GUS expression of the TNV-A^C recombinants.

<p>(A) Schematic illustration of wtTNV-A^C (pMTC 27), pTGUS-I and the recombinant derivatives pTGUS-II, pTGUS-I 17, pTGUS-I 81 and pTGUS-I 294. TNV ORFs are shown as gray boxes, the GUS gene as a white box, the 5′ proximal ends of the CP gene (nt 2612–2629 or nt 2612–2628) are illustrated as a box with diagonal lines, and the duplicate sequences upstream of the CP gene are represented by dotted boxes. The numbers identify the locations of the inserted nucleotide positions in TNV gRNA. (B) Inoculated <i>C. amaranticolor</i> leaf phenotypes were photographed at 4 dpi, and viral RNAs used for inoculations are shown above each photo (Top panel). The results of GUS histochemical staining are shown without magnification (Middle panel) or under a light microscope at 40X magnification (Bottom panel). The inoculated leaves were stained with 1 mM X-Gluc in 50 mM NaH₂PO₄ (pH 7.0) and incubated at 37°C in darkness overnight. (C) Viral RNA accumulation was assessed by Northern blots with the same probe used in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057938#pone-0057938-g001" target="_blank">Fig. 1</a>. The migration of viral RNAs and intensities of the rRNA loading control species are as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057938#pone-0057938-g001" target="_blank">Fig. 1</a>.