Identification of plant cadmium resistance gene family in Brassica napus and functional analysis of BnPCR10.1 involved in cadmium and copper tolerance

Abstract

The plant cadmium resistance (PCR) family proteins play important roles in maintaining metal homeostasis and detoxification. However, few functional PCR genes have been well-characterized in plants. In this study, we identified and cloned 26 BnPCR genes from the rapeseed (Brassica napus) genome. They were divided into four groups (I-IV) based on their phylogenetic relationship. Yeast functional complementation experiments showed that BnPCRs can transport copper (Cu) and cadmium (Cd) in yeast. The expression levels of the BnPCRs were variable among different organs. Moreover, most of the genes were induced by Cu²⁺ and Cd²⁺ stress. Among these genes, BnPCR10.1 was highly expressed in various organs and induced by Cu²⁺ and Cd²⁺. Therefore, we studied the function of BnPCR10.1 in more detail. BnPCR10.1 was localized to the plasma membrane (PM), and expression in yeast enhanced yeast cells to export Cu and Cd. Furthermore, overexpression of BnPCR10.1 transgenic lines pro35S::BnPCR10.1;athma5 had lower concentration of Cu in roots than athma5 mutants. In addition, transgenic plants pro35S::BnPCR10.1;atpdr8 had lower concentration of Cd in shoots and roots than atpdr8 mutants. Net Cu²⁺ and Cd²⁺ efflux assay showed that there was decreased absorption of Cu²⁺ and Cd²⁺ in the transgenic Arabidopsis elongation zone of roots than in athma5 and atpdr8 mutants, respectively. These results provide new information on BnPCRs and their roles in response to heavy metals and reveal the mechanism used by BnPCR10.1 to detoxify Cu and Cd. Our findings facilitate a theoretical basis for the genetic improvement of Cu-Cd tolerance in rapeseed.

Keywords: BnPCR10.1; Brassica napus; Cu and Cd tolerance; PCR family; Plasma membrane.