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. 2020 Oct 26;6(4):248. doi: 10.3390/jof6040248

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Pex6 deficiency impacts toxin production in the tangerine pathotype of A. alternata. (A) A leaf necrosis assay for the toxicity of ACT toxin by soaking the petiole of the leaf in cell-free culture filtrates prepared from wild type (WT), two pex6 mutants (Δpex6-M4 and M9) and the complementation strain (CP4) grown in potato dextrose broth for 7 days; (B) Quantitative measurement of necrotic lesions caused by cell-free culture filtrates (panel A) using Image J software. Significance of differences was analyzed using Tukey’s HSD post-hoc test (p ≤ 0.05). Means followed by the same letters are not significantly different; (C) Thin-layer chromatography (TLC) analysis of ACT; (D) High performance liquid chromatography (HPLC) analysis of ACT; (E) A leaf necrosis assay for the toxicity of ACT toxin eluted from HPLC (peaks with retention times of 7.4 and 7.9 min, indicated by a red arrow in panel D) by placing 5 µL elutes at the base of the petiole. The leaves were maintained in a moisture chamber for 3 days for lesion development. Each sample was tested on at least five different leaves and experiments were repeated at least three times.