Fig. 2: Functional validation of cruciferous GSL5s in Arabidopsis.

a, Diagram representation of GSL5 genes from A. thaliana, B. napus, B. rapa, B. oleracea and R. sativus. The allotetraploid B. napus contains two GSL5s in its genome. nt, nucleotide. b, Diagram representation of GSL5 proteins from the above five cruciferous species. aa, amino acid; TMs, transmembrane domains; FKS1, FKS1-like 1,3-beta-glucan synthase domain. c, Clubroot resistance test of the Arabidopsis gsl5 (pmr4-1) mutant. Wild-type plants were used as the susceptible control. Representative photos were taken at 24 dpi with P. brassicae. d, Disease index values of wild-type and gsl5 mutant plants. e, Genetic validation of GSL5 for clubroot susceptibility in Arabidopsis. gsl5-KO, the gsl5 mutant generated by genome editing; GSL5-OE, GSL5-overexpressing plants; gsl5 GSL5, the complemented plants generated by expressing the GSL5 genomic region in the gsl5 mutant. Representative root photos were taken at 24 dpi. f, Disease index values of wild-type, gsl5, gsl5-KO, GSL5-OE and gsl5 GSL5 mutant plants. g, Genetic validation of GSL5s from four cruciferous crops for clubroot susceptibility in Arabidopsis. Representative root photos were taken at 24 dpi. Hap_1, BnaA09.GSL5 Hap_1; Hap_2, BnaA09.GSL5 Hap_2; BnaC09.GSL5, the GSL5 from the C subgenome of B. napus; BraGSL5, the GSL5 from B. rapa; BolGSL5, the GSL5 from B. oleracea; RsaGSL5, the GSL5 from R. sativus L. h, Disease index values of materials in g. i, Resistance test of Arabidopsis gsl5 plants against 36 field P. brassicae isolates (representing 26 pathotypes). Wild-type plants were used as the susceptible control (**P < 0.01). Data in bar graphs are presented as means; error bars, s.d. In d, f and h, statistical significance was determined by a two-sided Student’s t-test with 3 biological replicates in each of 30 plants. Scale bars in c, e and g, 1 cm. In i, statistical significance was determined by the Fisher–Freeman–Halton test; sample sizes and P values are listed in Supplementary Table 7.