Figure 1.

Northern blot analysis of the accumulation of alfalfa mosaic virus (AMV) RNA carrying citrus leprosis virus C (CiLV-C) and citrus leprosis virus C2 (CiLV-C2) movement proteins (MPs) in transgenic Nicotiana tabacum (P12) protoplasts. (A) The schematic representation shows the pAL3NcoP3 AMV RNA 3 construct [36] and its derivatives carrying the heterologous MPs fused to the C-terminal 44 residues of the AMV MP (A44), used in this assay. The open reading frames, represented by large boxes, correspond to the MP, and the coat protein (CP). The short box corresponds to the A44, meanwhile, arrow represents subgenomic promoter. (B) Northern blot showing accumulation of AMV RNAs in P12 protoplasts at 16 h post-inoculation, using a dig-riboprobe complementary to the 3′ UTR of the AMV RNA 3. Transcripts correspond to the AMV RNA 3 derivative carrying the CiLV-C MP, CiLV-C2 MP, and AMV MP (positive control) proteins. The localization of RNA 3 and subgenomic RNAs (sgRNA 4) are indicated. The graph represents the total accumulation of AMV RNA, carrying the constructs AMV MP, CiLV-C MP, and CiLV-C2 MP from the average of two independent experiments. The bands were quantified using the ImageJ version 2.0cr software with the ISAC plugin and error bars represent standard deviation. Statistical analysis was performed using Student’s t-test and one-way ANOVA. p values less than 0.05 (typically ≤ 0.05) are statistically significant. p-values were obtained from pairwise comparisons between treatments or among the three groups.