Extended Data Fig. 9: PACEs from FaFaCuRo species are functionally equivalent in restoring bacterial infection in the L. japonicus nin-15 mutant.
From: A novel cis-element enabled bacterial uptake by plant cells
L. japonicus nin-15 roots were transformed with T-DNAs carrying a Ubq10pro:NLS-GFP transformation marker in tandem with the LjNIN gene driven by either of the following promoters: (A) the 3 kb LjNIN promoter (NINpro), the LjNIN minimal promoter (NINminpro), the 3 kb LjNIN promoter with PACE deleted (NINpro::∆PACE) or mutated (NINpro::mPACE); (B) the 3 kb LjNIN promoter with LjPACE replaced with either of the PACE sequence variants from nodulating or non-nodulating FaFaCuRo species and analysed 21 dpi with M. loti DsRed. (A – B) Representative overview pictures of nin-15 transgenic roots systems. Sections of representative nodules are displayed in Fig. 3. Note the drastic reduction of restoration of infection in nodules and root hairs associated with the mutation or deletion of PACE as well as the replacement of PACE with JrPACE-like in the context of the LjNIN promoter. White asterisks and arrowheads: infected and non-infected nodules, respectively. (C – E) Boxplots displaying (C) the percentage of root hair ITs among total infection events (sum of bacterial entrapments and ITs) and (D – E) the number of infected nodules from two independent experiments. Each dot represents one nin-15 transgenic root piece (C) or root system (D – E). (C) displays merged data from experiments in (D – E) as the percentage represents a normalised value calculated for each root piece (see Supplementary Table 4). Thick black lines, median; box, interquartile range; whiskers, lowest and highest data point within 1.5 interquartile range (IQR); black filled circles, data points inside 1.5 IQR; white filled circles, data points outside 1.5 IQR of the upper/lower quartile. n: number of transgenic root systems or root pieces analysed. For species abbreviations see Extended Data Fig. 2a. The applied statistical method was ANOVA with post hoc Tukey: (C) F9,313 = 106.7, p < 2×10−16; (D) F6,346 = 82.89, p < 2×10−16; (E) F4,135 = 20.18, p = 4.76×10−13. Different small letters indicate significant differences. Data are from a single experiment. Bars, 2 mm. WLI: white light illumination.
