The root meristem growth factor BrRGF6 positively regulates Chinese cabbage to infection of clubroot disease caused by Plasmodiophora Brassicae

Abstract

Chinese cabbage has a high annual demand in China. However, clubroot disease caused by the infection of Plasmodiophora brassicae seriously affects its yield. Transcriptome analysis identified a root meristem growth factor 6 (BrRGF6) as significantly up-regulated in Chinese cabbage roots infected with Plasmodiophora brassicae. Quantitative reverse-transcription polymerase chain reaction and in situ hybridization analysis showed higher BrRGF6 expression in susceptible materials than in resistant materials. After Plasmodiophora brassicae infection, BrRGF6 expression was significantly up-regulated, especially in susceptible materials. Gene function analysis showed that the roots of Arabidopsis mutant rgf6 grew faster than the wild-type, and delayed the infection progress of Plasmodiophora brassicae. A Protein, nuclear transcription factor Y subunit C (BrNF-YC), was screened from yeast two-hybrid library of Chinese cabbage induced by Plasmodiophora brassicae, and verified to interact with BrRGF6 by yeast two-hybrid co-transfer. Yeast one-hybrid and β-Glucuronidase activity analysis showed that BrNF-YC could directly bind to and strongly activate the promoter of BrRGF6. Transgenic verification showed that BrRGF6 or BrNF-YC silenced Chinese cabbage significantly decreased the expression of BrRGF6, accelerated root development, and reduced incidence of clubroot disease. However, after overexpression of BrRGF6 or BrNF-YC, the phenotype showed a reverse trend. Therefore, BrRGF6 silencing accelerated root growth and enhanced resistance to clubroot disease, which was regulated by BrNF-YC.