Loss of PMR4 callose synthase results in salicylic acid-independent and broad-spectrum resistance to clubroot in Arabidopsis and Brassica napus

The clubroot disease caused by protist Plasmodiophora brassicae is one of the most important diseases of Brassica crops. Growing clubroot-resistant cultivars remains the most effective and practical approach to managing clubroot disease. However, resistance gene-mediated immunity is rapidly overcome in the field when new pathotypes arise. In this study, we identified PMR4 as a potential gene target for creating a novel clubroot-resistant source. Recessive PMR4 mutations in Arabidopsis thaliana conferred broad-spectrum resistance to multiple P. brassicae pathotypes, independent of salicylic acid-mediated plant immunity. CRISPR/Cas9-mediated gene-editing was employed to create mutations in two PMR4 orthologs in the B. napus genome, and resulting homozygous mutants exhibited dual resistance to powdery mildew and clubroot. PMR4 is required for the callose deposition at wound and powdery mildew infection sites in leaves. This study reveals that callose deposition in roots is induced by P. brassicae infection and requires PMR4. It appears that the clubroot disease progression is arrested at the primary-to-secondary infection phase in pmr4-1 mutants. Together, this study demonstrates that PMR4 -encoded callose synthase is a host susceptibility factor required for P. brassicae to complete its life cycle, and that PMR4 can be targeted against both powdery mildew and clubroot diseases in Brassica crops.