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. 2017 Sep 6;8:1689. doi: 10.3389/fmicb.2017.01689

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Copyright © 2017 Zhang, Liu, Liu, Cao, Massart and Wang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Agroinoculation infectivity assay of cDNA clone on N. benthamiana. (A) Construction of infectious cDNA clone. Whole viral genome was cloned using fragments A and B digested with StuI, NcoI and SalI and ligated with StuI-SalI digested pCB301 vector, 2 × 35S: promoter, RZ: ribozyme, NOS: teminator. (B) Confirmation of the constructs obtained by PCR, M5: DL 5 000 DNA marker, 1: pEASYT5-A, 2: pEASYT5-B, 3: pCB301-WLYaV. (C) Symptoms on infected N. benthamiana 4 weeks after infiltration of lower 3~4 leaves; upper leaves are systemically infected. Left: inoculated with pCB301 as negative control, no obvious symptoms; right: inoculated with pCB301-WLYaV, stunted plants have leaf yellowing. (D) Detection of WLYaV in N. benthamiana by RT-PCR 14 days post-inoculation, M2: DL 2 000 DNA marker, 17/24 plants were positive, NC: negative control (pCB301-inoculated N. benthamiana), PC: positive control (JN-U3), +: exhibiting yellowing symptom.