Silencing potential of viral derived RNAi constructs in Tomato leaf curl virus-AC4 gene suppression in tomato
Autor: | Vikas Koundal, Anil Kumar Mishra, Suhas Ramesh, Peter Palukaitis, Shelly Praveen |
---|---|
Rok vydání: | 2009 |
Předmět: |
Gene Expression Regulation
Viral Small interfering RNA Genes Viral viruses Trans-acting siRNA Efficiency Biology Models Biological Plant Viruses Transformation Genetic Solanum lycopersicum Gene Expression Regulation Plant RNA interference Transcription (biology) Sense (molecular biology) Genetics Gene silencing Gene Silencing Cloning Molecular RNA Small Interfering Gene Plant Diseases fungi Plants Genetically Modified Immunity Innate RNA silencing Animal Science and Zoology Agronomy and Crop Science Plasmids Biotechnology |
Zdroj: | Transgenic Research. 19:45-55 |
ISSN: | 1573-9368 0962-8819 |
DOI: | 10.1007/s11248-009-9291-y |
Popis: | We investigated viral gene suppression in an infected tomato, by transforming it with RNA inhibition (RNAi) constructs derived from same viral gene. To develop RNAi constructs, conserved sequences ranging from 21 to 200 nt of the viral target AC4 gene of various viruses causing the tomato leaf curl disease were chosen. The double-stranded (ds)RNA producing constructs carry the sense and antisense portions of these sequences and are separated by different introns behind a constitutive promoter. We compared the levels of suppression of the viral target gene by transforming four different RNAi constructs with varied arm length of dsRNA. Gene silencing levels of the viral target gene were found to be directly proportional to the arm length of the dsRNA. We observed that dsRNA derived from longer arm-length constructs generating a pool of siRNAs that were more effective in targeting gene silencing. After transformation, one of the RNAi construct having a 21 nt arm-length produced aberrant phenotypes. These phenotypic anomalies may be due to unintended ('off-target') host transcript silencing. The unintended host transcript silencing showed modest reversion in the presence of the viral target gene. The findings presented here suggest that the arm length of dsRNA capable of producing a pool of diced siRNAs is more efficient in gene silencing, the effect of off-targeting siRNA is minimized in a pool, and off-targeting silencing can be minimized in the presence of target gene. |
Databáze: | OpenAIRE |
Externí odkaz: |