Fig. 9
From: A protein complex required for polar growth of rhizobial infection threads
EXO70H4 co-localizes with VPY. a–d Expression of EXO70H4 in roots inoculated by rhizobia (a), at a rhizobial infection site (b), in a root hair containing an infection thread (c) and in nodules (d) as shown by pEXO70H4:GUS activity. e Subcellular localization of GFP-EXO70H4 in root hairs without inoculation with rhizobia. f–i Subcellular localization of GFP-EXO70H4 in root hairs following inoculation with Rm1021-CFP (magenta). j Co-localization of VPY-GFP (green) and mCherry-EXO70H4 (red) at the same punctus in root hairs of rhizobia-inoculated composite plants transformed with pVPY:VPY-GFP / pLjUBQ1:mCherry-EXO70H4. k Bimolecular Fluorescence Complementation (BiFC) experiments in root hairs following inoculation with rhizobia. Split Venus fluorescent protein was used, with N-Venus fused to EXO70H4 driven by pLjUBQ1 promoter and C-Venus fused to VPY, MSP domain or ankyrin repeats containing domain of VPY driven by VPY promoter. l Live cell confocal images showing a root hair undergoing infection in a sunn-2 composite plant containing pVPY:VPY-C-Venus, and pLjUBQ1:N-Venus-EXO70H4. Split Venus fluorescence was imaged 3d after inoculation with Sm2011-CFP, in a root hair hosting an elongating infection thread. Venus fluorescence (arrowheads in l) was associated notably to the tip of a growing infection thread (arrows); Venus fluorescence (open arrowheads) was also found associated to a number of other puncta in the nucleus vicinity. Five ongoing infection sites where imaged in four plants in l. GUS staining is shown as blue. DsRED was used as a transgenic marker. DsRED, Venus, GFP and CFP were given pseudo coloured in red, yellow, green and magenta respectively. n, nucleus. Arrowheads, GFP/mCherry /Venus puncta. Arrows, CFP tagged rhizobia. Scale bars, 100 µm (a, d) and 10 µm (b, c, e–l)
