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. 2020 Jul 28;10:12630. doi: 10.1038/s41598-020-69417-w

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© The Author(s) 2020

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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(a) Growth of the B. bassiana wild type (WT), Δpks15 and transformants G1, G2 and G6 (from genetic complementation of ∆pks15) on PDA with/without hygromycin B or on a minimal medium containing glufosinate. The three isolates G1, G2 and G6 were resistant to hygromycin B but became sensitive to glufosinate. (b) Virulence against beet armyworm larvae determined by cumulative insect mortalities (%) caused by the B. bassiana wild type, Δpks15 and complemented isolate G6 using a low-dose inoculum (1 × 10⁵ conidia ml⁻¹). Saline was used as the control. (c) Phagocytic assay using the soil amoeba A. castellanii. Mortality rates (%) of the amoebae (A) after incubation with blastospores of the B. bassiana wild type, ∆pks15 and the complemented isolate G6 and S. cerevisiae cells (control). Data shown are mean ± S.E.M. Asterisks indicate statistical significance between the wild type or the complemented isolate G6 and Δpks15 (Student’s t test: *p < 0.05; **< 0.01). (d) Light micrographs of amoebae (A) after incubation with blastospores of the B. bassiana wild type, ∆pks15 and G6 and S. cerevisiae cells for 24 h. The wild type and G6 grew to generate hyphae at this early time point. Bars, 10 µm.