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Pha13 is induced by CymMV and involved in virus accumulation.

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posted on 2018-09-13, 18:15 authored by Li Chang, Ho-Hsiung Chang, Jui-Che Chang, Hsiang-Chia Lu, Tan-Tung Wang, Duen-Wei Hsu, Yuh Tzean, An-Po Cheng, Yi-Shu Chiu, Hsin-Hung Yeh

(A) The CymMV accumulation level and expression level of Pha13 were analyzed by qRT-PCR in healthy (Mock) and CymMV-infected (Cy-infected) P. aphrodite. The RNA level in the CymMV was set to 1. Data represent mean ± SD; n = 3 biological replicates; *, P < 0.05, Student’s t-test compared to mock. (B and C) Transient silencing or overexpression of Pha13 in CymMV-infected plants. RNA level of Pha13 and CymMV were analyzed by qRT-PCR from leaves of CymMV-infected P. aphrodite infiltrated with agrobacterium carrying vector (Vector); hairpin RNA (hpRNA) vector to knockdown Pha13 (hpPha13; B); or overexpression vector of Pha13 (Pha13-oe; C). The RNA level of vector was set to 1. Data represent mean ± SD; n = 3 biological replicates; *, P < 0.05, Student’s t-test compared to vector. D, The protein level of Pha13 in WT and transgenic P. equestris (Pha13#27, #29 and #30) expressing Pha13, Pha13-oe (35S::Flag-Pha13), were analyzed by anti-Pha13 antibody. The anti-Tubulin antibody (Anti-Tub) was used as a loading control. (E) Three wild-type plants and three plants derived from the same transformed protocorm were used for analysis. RNA level of Pha13 and CymMV were analyzed by qRT-PCR from leaves of WT or transgenic P. equestris (Pha13#27, #29 and #30) inoculated with CymMV. The RNA level of WT was set to 1. Data represent mean ± SD; n = 3 biological replicates; *, P < 0.05, Student’s t-test compared to WT. PhaUbiquitin 10 was used as an internal control for normalization.

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