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Silencing of NbSKP1s enhances CLCuMuV DNA accumulation and results in typical disease symptoms.

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posted on 2016-06-17, 03:30 authored by Qi Jia, Na Liu, Ke Xie, Yanwan Dai, Shaojie Han, Xijuan Zhao, Lichao Qian, Yunjing Wang, Jinping Zhao, Rena Gorovits, Daoxin Xie, Yiguo Hong, Yule Liu

(A1, A2 and A3) Six- to seven-week-old N. benthamiana plants were agroinoculated with CLCuMuV (CA) and βM2-SKP1F1 (A1), βM2-SKP1F2 (A2), βM2-SKP1F3 (A3) or βM2-βC1F (as the control). (B1, B2 and B3) Silencing of NbSKP1s enhanced CLCuMuV DNA accumulation. Each group contained 7 plants. At 14 dpi, total DNA was extracted from each plant respectively and subjected to quantitative real-time PCR (means±SEM, n = 7) to quantify viral DNA accumulation. The internal reference method was used to calculate the relative amount of viral DNA. (C1, C2 and C3) Real-time RT-PCR confirmed silencing of NbSKP1s. Total RNA was extracted from each plant respectively and subjected to quantitative RT-PCR (means±SEM, n = 4) to quantify NbSKP1s mRNA level. Actin was used as the internal reference. The raw data of (B1–B3) and (C1–C3) were analysed by two-sample t-test to show the significance level at 0.05 (*) and 0.01 (**). These experiments were repeated at least twice. (D1, D2 and D3) 50% plants infected with CA+βM2-SKP1F1 (D1), 50% plants infected with CA+βM2-SKP1F2 (D2) and 100% plants infected with CA+βM2-SKP1F3 (D3) show severe symptoms at 21 dpi. (E1, E2, E3 and E4) Apical leaves of plants infected with CA+βM2-βC1F (E1), CA+βM2-SKP1F1(E2), CA+βM2-SKP1F2 (E3) and CA+βM2-SKP1F3 (E4) at 21 dpi.

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