Abstract
Glucoraphanin (GRA) produced by the hairy roots of broccoli (Brassica oleracea L. variety Italica Planch) is converted to the secondary metabolite Sulforaphane (SF), which has anticancer properties under specific conditions, and the synthesis of SF is significantly affected by both Ca2+ and Fe2+. Based on the screening of GRA, SF yields, and Epithiospecififier Protein (ESP) activity, the optimal concentration of exogenously added metal ion chelator Ethylene Glycol bis (2-aminoethyl) Tetraacetic Acid (EGTA) for the treatment of broccoli hairy roots and the optimal treatment time were 8.0 mM EGTA for 48 h. Fe2+ content in hairy roots of broccoli reached the lowest level at 48 h, and also significantly reduce the Ca2+ content, but the Ca2+ content recovered with the treatment time and reached the highest level at 48 h. Analysis of transcriptomics data indicated that the physiological and biochemical responses of broccoli hairy roots to exogenous additions of EGTA to metabolic pathways were mainly mediated by the regulation of the expression of relevant genes involved in signal transduction and amino acid metabolism. The expression of NSP2, NSP5, and ESP decreased with the increase of EGTA treatment time and was lowest at 48 h. The expression of 12 genes, such as AOP1.2.7, GSTF11 increased with the increase of EGTA treatment time, and the highest expression was found at 48 h. This study presented a theoretical basis for the study of the exogenous addition of EGTA to alter Ca2+ and Fe2+ contents and affect SF synthesis.
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Data Availability
The sequencing raw data has been uploaded to the Sequence Read Archive (https://www.ncbi.nlm.nih.gov/sra) under Bioproject PRJNA1042919.
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This work was supported by Technology Innovation Guiding Program—Small and Medium Enterprises Innovation Fund (2022–0301-GXC-0192); Gansu Agricultural University Patent Transformation Project (GSAU-JSZR-2021–001); The State Key Laboratory of Aridland Crop Science of China (GSCS-2023–14); and Gansu Province Higher Education Innovation Fund Project (2021B-136).
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Xiaotong Shi designed the experiments and wrote the manuscript. Xiaotong Shi and Jinyu Bao performed the experiments. Xiaotong Shi, Yi Zhao, and Shimin Lan analyzed the data. Sheng Li, Shaoying Ma, Jinyu Bao, and Jie Cao revised the manuscript. All authors read and approved the final manuscript.
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Shi, X., Bao, J., Ma, S. et al. Exogenous addition of EGTA alters Ca2+ and Fe2+ content in broccoli hairy roots and affects sulforaphane production: based on transcriptomic analysis. In Vitro Cell.Dev.Biol.-Plant (2024). https://doi.org/10.1007/s11627-024-10431-2
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DOI: https://doi.org/10.1007/s11627-024-10431-2