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We used the NanoLuc luciferase bioluminescent reporter system to detect turnip yellows virus (TuYV) in infected plants. For this, TuYV was genetically tagged by replacing the C-terminal part of the RT protein with full-length NanoLuc (TuYV-NL) or with the N-terminal domain of split NanoLuc (TuYV-N65-NL). Wild-type and recombinant viruses were agro-infiltrated in , , and . ELISA confirmed systemic infection and similar accumulation of the recombinant viruses in and but reduced systemic infection and lower accumulation in . RT-PCR analysis indicated that the recombinant sequences were stable in and but not in . Bioluminescence imaging detected TuYV-NL in inoculated and systemically infected leaves. For the detection of split NanoLuc, we constructed transgenic plants expressing the C-terminal domain of split NanoLuc. Bioluminescence imaging of these plants after agro-infiltration with TuYV-N65-NL allowed the detection of the virus in systemically infected leaves. Taken together, our results show that NanoLuc luciferase can be used to monitor infection with TuYV.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9692398 | PMC |
| http://dx.doi.org/10.3390/ijms232213685 | DOI Listing |
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