Fig. 7: GmBTB/POZ weakens GmLHP1-mediated suppression of GmWRKY40 and increases GmLHP1-regulated responses to P. sojae. | Communications Biology

Fig. 7: GmBTB/POZ weakens GmLHP1-mediated suppression of GmWRKY40 and increases GmLHP1-regulated responses to P. sojae.

From: GmBTB/POZ promotes the ubiquitination and degradation of LHP1 to regulate the response of soybean to Phytophthora sojae

Fig. 7

a GmWRKY40 transcript levels in GmBTB/POZ-OE and GmBTB/POZ-RNAi soybean plants. b GmLHP1 transcript levels in EV, GmLHP1-OE, and GmBTB/POZ-OE/GmLHP1-OE transgenic soybean hairy roots. c GmBTB/POZ transcript levels in EV, GmLHP1-OE, and GmBTB/POZ-OE/GmLHP1-OE transgenic hairy roots. d GmWRKY40 transcript levels in EV, GmLHP1-OE, and GmBTB/POZ-OE/GmLHP1-OE transgenic hairy roots. e Relative biomass of P. sojae in EV, GmLHP1-OE, and GmBTB/POZ-OE/GmLHP1-OE transgenic hairy roots based on the transcript level of P. sojae TEF1 (EU079791) after 48 h of P. sojae inoculation. f Subcellular localization of various GmBTB/POZ deletion mutants. Left column, scheme of GmBTB/POZ and its deletion mutants fused to GFP. FL, full length. Right column, the indicated constructs were transiently expressed in Arabidopsis protoplasts and inspected with a confocal microscope. Scale bars indicate 10 µm. g In vitro cell-free degradation assays of GmLHP1-His in protein extracts from GmBTB/POZ-1-OE soybean hairy roots. h GmBTB/POZ-1 promotes the ubiquitination of GmLHP1 in vivo. GmLHP1-Flag was immunoprecipitated using anti-Flag-Tag Mouse mAb (Agarose Conjugated) from GmLHP1-OE and GmLHP1-OE/GmBTB/POZ-1-OE soybean hairy roots by high-efficiency A. rhizogenes-mediated transformation. The transgenic hairy roots were treated with 100 µM MG132 for 8 h before extraction. The immunoprecipitated protein was examined using anti-Flag and anti-ubi antibodies. i GmLHP1 transcript levels in EV, GmLHP1-OE, and GmLHP1-OE/GmBTB/POZ-1-OE transgenic soybean hairy roots. j GmBTB/POZ transcript levels in EV, GmLHP1-OE, and GmLHP1-OE/GmBTB/POZ-1-OE transgenic hairy roots. k GmWRKY40 transcript levels in EV, GmLHP1-OE, and GmLHP1-OE/GmBTB/POZ-1-OE transgenic hairy roots. The housekeeping gene GmEF1 was used as an internal control to normalize the data. The experiment was performed on three biological replicates, each with three technical replicates, and the results were statistically analyzed using Student’s t-test (*P < 0.05, **P < 0.01). Bars indicate the standard deviation of the mean (n = 3).

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