Figure 3 | Scientific Reports

Figure 3

From: Rhizobia use a pathogenic-like effector to hijack leguminous nodulation signalling

Figure 3

Expression, secretion, translocation and localization of the B. elkanii type III effector Bel2-5. (a) RT-qPCR analysis of bel2-5 and nopA expressions in wild-type and ttsI mutant (BEttsI) of B. elkanii USDA61. Total RNAs were isolated from cultures in the absence (−) or presence (+) of the genistein (10 µM) after 48 h induction. The expression level of each gene was normalized by the ATP synthase (atpD) gene. Values represent mean ± SD (n = 3). (b) Secretion of Bel2-5 and NopA in the culture supernatant of B. elkanii strains. Secreted proteins from culture supernatants were subjected to immunoblot analysis with the anti-Bel2-5 (α-Bel2-5) or anti-NopA (α-NopA) antibodies. Molecular masses (kDa) of the marker are shown on the left. (c) Translocation of Bel2-5 into soybean nodules. cAMP levels measured from G. max cv. Enrei nodules harvested at 18 dpi. Plants were inoculated with B. elkanii USDA61 wild-type, BECya and BErhcJCya. The USDA61 wild-type contained no cya fusion. Values represent mean ± SD from duplicates measurements. (d) Bel2-5 is targeted to the plant nuclei. Bel2-5 protein fused with eGFP was transiently expressed in N. benthamina leaves by Agrobacterium infiltration and visualized 48 h after by confocal microscopy to determine the subcellular localization. From left to right: an overlay of GFP and DAPI fluorescence spectrums, and the GFP and DAPI fluorescence spectrum, respectively. Staining with DAPI was used to visualize nuclei. On the overlay panel, the DAPI fluorescence was displayed in gray and the contrast and brightness were enhanced in order to be able to visualize the contour of the cells. Scale bars: 100 µm. Means followed by different letters are significantly different at the 0.05 level by Tukey’s method (a,c).

Back to article page