The role of post-Golgi transport pathways and sorting motifs in the plasmodesmal targeting of the movement protein (MP) of Ourmia melon virus (OuMV)

Natali Ozber; Paolo Margaria; Charles T. Anderson; Massimo Turina; Cristina Rosa

The effect of specific amino acid substitutions in MP on virus replication was measured by agroinfiltrating local leaves with pGC-RNA2 (MP) or mutants, together with pGC-RNA1 (RdRP), pGC-RNA3 (CP), and pBin61-GFP at an OD600 of 1.0. The GFP . CC-BY-NC-ND 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/724716 doi: bioRxiv preprint signal at the infiltrated sites was monitored at 36 hpi to confirm that agroinfiltrated areas were saturated. Two leaf spots from 3 leaves were pooled for each sample at 24 hpi and 48 hpi. The virus accumulation was measured by quantifying RNA1 by qPCR using primers listed in Table S1 . The qPCR was performed in duplicates using iTaq™ Universal SYBR® Green Supermix (BioRad) and a CFX96 (BioRad), following the manufacturer's instructions. The 2 -ΔΔCT method was used to asses fold changes in replication of wild-type and mutants. The data was normalized first using 18S ribosomal RNA, and then a wild-type control at 24 hpi showing the lowest expression. Four plants were used for qPCR analysis. Statistical analysis was performed using ANOVA.