Erratum

Erratum for the Research Article “Hierarchical action and inhibition of plant Dicer-like proteins in antiviral defense” by A. Deleris, J. Gallego-Bartolome, J. Bao, K. D. Kasschau, J. C. Carrington, O. Voinnet

Science  22 Jan 2016:
Vol. 351, Issue 6271,
DOI: 10.1126/science.aaf2336

In the Research Article “Hierarchical action and inhibition of plant Dicer-like proteins in antiviral defense,” the following mistakes were made by Olivier Voinnet, the corresponding author, during the final stages of figure mounting. The original lab book data provided to the editors of Science showed that these errors did not alter the data in any material way that could be construed to benefit the results and their conclusions. Olivier Voinnet takes the full responsibility for the mistakes. All the authors have approved the following corrections and wish to apologize deeply for the inconvenience caused.

Figure 1D depicts TRV-PDS infections of various Arabidopsis silencing mutants. The published Panel 1D was inappropriately mounted with the wrong ribosomal RNA loading control and this has created confusion in its final layout, in which the left-hand side of the blot, devoid of RNA, was wrongly set as a mock (i.e., non-infected) track. However, a cognate mock had been purposely prepared from a separate virus infection experiment alongside its corresponding siRNA blot loading control. We have retrieved all the original data intended to mount Panel 1D and have produced a corrected panel. The correction has no bearing on the original conclusions that distinct single Dicer mutations affect specifically the accumulation of some virus-derived siRNAs, but that these changes in siRNA accumulation cause no noticeable effects on virus (TRV-PDS) RNA accumulation.

Figure 1E depicts TRV-PDS infections of various combination Arabidopsis silencing mutants. The published panel was inappropriately mounted with the wrong ribosomal RNA loading control. In the mounting, this has caused a dcl4 mutant track to be mistakenly used in place of a wild-type track. A vertical line to signal gel splicing did not separate this lane. We have retrieved all the original data intended to mount Panel 1D and have produced a corrected panel. The correction has no bearing on the original conclusions that distinct double Dicer mutations affect the accumulation of specific virus-derived siRNA species, revealing a hierarchical action among the Dicer enzymes. In particular, the dcl2-dcl4 double mutation markedly enhances TRV-PDS accumulation and concomitantly eliminates virus-induced gene silencing of PDS (panel 1L), highlighting the key, redundant contribution of DCL2 and DCL4 to antiviral RNAi through the production of 22-nt and 21-nt virus-derived siRNA, respectively.

Figure 2A is a composite of several separate Northern blots and was found to be erroneously assembled:

(i) The published left panel of Fig. 2A is a section of an original TCV siRNA Northern Blot for which the wrong rRNA loading control was used. We have retrieved the correct loading control for this panel and have amended it accordingly, also specifying the correct separation of samples according to loading control preparation. The amendment does not alter the original conclusion that TCV-derived siRNAs are mostly 22-nt in length, and produced in a strict DCL2-dependent manner.

(ii) The published right panel of Fig. 2A was incorrect as it contained the wrong rRNA loading control and incorporated tracks from the original siRNA blot not corresponding to the labels indicated on the panel. We have retrieved all the original data intended to mount this panel and its cognate loading control and have produced a corrected panel. Despite the nature of the mistake, the amended panel does not contradict the original conclusions drawn from the analysis of single dicer mutants (Fig. 2A, left panel) and confirms that DCL2 is the sole contributor to TCV-derived siRNA production. In particular, the dcl3-dcl4 double mutant, where DCL2 is still active, does accumulates high levels of 22-nt siRNAs, the diagnostic products of DCL2.

(iii) The published bottom panel of Fig. 2A, which depicts a Western analysis of the TCV P38 protein, was initially split to accommodate the wrong loading controls of each upper panel. We have retrieved the original Western blot and loading controls, and have produced a corrected panel. The data from the amended P38 Western blot concur with the original conclusions that none of the single or double dicer mutations affects TCV accumulation noticeably.

Three panels in the Supplementary Materials were also affected. A revised SM file has been posted with the corrected figures, described here.

Figure S1D had a mounting error in which a cognate siRNA panel (upper panel, comparing TRV-derived siRNA species in various rdr mutants) was mistakenly combined with the wrong viral RNA Northern blot and loading controls. We have retrieved all the original data intended for panel D and have produced a corrected panel. The results in the amended figure show that none of the single rdr mutations of Arabidopsis affects the accumulation of TRV-PDS, which is in agreement with the original statement made in reference to Figure S1D. Thus, the mounting error and its correction have no bearing on the original conclusions that single rdr mutations do not overtly affect TRV-PDS accumulation or virus-derived siRNA production.

Figure S1F did not depict the Northern blot intended for publication. We have retrieved all the cognate data prepared originally to mount panel F and have produced a corrected panel. The correction has no bearing on the original conclusion that TRV-PDS accumulates more in the dcl2-dcl4 double mutants, in which virus-induced gene silencing of PDS is concomitantly alleviated. Virus accumulation is even more pronounced in the dcl2-dcl3-dcl4 triple mutant but sporadic VIGS is observed in that case, again agreeing with the original statements.

Figure S2A was assembled with the wrong Northern blot. We have retrieved all the cognate data prepared originally to mount panel A relating to the absence of detectable effects of single rdr mutations on TCV siRNA accumulation. These data were part of the original blots used to correct Fig. 2A (left and right). The correction has no bearing on the original conclusions.

Navigate This Article