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Characterization of resistance mechanism in transgenic Nicotiana benthamiana containing Turnip crinkle virus coat protein
- Source :
- Plant Cell Reports. 27:1731-1740
- Publication Year :
- 2008
- Publisher :
- Springer Science and Business Media LLC, 2008.
-
Abstract
- Two transgenic lines, of Nicotiana benthamiana expressing Turnip crinkle virus (TCV)-coat protein (CP) gene with contrasting phenotype, the highest (#3) and the lowest (#18) CP expressers, were selected and challenged with the homologous TCV. The former, the highest expresser, showed nearly five times more CP expression than the latter. Progenies of #3 and #18 lines showed 30 and 100% infection rates, respectively. The infected progenies of #3 line showed mild and delayed symptom with TCV. This is a coat protein-mediated resistance (CP-MR), and its resistance level is directly proportional to CP transgene expression. However, CP-MR of the transgenic plants was specific only for TCV but not for heterologous viruses. Newly growing leaves of those infected progenies of #3 line did not show any visible symptoms at 4-week post-inoculation (wpi) with TCV, suggesting a reversal from infection. This was confirmed by RT-PCR analysis with the disappearance of the target at 4 wpi. This is a case of RNA-mediated resistance, and a threshold level of transgene expression may be needed to achieve the silent state. To confirm the RNA silencing, we infiltrated Agrobacterium carrying TCV-CP into leaves of progenies of #3 and performed RT-PCR analysis. The results indicate that TCV-CP's suppressor activity against RNA silencing itself can be silenced by the homologous expression of TCV-CP in the transgenic plants. The transgenic plants containing TCV-CP seem to be a model system to study viral protection mediated by a combination of protein and RNA silencing.
- Subjects :
- Base Sequence
Transcription, Genetic
biology
Reverse Transcriptase Polymerase Chain Reaction
Agrobacterium
Turnip crinkle virus
Transgene
Carmovirus
Blotting, Western
Nicotiana benthamiana
RNA
Enzyme-Linked Immunosorbent Assay
Plant Science
General Medicine
Plants, Genetically Modified
biology.organism_classification
Virology
Tombusviridae
RNA silencing
Tobacco
Capsid Proteins
Agronomy and Crop Science
DNA Primers
Subjects
Details
- ISSN :
- 1432203X and 07217714
- Volume :
- 27
- Database :
- OpenAIRE
- Journal :
- Plant Cell Reports
- Accession number :
- edsair.doi.dedup.....94321e9791b36032ec03d49bb2071879