Functional Analysis of Cotton Leaf Curl Kokhran Virus/Cotton Leaf Curl Multan Betasatellite RNA Silencing Suppressors
Abstract
:1. Introduction
2. Materials and Methods
2.1. Generation of Constructs
2.2. Plants and Plant Transformation
2.3. Agro-Infiltration Patch Assay
2.4. Expression and Purification of Recombinant Proteins
2.5. Electrophoretic Mobility Shift Assay (EMSA)
2.6. SDS-PAGE
2.7. Western Blotting
3. Results
3.1. Suppression of S-PTGS
3.2. Suppression of IR-PTGS
3.3. Effects of Virus Proteins on Systemic Silencing
3.4. Interactions of Proteins with Nucleic Acids
3.5. Effects of Transgenic Expression of Protein in Plants
Inoculum | No. of plant systemically silenced/no. of plants inoculated | Silencing(%) |
---|---|---|
GFP+GUS | 27/49 | 55 C |
GFP+p19 | 2/27 | 7 A |
GFP+TrAP | 11/27 | 41 B |
GFP+C4 | 1/29 | 3 A |
GFP+V2 | 15/27 | 56 B |
GFP+ βC1 | 30/44 | 68 C |
IR-GFP+GUS | 28/48 | 58 C |
IR-GFP+p19 | 2/28 | 7 A |
IR-GFP+TrAP | 13/27 | 48 B |
IR-GFP++C4 | 2/30 | 6 A |
IR-GFP+V2 | 16/26 | 62 B |
IR-GFP+βC1 | 32/44 | 73 C |
4. Discussion
5. Conclusions
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Saeed, M.; Briddon, R.W.; Dalakouras, A.; Krczal, G.; Wassenegger, M. Functional Analysis of Cotton Leaf Curl Kokhran Virus/Cotton Leaf Curl Multan Betasatellite RNA Silencing Suppressors. Biology 2015, 4, 697-714. https://doi.org/10.3390/biology4040697
Saeed M, Briddon RW, Dalakouras A, Krczal G, Wassenegger M. Functional Analysis of Cotton Leaf Curl Kokhran Virus/Cotton Leaf Curl Multan Betasatellite RNA Silencing Suppressors. Biology. 2015; 4(4):697-714. https://doi.org/10.3390/biology4040697
Chicago/Turabian StyleSaeed, Muhammad, Rob W. Briddon, Athanasios Dalakouras, Gabi Krczal, and Michael Wassenegger. 2015. "Functional Analysis of Cotton Leaf Curl Kokhran Virus/Cotton Leaf Curl Multan Betasatellite RNA Silencing Suppressors" Biology 4, no. 4: 697-714. https://doi.org/10.3390/biology4040697