Correction: Kim et al. The Auto-Regulation of ATL2 E3 Ubiquitin Ligase Plays an Important Role in the Immune Response against Alternaria brassicicola in Arabidopsis thaliana. Int. J. Mol. Sci. 2024, 25, 2388

Abstract

In the original publication [...].

Figure 4

Requirement of ATL2 E3 ligase activity for disease response to A. brassicicola. (A) The activity of ATL2 E3 is dependent on the presence of the Cys138 residue in the RING domain. To analyze self-ubiquitination assays of MBP-ATL2 or MBP-ATL2^C138A proteins in the presence of rabbit E1, human E2, and Ub, Western blot analyses were conducted using anti-MBP and anti-Ub antibodies. Arrows denote the non-ubiquitinated forms of the proteins. (B) RT-PCR analysis of ATL2 transcripts of Col-0, ATL2^C138AOX2-1-, and ATL2^C138AOX7-5-overexpressing plants. ACTIN2 was used as a loading control. (C) The presence of the C138A mutation in the RING domain of ATL2 is necessary for a disease response to A. brassicicola. Photographs of the leaves of Col-0, ATL2OX4-1-, ATL2^C138AOX2-1-, and ATL2^C138AOX7-5-overexpressing plants post-infection with A. brassicicola are shown. Detached leaves from 6-week-old plants were inoculated with a 5 µL droplet of A. brassicicola spores (approximately 5 × 10⁵ spores/mL), and photographs were taken 6 days after inoculation. The white arrows indicate inoculated regions. (D) Size of lesions in A. brassicicola infected Col-0, ATL2OX4-1-, ATL2^C138AOX2-1-, and ATL2^C138AOX7-5-overexpressing plant leaves. Lesion size was measured at 6 days after inoculation. (E) Trypan blue staining showing fungal hyphae of A. brassicicola on Arabidopsis leaves; Col-0, ATL2OX4-1, ATL2^C138AOX2-1, and ATL2^C138AOX7-5. Scale bars = 50 µm. (F) Measurement of spore numbers in lesions. The spore numbers in lesions were measured by collecting total spores from detached inoculated leaves and counting them microscopically at 6 days after inoculation. The data are shown as mean ± SD (n = 3), with ** p < 0.005, and *** p < 0.001 indicating the significance level. In panels D and F, the p-value indicates the significance relative to the A. brassicicola treatment at wild-type (Col-0) plants and was determined and analyzed using GraphPad Prism 8 via two-way ANOVA followed by Dunnett’s multiple comparison. n.s represents not significant. These experiments were repeated three times (biological replicates) with similar results.