Abstract
Pattern-triggered immunity is the first line of defense against infection by pathogens such as bacteria and fungi in plants, and this mechanism remains poorly defined in plant viruses. Double-stranded RNA (dsRNA) is an intermediate in the replication of plant RNA viruses, and is considered to be a conserved structure of plant viruses similar to pathogen-associated molecular pattern. Whether dsRNA is the elicitor that activates plant immunity in response to virus infection remains obscure. In this method, we use the cDNA of turnip mosaic virus genome as the template to in vitro synthesis of viral dsRNA and examine whether viral dsRNA could activate plant immunity in Arabidopsis thaliana, including MAPK kinase cascade and reactive oxygen burst. In order to provide some references for researchers studying dsRNA in terms of research methodology and experimental methods, we use western blot to measure MAPK kinase cascade and luminol-based assay to measure ROS burst in Arabidopsis thaliana treated by viral dsRNA.
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Acknowledgments
This research is funded by grants from the National Natural Science Foundation of China (3207016) to GW, the China Agriculture Research System of MOF and MARA (CARS-24-C-04) to FY, and the K. C. Wong Magna Fund in Ningbo University.
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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Rui, P., Chen, J., Yan, F., Wu, G. (2024). Analysis of Plant Virus-Induced Immunity by Using Viral-Derived Double-Stranded RNA in Arabidopsis thaliana. In: Cheng, X., Wu, G. (eds) Double-Stranded RNA. Methods in Molecular Biology, vol 2771. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3702-9_14
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DOI: https://doi.org/10.1007/978-1-0716-3702-9_14
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