress-responsive maize polyubiquitin gene (ubi-1) in transgenic rice plants. Plant Mol. Biol. 141, 51-58 (1994)
  21. Thompson, C. J., Movva, N. R., Tizard, R., Crameri, R., Davies, J. E., Lauwereys, M., and Botterman, J., Characterization of the herbicide-resistance gene bar from streptomyces hygroscopicus. EMBO J. 6, 2519-2523 (1987)
  22. Tingay, S., McElroy, D., Kalla, R., Fieg, S., Wang, M., Thornton, S., and Brettell, R., Agrobacterium tumefaciensmediated barley transformation. Plant J. 11, 1369-1376 (1997) https://doi.org/10.1046/j.1365-313X.1997.11061369.x
  23. Tirajoh, A, Kyung, T. S., and Punja, Z. K., somatic embryogenesis and plantlet regeneration in American ginseng (Panax quinquefolium L.). In Vitro Cell Dev Biol Plant. 34, 203-211 (1998) https://doi.org/10.1007/BF02822709
  24. Velazhahan, R., Chen-Cole, K., Anuratha, C. S., and Muthukri-shnan, S., Induction of thaumatin-like proteins (TLPs) in Rhizoctonia solani-infected rice and characterization of two new cDNA clones. Physiol Plant. 102, 21-28 (1998) https://doi.org/10.1034/j.1399-3054.1998.1020104.x
  25. White, J., Chang, S. Y. P., Bibb, M. J., and Bibb, M. J., A cassette containing the bar gene of streptomyces hygroscopicus: a selectable marker for plant transformation. Nucleic Acids Res. 18, 1062 (1990) https://doi.org/10.1093/nar/18.4.1062

Cited by

  1. vol.95, pp.12, 2005, https://doi.org/10.1094/PHYTO-95-1381