Huang Yuxiang, Tan Hexin, Li Qing, Wu Xunxun, Guo Zhiying, Chen Junfeng, Zhang Lei, Diao Yong
School of Pharmacy, Quanzhou Medical College, Quanzhou, China.
School of Medicine, Huaqiao University, Quanzhou, China.
Front Plant Sci. 2025 Apr 25;16:1560891. doi: 10.3389/fpls.2025.1560891. eCollection 2025.
(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 gene was isolated and cloned from the transcriptome data of . The full-length cDNA of 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 consists of four exons and three introns. Bioinformatics and phylogenetic analyses revealed a high degree of homology between and its counterparts in various plant species. Quantitative real-time polymerase chain reaction (RT-qPCR) analysis showed that 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 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 protein catalyzed the conversion of shikimic acid to shikimate-3-phosphate. Furthermore, the ectopic overexpression of in significantly enhanced the biosynthetic flux toward indole alkaloids, including indole, indigo, and indirubin. In conclusion, this study identifies and characterizes a novel gene, providing new insights and potential applications for the metabolic engineering of .
（娘家姓）布雷梅克是一种具有药用特性的多年生草本植物，关于其吲哚生物碱生物合成途径中涉及的基因，基因组见解有限。在本研究中，该基因从……的转录组数据中分离并克隆。……的全长cDNA为1657 bp，包括一个265 bp的5'非翻译区、一个507 bp的3'非翻译区和一个编码295个氨基酸的885 bp开放阅读框。……的外显子 - 内含子结构由四个外显子和三个内含子组成。生物信息学和系统发育分析表明，……与其在各种植物物种中的对应物具有高度同源性。定量实时聚合酶链反应（RT - qPCR）分析表明，在包括茉莉酸甲酯（MeJA）、脱落酸（ABA）和紫外线（UV）辐射在内的非生物胁迫条件下，……的表达发生了显著变化。与根、茎和叶相比，该基因在花中主要表达。亚细胞定位分析表明，……主要在叶绿体中表达，证实莽草酸向莽草酸 - 3 - 磷酸的转化发生在该细胞器中。原核表达和酶活性测定表明，异源表达的……蛋白催化了莽草酸向莽草酸 - 3 - 磷酸的转化。此外，……在……中的异位过表达显著增强了包括吲哚、靛蓝和靛玉红在内的吲哚生物碱的生物合成通量。总之，本研究鉴定并表征了一个新的……基因，为……的代谢工程提供了新的见解和潜在应用。
