Abstract
Cucumber green mottle mosaic virus (CGMMV) is a major limiting factor in the production of melon plants worldwide. For effective control of this virus using the transgenic approach, the direct repeat of the movement protein gene of CGMMV was used for transforming melon plants by Agrobacterium tumefaciens. PCR and Southern blot analyses of T3 confirmed that they carried the transgene. Northern blot analysis with total RNA showed that transgene transcript RNA as well as siRNA was observed in all plants tested. Separate leaves or individual plants were inoculated with CGMMV and subjected to ELISA and RNA blot analysis using the coat protein gene probe of the virus. Compared to nontransgenic control, these plants were shown to have high virus resistance. Furthermore, cytosine of the transgene DNA in the plants was methylated. Thus, these results reveal that the transgenic lines were highly resistant to the virus through RNA silencing.
Key message High virus resistance was obtained in transgenic melon plants with direct repeat of movement protein gene of Cucumber green mottle mosaic tobamovirus through RNA silencing.
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Acknowledgments
We are grateful to F. Savattini and Q. Xie at the National Institute of Agrobiological Sciences, Tsukuba, Japan, for their technical assistance. We thank D. Murphy for English corrections. This study was supported partially by grants from the Ministry of Agriculture, Forestry and Fisheries of Japan, the Ministry of Education, Culture, Sports, Science and Technology of Japan, and Grants in Aid for Scientific Research 00465001 to MN and 21380032 to KK.
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Communicated by H. Ebinuma.
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Emran, A.M., Tabei, Y., Kobayashi, K. et al. Molecular analysis of transgenic melon plants showing virus resistance conferred by direct repeat of movement gene of Cucumber green mottle mosaic virus . Plant Cell Rep 31, 1371–1377 (2012). https://doi.org/10.1007/s00299-012-1237-9
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DOI: https://doi.org/10.1007/s00299-012-1237-9