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Volume 5, Issue 5, Pages 1151-1153 (September 2012)
SGT1 Is Induced by the Potato virus X TGBp3 and Enhances Virus Accumulation in Nicotiana benthamiana Chang-Ming Ye, Vicky Kelly, Mark Payton, Martin B. Dickman, Jeanmarie Verchot Molecular Plant Volume 5, Issue 5, Pages (September 2012) DOI: /mp/sss026 Copyright © 2012 The Authors. All rights reserved. Terms and Conditions
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Figure 1 Plot represents the increasing percent of nontransgenic (♦) and SGT662 (▪) plants that were systemically infected with PVX-GFP between 3 and 7 dpi. Each day the percent of infected SGT662 plants is higher than nontransgenic. (A–C) qRT–PCR detecting NbSGT1 transcripts in N. benthamiana plants. The comparative CT method employs the formula 2 –▵▵CT to represent the RNA fold accumulation. The endogenous control (18S RNA) and calibrator (healthy sample template) were used to calculate the ddCT value. Validation reactions determined the efficiencies of the target and control amplifications. (A) Boxplots representing data obtained from 20 PVX–GFP-infected leaves. (B) Bars represent the average (n = 3) relative levels of NbSGT1 at 2 and 5 dpi in leaves that were infiltrated with buffer (M); A. tumefaciens only (Agro) or containing CaMV35S driven PVX–GFP; mycRep (Rep); TGBp1; mycTGBp2; TGBp3His; or CP. (C) Bars represent NbSGT1 induction (n = 3) at 2 d following leaf infiltrations with buffer (mock) or various dilutions of Agro containing CaMV35S–TGBp3 or Nos–TGBp3 fusions. ANOVA verified TGBp3-induced higher levels of host transcripts (p < 0.05). (D) Ethidium bromide-stained gel showing semi-quantitative RT–PCR products obtained using RNA extract from healthy, TRV, or TRV–SGT1-treated leaves and primers detecting NbSGT1 or actin. The sizes in base pairs (bp) of the DNA ladder (L) are indicated. The numbers of PCR cycles are indicated. (E) Fourteen days after pre-treatment with buffer (0), TRV and TRV–NbSGT1 (indicated above the lanes) plants were inoculated with PVX–GFP. Immunoblot detecting PVX CP in systemic leaf extracts a 5-dpi Coomassie blue-stained gel below the immunoblot shows equal sample loading. The percentage values at the bottom indicate the proportion of plants that were systemically infected at 5 dpi. (F) PVX–GFP-infected SGT662 and nontransgenic plants at 5 dpi. (G) Immunoblot detecting PVX CP in systemic leaves at 5 dpi in SGT662 and nontransgenic plants. (H) Plot shows the percentage of plants that showed systemic PVX–GFP accumulation between 3 and 7 dpi. Molecular Plant 2012 5, DOI: ( /mp/sss026) Copyright © 2012 The Authors. All rights reserved. Terms and Conditions
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