The effect of NbRbohB on Bamboo mosaic virus replication in Nicotiana benthamiana
| Autor: | Chia-Hsin Hsueh, 薛家欣 |
|---|---|
| Rok vydání: | 2016 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 104 Respiratory burst oxidase homologs (Rboh), alias NADPH oxidase, plays a critical role in reactive oxygen species (ROS) generation. In Nicotiana benthamiana, there are two kinds of Rboh, NbRbohA and NbRbohB. NbRbohB, induced by specific pathogen signals, participates in mechanisms for plant disease resistance. Bamboo mosaic virus (BaMV), a member of the Potexvirus genus, contains a single-stranded positive-sense RNA genome. A previous proteomic approach identified a potential interaction between NbRbohB and BaMV replicase. To explore the relationship between NbRbohB and BaMV replication, NbRbohB was down-regulated by the Tobacco rattle virus (TRV)-induced gene silencing method and the silencing plant was subsequently inoculated with the recombinant BaMV virion that carries green fluorescent protein gene (GFP) in this study. Western blot analysis, revealed that the accumulations of GFP and BaMV CP were reduced significantly in NbRbohB-silenced plants. Northern blot and RT-PCR also demonstrated that the accumulation of BaMV genomic RNA was decreased when NbRbohB was silenced. The same VIGS method was used to analyze the effects of NbRbohB on Foxtail mosaic virus (FoMV) and Potato virus X (PVX) replication. Silencing NbRbohB reduced the accumulation of FoMV CP but not PVX CP. According to public EST databases, the full-length cDNAs of NbRbohB1 and NbRbohB2 were acquired, and they were inserted into the expression vector pBI221. Overexpression of NbRbohB1 in BaMV-infected protoplasts significantly increased the accumulation of BaMV CP. In conclusion, NbRbohB1 plays a significant role in promoting BaMV and FoMV replication. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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