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ORIGINAL RESEARCH article
Front. Plant Sci.
Sec. Plant Biotechnology
Volume 16 - 2025 | doi: 10.3389/fpls.2025.1560891
Molecular cloning and functional characterization of the Shikimate Kinase gene from Baphicacanthus cusia
Provisionally accepted
Yuxiang Huang 1,2,3
Hexin Tan 3*
Qing Li 3
Xunxun Wu 2
Zhiying Guo 2 Junfeng Chen 4
Lei Zhang 3*
Yong Diao 2*- 1 School of Pharmacy, Quanzhou Medical College, Quanzhou, Fujian Province, China
- 2 School of Medicine, Huaqiao University, Quanzhou, Fujian Province, China
- 3 School of Pharmacy, Naval Medical University, Shanghai, China
- 4 Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China
Baphicacanthus cusia (Nee) Bremek, a perennial herbaceous plant with medicinal properties, has limited genomic insights regarding the genes involved in its indole alkaloid biosynthesis pathway. In this study, the BcSK gene was isolated and cloned from the transcriptome data of B. cusia. The full-length cDNA of BcSK is 1,657 bp, comprising a 265 bp 5' UTR, a 507 bp 3' UTR, and an 885 bp ORF encoding 295 amino acids. The exon-intron structure of BcSK consists of four exons and three introns. Bioinformatics and phylogenetic analyses revealed a high degree of homology between BcSK and its counterparts in various plant species.Quantitative real-time polymerase chain reaction (RT-qPCR) analysis showed that BcSK expression was significantly altered under abiotic stress conditions, including methyl jasmonate (MeJA), abscisic acid (ABA), and ultraviolet (UV) radiation. The gene was predominantly expressed in flowers compared to roots, stems, and leaves. Subcellular localization analysis indicated that BcSK is primarily expressed in chloroplasts, confirming that the conversion of shikimic acid to shikimate-3-phosphate occurs in this organelle.Prokaryotic expression and enzyme activity assays demonstrated that the heterologously expressed BcSK protein catalyzed the conversion of shikimic acid to shikimate-3-phosphate.Furthermore, the ectopic overexpression of BcSK in Isatis indigotica significantly enhanced the biosynthetic flux toward indole alkaloids, including indole, indigo, and indirubin.In conclusion, this study identifies and characterizes a novel BcSK gene, providing new insights and potential applications for the metabolic engineering of B. cusia.
Keywords: Baphicacanthus cusia, Shikimate kinase, Indole Alkaloids, Molecular cloning, functional characterization
Received: 15 Jan 2025; Accepted: 17 Mar 2025.
Copyright: © 2025 Huang, Tan, Li, Wu, Guo, Chen, Zhang and Diao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Hexin Tan, School of Pharmacy, Naval Medical University, Shanghai, China
Lei Zhang, School of Pharmacy, Naval Medical University, Shanghai, China
Yong Diao, School of Medicine, Huaqiao University, Quanzhou, 362021, Fujian Province, China
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