Fig. 4: Alterations of GmRR11d expression affect nodulation.

a–c Overexpression of GmRR11d reduces nodule number. a qRT-PCR analysis of transgenic hairy roots expressing 35 S:GmRR11d. The expression levels were normalized against the geometric mean of soybean GmELF1b. Data are presented as means ± SD from three biological replicates. Asterisks indicate significant differences relative to the EV1 control. Two-sided Student’s t-test ***p 0.001. b, c Quantification and representative images of nodule number per hairy root expressing EV1 and 35 S:GmRR11d. Data are presented as means ± SD (14 hairy roots in EV1 and 18 hairy roots in 35 S:GmRR11d). Three independent experiments were repeated with similar results. Asterisks indicate significant differences relative to the EV1 control. Two-sided Student’s t-test, ***p < 0.001. Scale bars = 1 cm. d–f Knockdown of GmRR11d increases nodule number. d qRT-PCR analysis of transgenic hairy roots expressing RNAi-GmRR11d. The expression levels were normalized against the geometric mean of soybean GmELF1b. Data are presented as means ± SD from three biological replicates. Asterisks indicate significant differences relative to the EV2 control. Two-sided Student’s t-test, *p < 0.05. e, f Nodulation phenotypic analysis of individual hairy roots expressing EV2 and RNAi-GmRR11d at 21 DAI. Scale bars = 1 cm. Data are presented as means ± SD (24 hairy roots in EV2 and 28 hairy roots in RNAi-GmRR11d). Three independent experiments were repeated with similar results. Asterisks indicate significant differences relative to the EV2 control. Two-sided Student’s t-test, ***p < 0.001. Boxes indicate the first and third quartiles and the whiskers indicate the minimum and maximum values, the black lines within the boxes indicate the median values and black circles mark the individual measurements. The numbers above the boxes in (b) and (e) represent the mean values.