| Autor: |
Claar Martina, Weipert Lisa, Garikapati Vannuruswamy, Busche Tobias, Biedenkopf Dagmar, Barth Patrick, Hardt Martin, Möbus Anna, Werner Bernhard Timo, Kalinowski Jorn, Schlemmer Timo, Koch Aline, Spengler Bernhard, Jelonek Lukas, Preußer Christian, Goesmann Alexander |
| Rok vydání: |
2020 |
| Předmět: |
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| DOI: |
10.1101/2020.02.12.945154 |
| Popis: |
Small (s)RNAs and their double-stranded (ds)RNA precursors have been adopted to control diseases in crop plants through expression in transgenic plants and targeted gene silencing (host-induced gene silencing, HIGS). While HIGS strategies proved to be effective, the mechanism of RNA transfer at the plant - pathogen interface is widely unknown. Here we show that extracellular vesicles (EVs) purified fromArabidopsis thalianaplants expressing CYP3RNA, a dsRNA originally designed to target the threeCYP51genes of the fungal pathogenFusarium graminearum, contain CYP3RNA-derived small interfering (si)RNAs as shown by RNA sequencing (RNA-seq) analysis. These transgene specific siRNAs had a length of 21 and 22 nucleotides with a bias towards 5’-uracil (U) and 5’-adenine (A). Notably, stringent protease and RNase treated EV fractions contained >70% less CYP3RNA-derived siRNAs, suggesting the presence of co-purified extravesicular nucleoprotein complexes stabilizing siRNAs outside of EVs. In addition, mutants of the ESCRT-III complex showed a loss of HIGS-mediated disease resistance and EVs isolated from these mutants were free of CYP3RNA-derived siRNAs. Together, these findings support the view that endosomal vesicle trafficking is required for HIGS mediating the transfer of transgene-derived siRNAs between donor host cells and recipient fungal cells probably in an EV-independent manner. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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