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. 2011 May 5;7(5):e1002021. doi: 10.1371/journal.ppat.1002021

Figure 4. Sequence complementarity between Y-sat and the host ChlI gene is important for the induction of yellowing.

Figure 4

A. The 22-nt complementary sequences of the ChlI genes of various plants and Y-sat or Y-sat mutants. The 22-nt complementary sequences in the ChlI genes (yellow region; YR) and in the Y-sat (satellite yellow region; SYR) are in bold face. Site-directed mutations were introduced in the SYR of Y-sat so that the generated Y-sat mutants (Y-sat mut-Tom, Y-sat mut-Ara1 and Y-sat mut-Ara2) have the 22-nt complementary sequence including G-U pairs to the host YR in ChlI of tomato, Arabidopsis ChlI1 and ChlI2, respectively. Introduced mutations of the Y-sat mutants are underlined. B. Yellow phenotype of pepper infected with CMV-Y (left) or CMV-Y and Y-sat (right) at 10 dpi. Pepper has the same YR sequence in the ChlI gene as that of tobacco. C. Yellow phenotype of tomato infected with CMV-Y and Y-sat mutant, Y-sat mut-Tom at 10 dpi. Left, tomato plant infected with CMV-Y and Y-sat mut-Tom; right, tomato plant infected with CMV-Y and Y-sat. D. Green mosaic on N. benthamiana infected with CMV-Y and Y-sat mut-Tom at 14 dpi. Left, N. benthamiana infected with CMV-Y and Y-sat mut-Tom; right, N. benthamiana infected with CMV-Y and Y-sat. E. Yellow phenotype of Arabidopsis infected with CMV-Y and Y-sat mut-Ara1 at 10 dpi. F. Green mosaic symptoms on N. benthamiana infected with CMV-Y and Y-sat mut-Ara1 at 14 dpi. Left, N. benthamiana infected with CMV-Y; middle, N. benthamiana infected with CMV-Y and Y-sat; right, N. benthamiana infected with CMV-Y and Y-sat mut-Ara1. G. The mRNA levels of ChlI in Y-sat mutant-infected N. benthamiana determined by quantitative real-time RT-PCR. RNAs were extracted from equivalent systemic leaves infected with CMV, CMV+Y-sat or CMV+Y-sat mutant, and equivalent leaves of healthy N. benthamiana. ChlI mRNA levels relative to the actin mRNA level are shown (mean ± SE; n = 3). H. SatRNAs accumulation in Y-sat- and Y-sat mut-Ara1-infected N. benthamiana. Total RNAs were extracted from the N. benthamiana leaves shown in panel F. An ethidium bromide-stained 1.2% agarose gel is shown. I. CMV accumulation at 14 dpi in systemic leaves of N. benthamiana inoculated with CMV-Y and Y-sat mut-Ara1, which was determined by conventional ELISA using antibodies raised against CMV CP. Samples are those shown in panel F.