Fig. 1. AtALA1 is responsible for the vacuolar compartmentation of DON, and AtALA7 is responsible for that of CIA.

a DON tolerance assay of wild-type, and AtALA1 mutant (ala1, ala1-8) and complemented (Comp1, Comp1-8) Arabidopsis seedlings. The native promoter-controlled AtALA1 was used to rescue the defect of AtALA1 in mutants. Six-day-old seedlings were transferred to MS media containing DON (50 μg/mL) for 7 d. WT, wild-type; ala1 and ala1-8, AtALA1 loss-of-function mutant. Comp1 and Comp1-8, complemented ala1 and ala1-8 seedlings expressing proAtALA1::EGFP-AtALA1. DON, deoxynivalenol; proAtALA1, the native promoter of the AtALA1 gene. Scale bar, 1 cm. b Proportion of bleached leaves suffering from DON. Data are presented as box-and-whisker plots of three replicates (eight plants each). c Weight of plants with or without DON treatment. Data are presented as box-and-whisker plots of three replicates (six plants each). d Distribution of DON^5-FAM in root cells of wild-type, ala1 mutant, and complemented Arabidopsis seedlings. Seedlings were treated with DON^5-FAM (9 μg/mL) and FM4-64 (8 μM) for 2 h, 6 h, and 10 h. Confocal images at 2 h were collected from root hair cells, whereas those at 6 h and 10 h were collected from cells of the root meristem zone. At least three independent repeats were performed. DON^5-FAM, DON labeled with 5-carboxyfluorescein (5-FAM). Scale bar, 10 μm. e Percentage of cells containing DON^5-FAM in their vacuoles at 10 h. Data are presented as dot plots (n = 12 roots). f CIA tolerance assay of wild-type, AtALA7 mutant (ala7, ala7-24) and complemented (Comp7, Comp7-24) Arabidopsis seedlings. The native promoter-controlled AtALA7 was used to rescue the defect of AtALA7 in mutants. Six-day-old seedlings were transferred to MS media containing CIA (100 μg/mL) for 7 d. ala7 and ala7-24, AtALA7 loss-of-function mutant. Comp7 and Comp7-24, complemented ala7 and ala7-24 mutants seedlings expressing proAtALA7::AtALA7-eGFP. CIA, cinnamate acetate; proAtALA7, the native promoter of the AtALA7 gene. Scale bar, 1 cm. g Proportion of bleached leaves resulting from CIA treatment. Data are shown as box-and-whisker plots of three replicates (eight plants each). h Weight of plants with or without CIA treatment. Data are presented as box-and-whisker plots of three replicates (six plants each). i Distribution of CIA^FITC in root cells of wild-type, ala7 mutant and complemented (Comp7) Arabidopsis seedlings. The seedlings were treated with CIA^FITC (5 μg/mL) and FM4-64 (8 μM) for 1 h, 2 h, and 8 h. Confocal images at 1 h and 2 h were collected from the root hair cells, while those at 10 h were collected from cells of the root meristem zone. At least three independent repeats were conducted. CIA^FITC, CIA labeled with fluorescein-isothiocyanate-isomer (FITC). Scale bar, 0.5 μm. j Percentage of cells containing CIA^FITC in their vacuoles at 8 h. Data are presented as dot plots (n = 12 roots). Box-and-whisker plots show the medians (horizontal lines), upper and lower quartiles (box edges), and 1.5× the interquartile range (whiskers). Different letters in b, c, e, g, h, and j represent significant differences at P < 0.05 by one-way ANOVA with Tukey multiple comparisons test. n.s., not significant.