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. 2020 Mar 25;37(1):83–88. doi: 10.5511/plantbiotechnology.19.1227a

Figure 2. VIGS against the PDS gene in the spinach cultivar ‘All Right.’ (A) Photobleaching phenotype observed on the upper systemic leaves of spinach inoculated with L1A1-PDS2L3 at 10 dpi. The pictures of leaves of healthy plants and empty vector-infected plants are shown for comparison. (B) PDS expression levels in the upper non-inoculated leaves at 10 dpi. Real-time RT-PCRs were conducted using primer pairs as in Supplementary Table S1. The glyceraldehyde 3-phosphate (GAPDH) gene was used as a reference gene. Means (±SE) among the infected plants were analyzed for significant differences using Tukey’s multiple comparison test (* p<0.05); different letters above the bars indicate a significant difference among isolates. (C) CMV accumulation levels in upper non-inoculated leaves at 10 dpi. Real-time RT-PCR for the virus accumulation was conducted using primer pair of CMV-DET-5-340 and CMV-DET-3-340 (Supplementary Table S1). The CMV-DET-5-340 primer hybridizes to both CMV-L RNA3 (positions 1864–1884) and the subgenomic RNA from RNA3 designated RNA4, which is the mRNA for the coat protein. The CMV-DET-3-340 primer hybridizes to the 3′ end of RNA3 and RNA4. The size of the PCR product is 353 bp. The GAPDH gene was used as a reference (Supplementary Table S1). Statistical analysis was conducted by Student’s t-test (* p<0.05).

Figure 2. VIGS against the PDS gene in the spinach cultivar ‘All Right.’ (A) Photobleaching phenotype observed on the upper systemic leaves of spinach inoculated with L1A1-PDS2L3 at 10 dpi. The pictures of leaves of healthy plants and empty vector-infected plants are shown for comparison. (B) PDS expression levels in the upper non-inoculated leaves at 10 dpi. Real-time RT-PCRs were conducted using primer pairs as in Supplementary Table S1. The glyceraldehyde 3-phosphate (GAPDH) gene was used as a reference gene. Means (±SE) among the infected plants were analyzed for significant differences using Tukey’s multiple comparison test (* p<0.05); different letters above the bars indicate a significant difference among isolates. (C) CMV accumulation levels in upper non-inoculated leaves at 10 dpi. Real-time RT-PCR for the virus accumulation was conducted using primer pair of CMV-DET-5-340 and CMV-DET-3-340 (Supplementary Table S1). The CMV-DET-5-340 primer hybridizes to both CMV-L RNA3 (positions 1864–1884) and the subgenomic RNA from RNA3 designated RNA4, which is the mRNA for the coat protein. The CMV-DET-3-340 primer hybridizes to the 3′ end of RNA3 and RNA4. The size of the PCR product is 353 bp. The GAPDH gene was used as a reference (Supplementary Table S1). Statistical analysis was conducted by Student’s t-test (* p<0.05).