Enhanced expression of the rice gene OsCTF using epigenetic regulation for low cadmium planting in Rice

Abstract

Rice is a major food crop and one of the most cadmium absorbing and bulk cereal crops. OsCTF, a gene involved in the efflux of heavy metal Cd in rice, which is closely related to the amount of Cd accumulation in rice. In this study, CRISPR-dCas9 technology was used to regulation of OsCTF 3 ‘- UTR demethylation in rice to improve the amount of OsCTF gene expression, which was detected by direct detection, using a Chop-PCR method to detect the methylation level in rice, and real-time quantitative PCR to detect the level of OsCTF gene expression; at the same time,cadmium stress was detected, DNA methylation was analyzed by sulfite sequencing and McrBC-PCR, and OsCTF gene expression was detected by differential expression gene analysis. OsCTF expression and cadmium accumulation in rice after demethylation of the OsCTF 3 ‘- UTR were measured by both direct and cadmium stress assays. Then, OsCTF 3 ‘-UTR demethylation level, OsCTF gene expression and cadmium accumulation in offspring were measured by self-cross method of transgenic rice. Finally, a new rice line with low cadmium accumulation was formed.