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Establishment of an Agrobacterium-mediated Inoculation System for Cucumber Green Mottle Mosaic Virus

  • Kang, Minji (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • Seo, Jang-Kyun (Crop Protection Division, National Academy of Agricultural Science) ;
  • Song, Dami (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University) ;
  • Choi, Hong-Soo (Crop Protection Division, National Academy of Agricultural Science) ;
  • Kim, Kook-Hyung (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University)
  • Received : 2015.06.26
  • Accepted : 2015.07.23
  • Published : 2015.12.01

Abstract

The infectious full-length cDNA clones of Cucumber green mottle mosaic virus (CGMMV) isolates KW and KOM, which were isolated from watermelon and oriental melon, respectively, were constructed under the control of the cauliflower mosaic virus 35S promoter. We successfully inoculated Nicotiana benthamiana with the cloned CGMMV isolates KW and KOM by Agrobacterium-mediated infiltration. Virulence and symptomatic characteristics of the cloned CGMMV isolates KW and KOM were tested on several indicator plants. No obvious differences between two cloned isolates in disease development were observed on the tested indicator plants. We also determined full genome sequences of the cloned CGMMV isolates KW and KOM. Sequence comparison revealed that only four amino acids (at positions 228, 699, 1212, and 1238 of the replicase protein region) differ between the cloned isolates KW and KOM. A previous study reported that the isolate KOM could not infect Chenopodium amaranticolor, but the cloned KOM induced chlorotic spots on the inoculated leaves. When compared with the previously reported sequence of the original KOM isolate, the cloned KOM contained one amino acid mutation (Ala to Thr) at position 228 of the replicase protein, suggesting that this mutation might be responsible for induction of chlorotic spots on the inoculated leaves of C. amaranticolor.

Keywords

References

  1. Dawson, W. O., Lewandowski, D. J., Hilf, M. E., Bubrick, P., Raffo, A. J., Shaw, J. J., Grantham, G. L. and Desjardins, P. R. 1989. A tobacco mosaic virus-hybrid expresses and loses an added gene. Virology 172:285-292. https://doi.org/10.1016/0042-6822(89)90130-X
  2. Domingo, E., Martinez-Salas, E., Sobrino, F., de la Torre, J. C., Portela, A., Ortin, J., Lopez-Galindez, C., Perez-Brena, P., Villanueva, N., Najera, R. et al. 1985. The quasispecies (extremely heterogeneous) nature of viral RNA genome populations: biological relevance--a review. Gene 40:1-8.
  3. Hajimorad, M. R., Eggenberger, A. L. and Hill, J. H. 2003. Evolution of Soybean mosaic virus-G7 molecularly cloned genome in Rsv1-genotype soybean results in emergence of a mutant capable of evading Rsv1-mediated recognition. Virology 314:497-509. https://doi.org/10.1016/S0042-6822(03)00456-2
  4. Hebrard, E., Pinel-Galzi, A., Bersoult, A., Sire, C. and Fargette, D. 2006. Emergence of a resistance-breaking isolate of Rice yellow mottle virus during serial inoculations is due to a single substitution in the genome-linked viral protein VPg. J. Gen. Virol. 87:1369-1373. https://doi.org/10.1099/vir.0.81659-0
  5. Kim, S. M., Lee, J. M., Yim, K. O., Oh, M. H., Park, J. W. and Kim, K.-H. 2003. Nucleotide sequences of two Korean isolates of Cucumber green mottle mosaic virus. Mol. Cells 16:407-412.
  6. Lee, K. Y. 1996. Current occurrence and control of CGMMV 'Konjak' disease. Plant Dis. Agric. 2:38-39.
  7. Lee, K. Y., Lee, B. C. and Park, H. C. 1990. Occurrence of cucumber green mottle mosaic virus disease of watermelon in Korea. Kor. J. Plant Pathol. 6:250-255.
  8. Park, S.-H. and Kim, K.-H. 2006. Agroinfilteration-based Potato virus X replicons to dissect the requirements of viral infection. Plant Pathol. J. 22:386-390. https://doi.org/10.5423/PPJ.2006.22.4.386
  9. Ruiz, M. T., Voinnet, O. and Baulcombe, D. C. 1998. Initiation and maintenance of virus-induced gene silencing. Plant Cell 10:937-946. https://doi.org/10.1105/tpc.10.6.937
  10. Seo, J.-K., Lee, S.-H. and Kim, K.-H. 2009a. Strain-specific cylindrical inclusion protein of soybean mosaic virus elicits extreme resistance and a lethal systemic hypersensitive response in two resistant soybean cultivars. Mol. Plant-Microbe Interact. 22:1151-1159. https://doi.org/10.1094/MPMI-22-9-1151
  11. Seo, J.-K., Lee, H. G. and Kim, K.-H. 2009b. Systemic gene delivery into soybean by simple rub-inoculation with plasmid DNA of a Soybean mosaic virus-based vector. Arch. Virol. 154:87-99. https://doi.org/10.1007/s00705-008-0286-4
  12. Seo, J.-K., Sohn, S.-H. and Kim, K.-H. 2011. A single amino acid change in HC-Pro of soybean mosaic virus alters symptom expression in a soybean cultivar carrying Rsv1 and Rsv3. Arch. Virol. 156:135-141. https://doi.org/10.1007/s00705-010-0829-3
  13. Ugaki, M., Tomiyama, M., Kakutani, T., Hidaka, S., Kiguchi, T., Nagata, R., Sato, T., Motoyoshi, F. and Nishiguchi, M. 1991. The complete nucleotide sequence of cucumber green mottle mosaic virus (SH strain) genomic RNA. J. Gen. Virol. 72:1487-1495. https://doi.org/10.1099/0022-1317-72-7-1487
  14. Zhong, M., Zhao, X., Liu, Y., Wang, Y. and Cao, K. 2015. Completion sequence and cloning of the infectious cDNA of a chb isolate of cucumber green mottle mosaic virus. Acta Virol. 59:49-56. https://doi.org/10.4149/av_2015_01_49

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