Extended Data Fig. 9: The RuvC sequence is conserved in several pathogenic bacteria and expression of the XooRuvCPR1 in rice.
a, Sequence alignment of RuvCs in different bacteria. The aspartic acid active site was marked by carmine, and the glutamic acid site was highlighted by dark blue. Pa, Pseudomonas aeruginosa; Pst, Pseudomonas syringae pv. tomato str. DC3000; Ec, Escherichia coli; Rs, Ralstonia solanacearum; Xoo, Xanthomonas oryzae pv. Oryzicola; Bs, Bacillus subtilis. b, Purified His-XooRuvC and His-XooRuvX fusion proteins from E. coli. Expression was induced by IPTG and purified by affinity chromatography. c, XooRuvX treatment did not affect Xoo biofilm. Biofilms were stained with crystal purple and imaged in the PCR tubes. Data are presented as mean values ± SD (n=6 biological replicates). Statistically significant differences in c are indicated by different lowercase letters (One-way ANOVA, P < 0.05). d, Verification of the XooRuvC expression in rice transgenic lines. Total RNA was extracted from the transgenic rice leaves. The XooRuvC gene expression level detected by RT-PCR and ACTIN1 used as an internal control. Band intensity was quantified by Image J. Wild-type Nipponbare was used as a control. e, Gross morphologies of OxXooRuvCPR1 lines and WT in the experimental greenhouse. Scale bars indicate 30 cm. f, Images of seeds harvested from the experimental field. Scale bars indicate 10 mm. g, Hundred-grain weight of seeds. Data are presented as mean values ± SD (n=10 individual plants). The wild type plant Nipponbare was used as the control group. Statistically significant differences are indicated by different lowercase letters (One-way ANOVA, P < 0.05). The above experiments were repeated three times with similar results.
