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. 2021 Nov 26;12:733129. doi: 10.3389/fpls.2021.733129

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Copyright © 2021 Sarkar and Roy-Barman.

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) and the copyright owner(s) 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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Systemic silencing activity of dsRNA and its accumulation in the local (sprayed) and distal (unsprayed) leaves. (A) Fluorescence microscopy of wounded M. oryzaeGFP-inoculated leaves that were either sprayed with either TE (mock) or dsGFP in local and distal regions. Mean fluorescence intensity of GFP from approximately five plants (10 lesions) of each set was monitored 4 and 7 dpt. (B) Northern blot profile for GFP-specific dsRNA. The longevity and accumulation of RNA signals were assessed 3, 4, 5, and 7 dpt on locally and distally sprayed leaves. The total RNA isolated from unsprayed (US) leaves was run as negative control. In vitro-synthesized and purified dsGFP was used as the positive control (PC), and rice 25S rRNA was kept as the loading control. Sense GFP probe with the same sequence as the parent dsGFP and PCR-purified rice 25S rRNA (primers listed in Supplementary Table 2) were used for hybridization.