Fig. 2

Detection of GFP in stable transgenic soybeans. a Leaves of T1 lines 131 transformed with the pGmCHIT1:GFP construction were observed using a dissection scope (Leica Z16 APO) under GFP filter and bright light at 24 and 72 h after. P. pachyrhizi inoculation (+) or mock treatment (−). Arrows indicate the inoculation spots. Bar-scales represent 200 μm. b Relative expression of GFP in line 131 (pGmCHIT:GFP) 72 h after P. pachyrhizi inoculation (+) and in non-infected (−) plants. The actin (GenBank: NM_001289231.2) and an unknown protein (GenBank: BE330043) [32] encoding genes were used as references. Three independent biological replicates ± standard errors are shown. *: significant difference between treated (+) and untreated (−) leaves determined by a Student’s t-test (p < 0.05). (c) Detection of GFP protein 72 h after. P. pachyrhizi inoculation in WT and 131 line plants by immunoblotting with an antibody raised against the GFP. Homogenous loading was checked on the gel by Strain Free detection technology (Biorad, US) (below)