Rapid induction of transcriptional and post-transcriptional gene silencing using a novel Cucumber mosaic virus vector
-
- Otagaki Shungo
- Graduate School of Agriculture, Hokkaido University
-
- Arai Makoto
- Graduate School of Agriculture, Hokkaido University
-
- Takahashi Akiko
- Graduate School of Agriculture, Hokkaido University
-
- Goto Kazunori
- Graduate School of Agriculture, Hokkaido University
-
- Hong Jin-Sung
- Graduate School of Agriculture, Hokkaido University
-
- Masuta Chikara
- Graduate School of Agriculture, Hokkaido University
-
- Kanazawa Akira
- Graduate School of Agriculture, Hokkaido University
Search this article
Description
We developed a novel RNA virus vector based on the Cucumber mosaic virus (CMV), which is able to efficiently induce gene silencing in plants. We manipulated the RNA 2 of the CMV Y strain, whose genome consists of tripartite components, and introduced restriction sites for cloning a foreign sequence into the vector. To evaluate the vector (designated CMV2-A1) in terms of the ability to induce gene silencing, we cloned portions of the green fluorescent protein (GFP) cDNA or Cauliflower mosaic virus (CaMV) 35S promoter sequences into the vector and inoculated the infectious transcripts into Nicotiana benthamiana plants that express the GFP gene under the control of the CaMV 35S promoter. In both cases, a loss of GFP fluorescence accompanying a reduction in the level of GFP mRNA was induced. The short interfering RNAs (siRNAs) harboring the sequences inserted in the CMV2-A1 vector were detected in the silenced plants. When plants were infected with the virus containing the CaMV 35S promoter sequence, the CaMV 35S promoter sequence in the genomic DNA was heavily methylated. A reduction in the mRNA level of the GFP gene and loss of GFP fluorescence were induced as early as 6 and 12 days post-inoculation, respectively, earlier than the 20 days previously achieved with a Potato virus X vector. These results suggest that the CMV2-A1 vector is suitable for the rapid induction of both transcriptional and post-transcriptional gene silencing.
Journal
-
- Plant Biotechnology
-
Plant Biotechnology 23 (3), 259-265, 2006
Japanese Society for Plant Biotechnology
- Tweet
Keywords
Details 詳細情報について
-
- CRID
- 1390282679303638272
-
- NII Article ID
- 10021908671
-
- NII Book ID
- AA11250821
-
- COI
- 1:CAS:528:DC%2BD28XntVWhtL4%3D
-
- ISSN
- 13476114
- 13424580
- http://id.crossref.org/issn/13424580
-
- NDL BIB ID
- 7943819
-
- Text Lang
- en
-
- Data Source
-
- JaLC
- NDL
- Crossref
- CiNii Articles
-
- Abstract License Flag
- Disallowed