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. 2018 Apr 24;10(5):216. doi: 10.3390/v10050216

Figure 4.

Figure 4

Effects of heterologous CP subgenomic controller elements (CEs) on LIYV infection and GFP expression in Hc-Pro Nicotiana benthamiana plants. (A) Phenotypes of N. benthamiana plants inoculated with buffer (Mock), LIYV-WT, LIYV-L2CGFP (LIYV CP CE), LIYV-CCGFP (CYSDV CP CE), and LIYV-BCGFP (BYV CP CE) at 3 wpi. Symptoms on upper leaves (white light) and GFP fluorescence visualized under UV light (lower panels). Dotted lines indicate enlarged areas. (B) Detection of viral infection and insertion integrity by RT-PCR with total RNA extracted from upper non-inoculated leaves. Two primer sets were used to amplify the sequence of LIYV CP (CP, 530 bp) and the sequence flanking the GFP cassette (Insert, 1187 bp), LIYV-WT without the insert was amplified as a control (317 bp). (C) Immunoblot analysis of LIYV CP and GFP accumulation with total protein extracts from agroinoculated and upper non-inoculated leaves using LIYV CP and GFP specific antibodies. The Ponceau S stained rubisco large subunit serves as a loading control.