Fig. 6: Overexpression of VPY in ipd3 ipd3l results in a dmi3-like phenotype and VPY interacts with DMI3. | Nature Communications

Fig. 6: Overexpression of VPY in ipd3 ipd3l results in a dmi3-like phenotype and VPY interacts with DMI3.

From: Distinct ankyrin repeat subdomains control VAPYRIN locations and intracellular accommodation functions during arbuscular mycorrhizal symbiosis

Fig. 6

a–c ipd3-2 ipd3l-2 roots transformed with 35Spro:VPY or a negative control, 35Spro:GFP, colonized with D. epigeae, and harvested three weeks post inoculation. Roots were stained with WGA-AlexaFluor488 (green) and propidium iodide (red) to reveal the fungus and plant cell walls, respectively. Roots overexpressing VPY show hyphopodia that fail to enter the epidermis. Scale bar, 500 μm. c Quantification of surface hyphopodia, infection units, and infection units containing arbuscules in ipd3-2 ipd3l-2 roots overexpressing 35Spro:GFP (n = 6 root systems) and 35Spro:VPY (n = 11 root systems). ** indicates p < 0.01, *** indicates p < 0.001 using a two-tailed Student’s t test, 35Spro:VPY as compared to 35Spro:GFP. Lines in boxplots represent the median value, box limits represent the upper and lower quartiles, whiskers represent 1.5 times the interquartile range. d VPY and DMI3 interact in a co-immunoprecipitation analysis in N. benthamiana. VPY-HA and either GFP-DMI3 or GFP-∆DELLA1 or GFP were co-expressed in N. benthamiana leaves using 35Spro and GFP-tagged proteins were immunoprecipitated using anti-GFP magnetic trap beads. The presence of VPY-HA was assessed via western blot. VPY-HA co-immunoprecipitated with GFP-DMI3 but not with GFP-∆DELLA1 or GFP. e VPY-HA and VPYANK∆3-5-HA were co-expressed with GFP-DMI3 or GFP in N. benthamiana leaves under the 35Spro and GFP-tagged proteins were immunoprecipitated using anti-GFP magnetic trap beads. The presence of VPY-HA and VPYANK∆3-5-HA was assessed by western blot. In both (d) and (e), co-immunoprecipitations were repeated once with similar results.

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