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. 2023 Jul 20;14:4383. doi: 10.1038/s41467-023-40093-4

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a B. cinerea sRNAs were examined in B. cinerea EVs isolated by sucrose gradient fractionation. b B. cinerea sRNAs can be protected from micrococcal nuclease treatment by EVs. F4, B. cinerea EV samples were collected from fraction 4 after sucrose gradient centrifugation. Bc-siR17 and Bc-siR9 were used as negative controls. c The Δbcpls1 strain showed strongly reduced virulence, whereas the Δbctsp3 strain showed similar virulence with the wild-type B05 strain. Both Δbcpls1 and Δbctsp3 complementary strains showed similar virulence with the wild-type B05 strain. d Bc-sRNAs were examined by Real-time PCR in EVs isolated from B05, Δbcpls1, Δbctsp3, Δbcpls1 complementary, and Δbctsp3 complementary strains. The data are presented as mean ± s.d. Similar results were obtained in three biologically independent experiments. e Wild-type B. cinerea EVs partially complement the virulence of B. cinerea Δbcpls1 mutant. Premixing of wild-type B. cinerea EVs from sucrose gradient fraction 4 with Δbcpls1 mutant spores partially complement the weak virulence of Δbcpls1 on Arabidopsis leaves. f Wild-type B. cinerea EVs partially complement the virulence of B. cinerea dcl1dcl2 mutant. Premixing of wild-type B. cinerea EVs from sucrose gradient fraction 4 with dcl1dcl2 mutant spores partially complements the weak virulence of dcl1dcl2 on Arabidopsis leaves. Relative lesion size was determined 2 days after infection. The statistical data in c, e and f are presented as mean ± s.d., n = 10 biological replicates. The statistical analysis was performed using ANOVA Dunnett’s multiple comparisons test. The small open circles represent individual values. The error bars indicate s.d. **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.