A genetic screen for impaired systemic RNAi highlights the crucial role of DICER-LIKE 2

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dc.contributor.authorTaochy, C.
dc.contributor.authorGursanscky, N.
dc.contributor.authorCao, J.
dc.contributor.authorFletcher, S.
dc.contributor.authorDressel, U.
dc.contributor.authorMitter, N.
dc.contributor.authorTucker, M.
dc.contributor.authorKoltunow, A.
dc.contributor.authorBowman, J.
dc.contributor.authorVaucheret, H.
dc.contributor.authorCarroll, B.
dc.date.issued2017
dc.description.abstractPost-transcriptional gene silencing (PTGS) of transgenes involves abundant 21 nt small interfering RNAs (siRNAs) and low abundance 22 nt siRNAs produced from double-stranded RNA (dsRNA) by DCL4 and DCL2, respectively. However, DCL2 facilitates the recruitment of RNA-DEPENDENT RNA POLYMERASE 6 (RDR6) to ARGONAUTE 1 (AGO1)-derived cleavage products, resulting in more efficient amplification of secondary and transitive dsRNA and siRNAs. Here, we describe a reporter system where RDR6-dependent PTGS is initiated by restricted expression of an inverted-repeat dsRNA specifically in the root tip, allowing a genetic screen to identify mutants impaired in RDR6-dependent systemic PTGS. Our screen identified dcl2 but not dcl4 mutants. Moreover, grafting experiments showed that DCL2, but not DCL4, is required in both the source rootstock and recipient shoot tissue for efficient RDR6-dependent systemic PTGS. Furthermore, dcl4 rootstocks produced more DCL2-dependent 22 nt siRNAs than wild type and showed enhanced systemic movement of PTGS to grafted shoots. Thus, along with its role in recruiting RDR6 for further amplification of PTGS, DCL2 is crucial for RDR6-dependent systemic PTGS.
dc.description.statementofresponsibilityChristelle Taochy, Nial R. Gursanscky, Jiangling Cao, Stephen J. Fletcher, Uwe Dressel, Neena Mitter, Matthew R. Tucker, Anna M.G. Koltunow, John L. Bowman, Hervé Vaucheret and Bernard J. Carroll
dc.identifier.citationPlant Physiology, 2017; 175(3):1424-1437
dc.identifier.doi10.1104/pp.17.01181
dc.identifier.issn0032-0889
dc.identifier.issn1532-2548
dc.identifier.orcidTucker, M. [0000-0003-4661-6700]
dc.identifier.urihttp://hdl.handle.net/2440/110569
dc.language.isoen
dc.publisherAmerican Society of Plant Biologists
dc.relation.granthttp://purl.org/au-research/grants/arc/DP0988294
dc.relation.granthttp://purl.org/au-research/grants/arc/DP120103966
dc.relation.granthttp://purl.org/au-research/grants/arc/DP150104048
dc.relation.granthttp://purl.org/au-research/grants/arc/DP160100892
dc.rights© 2017 American Society of Plant Biologists. All Rights Reserved.
dc.source.urihttps://doi.org/10.1104/pp.17.01181
dc.subjectArabidopsis
dc.subjectPlant Shoots
dc.subjectPlant Roots
dc.subjectRibonuclease III
dc.subjectGlucuronidase
dc.subjectCell Cycle Proteins
dc.subjectGreen Fluorescent Proteins
dc.subjectArabidopsis Proteins
dc.subjectRNA, Small Interfering
dc.subjectRNA Interference
dc.subjectPhenotype
dc.subjectMutation
dc.subjectGenes, Reporter
dc.subjectModels, Biological
dc.subjectGenetic Testing
dc.subjectRNA-Dependent RNA Polymerase
dc.titleA genetic screen for impaired systemic RNAi highlights the crucial role of DICER-LIKE 2
dc.typeJournal article
pubs.publication-statusPublished

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