Fig. 3: The expression pattern of GmRR11d during nodulation.

a, b qRT-PCR analysis of GmRR11d expression in different tissues or during nodulation stages. a Roots, leaves, and nodules of soybean seedlings inoculated with B. diaefficiens strain USDA110 were harvested at 28 DAI and used for gene expression analyses. GmELF1b was used as the endogenous control gene. Data are presented as means ± SD from three biological replicates. More than 6 samples were analyzed in each independent biological repeat. Different letters indicate significant differences at p < 0.05 (One-way ANOVA). b Seven-day-old seedlings were inoculated with B. diaefficiens strain USDA110 and the infected root materials were collected at 0, 1, 3, 6, 12 HAI and 1, 3, 6,10, 14, 21, 28 DAI for expression analysis of GmRR11d. GmELF1b was used as the endogenous control gene. The data are shown as the means ± SD from three biological replicates. Asterisks indicate significant differences relative to 0 HAI (Two-sided Student’s t-test, *p < 0.05; **p < 0.01; ***p < 0.001). c–p Histochemical assay of proGmRR11d:GUS and in the transgenic roots and nodules. Three independent experiments were repeated with similar results. c–j Representative GUS staining in hairy roots of proGmRR11d:GUS. GUS activity was monitored at 12 HAI c, d, 1 DAI e, f, 3 DAI g, h and 6 DAI i, j with B. diazoefficiens USDA110. Scale bars = 100 μm in c–j. k–n The expression pattern of GmRR11d in emerging nodule k, young nodule l and mature nodule m, n. o, p Cross-sectional analysis of the expression pattern of GmRR11d in young and mature nodules, the red arrows show cortex. Scale bars = 1 mm in k–p.