Bidirectional cross-kingdom rnai and fungal uptake of external rnas confer plant protection

ABSTRACT Aggressive fungal pathogens such as _Botrytis_ and _Verticillium_ spp. cause severe crop losses worldwide. We recently discovered that _Botrytis cinerea_ delivers small RNAs

(_Bc_–sRNAs) into plant cells to silence host immunity genes. Such sRNA effectors are mostly produced by _Botrytis cinerea_ Dicer-like protein 1 (_Bc_-DCL1) and _Bc_-DCL2. Here we show that

expressing sRNAs that target _Bc-DCL1_ and _Bc-DCL2_ in _Arabidopsis_ and tomato silences _Bc-DCL_ genes and attenuates fungal pathogenicity and growth, exemplifying bidirectional

cross-kingdom RNAi and sRNA trafficking between plants and fungi. This strategy can be adapted to simultaneously control multiple fungal diseases. We also show that _Botrytis_ can take up

external sRNAs and double-stranded RNAs (dsRNAs). Applying sRNAs or dsRNAs that target _Botrytis DCL1_ and _DCL2_ genes on the surface of fruits, vegetables and flowers significantly

inhibits grey mould disease. Such pathogen gene-targeting RNAs represent a new generation of environmentally friendly fungicides. Access through your institution Buy or subscribe This is a

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DOUBLE-STRANDED RNA PREVENTS AND CURES INFECTION BY RUST FUNGI Article Open access 06 December 2023 CONTROL OF WHITE MOLD (_SCLEROTINIA SCLEROTIORUM_) THROUGH PLANT-MEDIATED RNA INTERFERENCE

Article Open access 20 April 2023 HIGH-EFFICIENCY GREEN MANAGEMENT OF POTATO LATE BLIGHT BY A SELF-ASSEMBLED MULTICOMPONENT NANO-BIOPROTECTANT Article Open access 12 September 2023

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expression analysis. _Nat. Protoc._ 2, 1565–1572 (2007). Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We thank H. Vaucheret for the _ago1-27_ seeds, M. Coffey for the

_V. dahliae JR2_ strain, I. Kaloshian for providing the growth room space for VIGS experiments, and Y. Lii for editing the paper. This work was supported by grants from National Institute of

Health (R01 GM093008), National Science Foundation (IOS-1257576, IOS-1557812) and an AES-CE Award (PPA-7517H) awarded to H.J. AUTHOR INFORMATION Author notes * Arne Weiberg Present address:

†Present address: Institute of Genetics, University of Munich, Martinsried, Germany., AUTHORS AND AFFILIATIONS * Department of Plant Pathology and Microbiology, Center for Plant Cell

Biology, Institute for Integrative Genome Biology, University of California, Riverside, 92521, California, USA Ming Wang, Arne Weiberg & Hailing Jin * Institute of Bioinformatics and

Systems Biology, National Chiao Tung University, 300, Hsin-Chu, Taiwan Feng-Mao Lin & Hsien-Da Huang * Laboratory of Phytopathology, Wageningen University, Droevendaalsesteeg 1, 6708 PB

Wageningen, the Netherlands Bart P. H. J. Thomma Authors * Ming Wang View author publications You can also search for this author inPubMed Google Scholar * Arne Weiberg View author

publications You can also search for this author inPubMed Google Scholar * Feng-Mao Lin View author publications You can also search for this author inPubMed Google Scholar * Bart P. H. J.

Thomma View author publications You can also search for this author inPubMed Google Scholar * Hsien-Da Huang View author publications You can also search for this author inPubMed Google

Scholar * Hailing Jin View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS H.J. conceived the idea. M.W. and H.J. designed the experiments. M.W.

performed most of the experiments and analysed data. A.W. profiled the sRNAs from _dcl1 dcl2_ and WT strains and analysed the data. F.M.L. and H.D.H. conducted bioinformatics analysis on

sRNA libraries. B.T. provided Vd genome sequence for JR2 strain. M.W., A.W. and H.J. wrote the manuscript. CORRESPONDING AUTHOR Correspondence to Hailing Jin. ETHICS DECLARATIONS COMPETING

INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary Methods, Supplementary Figures 1–9, Supplementary References

(PDF 3132 kb) SUPPLEMENTARY TABLE 1 The normalized read counts of previously predicted _Bc-s_RNA effector candidates in _B. cinerea WT_ and _dcl1 dcl2_ strains (XLSX 76 kb) SUPPLEMENTARY

TABLE 2 At-AGO1-associated _Vd_-sRNA effector candidates and their targets. (XLSX 36 kb) SUPPLEMENTARY TABLE 3 At-AGO2-associated _Vd_-sRNAs and their targets. (XLSX 13 kb) SUPPLEMENTARY

TABLE 4 The list of primers and oligoes used in the manuscript. (XLSX 51 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Wang, M., Weiberg, A., Lin,

FM. _et al._ Bidirectional cross-kingdom RNAi and fungal uptake of external RNAs confer plant protection. _Nature Plants_ 2, 16151 (2016). https://doi.org/10.1038/nplants.2016.151 Download

citation * Received: 26 December 2015 * Accepted: 01 September 2016 * Published: 19 September 2016 * DOI: https://doi.org/10.1038/nplants.2016.151 SHARE THIS ARTICLE Anyone you share the

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