Similar restrictions applied to an approach where a transcription factor was inserted into the genome of the tobacco etch virus that induced the accumulation of colored anthocyanins in infected tissue. thaliana observation in roots is not possible. However, observation is only possible at the macroscopic scale and restricted to photosynthetic tissue and to A. This allows easy visualization of the otherwise symptomless viral infection by bleaching the infected tissue. For example, the recombinant turnip yellows virus (TuYV) TuM1s81 contains a hairpin insertion for silencing the Arabidopsis thaliana CHLI1 gene. Each marker has its advantages and inconveniences. Most approaches rely on inserting a marker into the viral genome that can be followed easily in infected organisms. Taken together, our results show that NanoLuc luciferase can be used to monitor infection with TuYV.įollowing virus infection in real-time and in a non-destructive manner is desirable for various reasons, and different techniques have been developed or adapted for this. Bioluminescence imaging of these plants after agro-infiltration with TuYV-N65-NL allowed the detection of the virus in systemically infected leaves.
benthamiana plants expressing the C-terminal domain of split NanoLuc. For the detection of split NanoLuc, we constructed transgenic N. Bioluminescence imaging detected TuYV-NL in inoculated and systemically infected leaves.
RT-PCR analysis indicated that the recombinant sequences were stable in N. perfoliata but reduced systemic infection and lower accumulation in A. ELISA confirmed systemic infection and similar accumulation of the recombinant viruses in N. Wild-type and recombinant viruses were agro-infiltrated in Nicotiana benthamiana, Montia perfoliata, and Arabidopsis thaliana. 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). We used the NanoLuc luciferase bioluminescent reporter system to detect turnip yellows virus (TuYV) in infected plants.
0 kommentar(er)
