The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Transformation of Fuji Apple Plant Harboring the Coat Protein Gene of Cucumber mosaic virus

  • Lee, C.H. (Division of Bioscience and Technology, Korea University) ;
  • Hyung, N.I. (Department of Horticultural Science, Sangmyung University) ;
  • Lee, G.P. (Plant Virus GenBank, College of Natural Science, Seoul Women’s University) ;
  • Choi, J.Y. (Department of Horticultural Science, Sangmyung University) ;
  • Kim, C.S. (Division of Bioscience and Technology, Korea University) ;
  • Choi, S.H. (Plant Virus GenBank, College of Natural Science, Seoul Women’s University) ;
  • Jang, I.O. (Plant Virus GenBank, College of Natural Science, Seoul Women’s University) ;
  • Han, D.H. (Division of Bioscience and Technology, Korea University) ;
  • Ryu, K.H.
  • Published : 2003.06.01
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Abstract

Transformation of Fuji apple (Malus domestica 'Fuji') was performed using Agrobacterium tumefaciens harboring a coat protein (CP) gene of Cucumber mosaic virus (CMV). A plasmid DNA containing the virus CP and NPT II genes was introduced into the loaves of apple by th e Agrobacterium - mediated transformation procedure. Regenerated transformants of the apple were obtained by kanamycin resistance conferred by the introduced NPT II gene. PCR analysis showed that 3 out of 20 putatively selected R0 plant lines contain the CMV-CP gene. Nine putative transgenic lines out of 20 lines were investigated with the PCR analysis; 5 regenerants produced a 450 bp DNA band and 3 regenerants showed a 671 bp DNA band for the NPT II and CMV-CP genes, respectively. Southern hybyidization results demonstrate the successful integration of the CMV-CP gene into the genome of the apple. This is the first report on the generation of useful vius resistance source of transgenic apple for molecular breeding program.

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References

  1. De Bondt, A., Eggermont, K., Penninckx,I., Goderis, I., Vander-leyden, J. and Broekaert, W. F. 1996. Agrobacterium-mediated transformation of apple (Malus xdomestica Borkh): An assessment of factors affecting regeneration of transgenic plants. Plant Cell Rep. 15:549-554 https://doi.org/10.1007/BF00232992
  2. James, D. J., Passey, A. J., Barbara, D. J. and Bevan, M. 1989. Genetic transformation of apple (Malus pumila Mill.) using disarmed Ti-binary vector. Plant Cell Rep. 7:658-661
  3. Kim, C. S., Lee, C. H., Shin, J. S., Jung, Y. S. and Hyung N. I. 1997.A simple and rapid method for isolation of DNA from woody plant using PVP. Nucleic Acids Res. 25:1085-1086 https://doi.org/10.1093/nar/25.5.1085
  4. Lee, C. H., Hyung, N. I. and Kim, S. B. 1995. Foreign gene transfer using electroporation and transient expression in apple. Acta Hort. 392:179-185
  5. Lee, G. P., Ryu, K H., Kim, H. R., Kim, C. S., Lee, D. W., Kim, J. S., Park, M. H., Noh, Y. M., Choi, S. H., Han, D. H. and Lee, C. H. 2002. Cloning and phylogeneticcharacterization of coat protein genes of two isolates of Apple mosaic virus from 'Fuji' apple. Plant Pathol J. 18:259-265 https://doi.org/10.5423/PPJ.2002.18.5.259
  6. Maheswaran, G., Welander, M., Hutchinson, J. F., Graham, M. W. and Richard, D. 1992. Transformation of apple rootstock M26 with Agrobacterium tumefaciens. J. Plant Physiol. 139:560-568 https://doi.org/10.1016/S0176-1617(11)80370-6
  7. Martin, G. C, Miller, A. N., Castle, L. A., Morris, J. W., Morris, R. O. and Dandekar, A. M. 1990. Feasibility studies using - glucuronidase as a gene fusion marker in apple, peach and radish. J. Amer. Soc. Hart. Sci. 115:686-691
  8. Maximova, S. N., Dandekar, A. M. and Guiltinan, M. J. 1998. Investigation of Agrobacterium-mediated transformation of apple using green fluorescent protein: high transient expression and low stable transformation suggest that factors other than T-DNA transfer are rate-limiting. Plant Mol. BioI. 37:549-559 https://doi.org/10.1023/A:1006041313209
  9. Moore G. A, Jacono, C. C, Neidigh, J. L., Lawrence, S. D. and Cline, K. 1992. Agrobacterium-mediate transformation of citrus stem segments and regenerationof transgenic plants. Plant Cell Rep. 11 :238-242
  10. Norelli, J. L. and Aldwinckle, H. S. 1993.The role of aminoglycoside antibiotics in the regeneration and selection of neomycin phosphotransferase-transgenic apple trees. J. Am. Soc. Hart. Sci. 118:311-316
  11. Norelli, J. L., Adwinckle, H. S., Destefano-Beltran, L. and Jaynes, J. M. 1994. Transgenic Mailing 2 apple expressing the attacin E gene has increased resistance to Erwinia amylovora. Euphytica 77:123-128 https://doi.org/10.1007/BF02551474
  12. Ryu, K. H. and Park, W. M. 1995. Construction of plant expression vector for coat protein gene of cucumber mosaic virus As strain for plant transformation. Korean J. Plant Pathol. 11 :66-72
  13. Sambrook, J., Fritsch, E. F. and Maniatis, T. 1989. Molecular cloning: A laboratory manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
  14. Song, K. J., Ahn, S. Y., Hwang, J. H., Shin, Y. D., Park, S. W. and An, G. 2000. Agrobacterium-mediated transformation of McIntosh Wijick apple. J. Kor. Soc. Hort. Sci. 41:541-544
  15. Sriskandarjah, S., Goodwin, P. B. and Speirs, J. 1994. Genetic transformation of apple scion cultivar Delicious via Agrobacterium tumefaciens. Plant Cell Tissue Organ Cult. 36:317-329 https://doi.org/10.1007/BF00046089
  16. Yao, J., Golden, D., Atkinson, R., Richardson, K. and Morris, B. 1995. Regeneration of transgenic plants from the commercial apple cultivar Royal Gala. Plant Cell Rep. 14:407-412