Molecular Cloning and Functional Characterization of BcTSA in the Biosynthesis of Indole Alkaloids in

Overview

Journal Front Plant Sci

Date 2023 May 4

PMID 37139111

Authors

Zhiying Guo

Junfeng Chen

Zongyou Lv

Yuxiang Huang

Hexin Tan

Lei Zhang

Yong Diao

Affiliations

Soon will be listed here.

Abstract

(Nees) Bremek () is an essential traditional Chinese herb that is commonly used to treat colds, fever, and influenza. Indole alkaloids, such as indigo and indirubin, are the primary active constituents of . The indole-producing reaction is crucial for regulating the flow of indole alkaloids metabolites along the pathways and coordinating primary and secondary product biosynthesis in plants. The tryptophan synthase alpha-subunit (TSA) can catalyse a process that produces indole, which is free to enter secondary metabolite pathways; however, the underlying potential mechanism of regulating indigo alkaloids synthesis remains unknown. Here, a was cloned from the transcriptome of . The BcTSA has a significant degree of similarity with other plant TSAs according to bioinformatics and phylogenetic analyses. Quantitative real-time PCR (RT-qPCR) research showed that was dramatically enhanced in response to treatment with methyl jasmonate (MeJA), salicylic acid (SA), and abscisic acid (ABA), and was predominantly expressed in the stems as opposed to the leaves and rhizomes. Subcellular localization revealed that BcTSA is localized in chloroplasts, which is compatible with the fact that the conversion of indole-3-glycerol phosphate (IGP) to indole occurs in chloroplasts. The complementation assay results showed that BcTSA was functional, demonstrating that it was capable of catalyzing the conversion of IGP to indole. was shown to stimulate the manufacture of indigo alkaloids including isatin, indigo, and indirubin when the gene was overexpressed in the hairy roots of . In conclusion, our research provides novel perspectives that might be applied to manipulating the indole alkaloid composition of .

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References

Liau B, Jong T, Lee M, Chen S . LC-APCI-MS method for detection and analysis of tryptanthrin, indigo, and indirubin in daqingye and banlangen. J Pharm Biomed Anal. 2006; 43(1):346-51. PMC: 7126482. DOI: 10.1016/j.jpba.2006.06.029. View

Maugard T, Enaud E, Choisy P, Legoy M . Identification of an indigo precursor from leaves of Isatis tinctoria (Woad). Phytochemistry. 2001; 58(6):897-904. DOI: 10.1016/s0031-9422(01)00335-1. View

Tan H, Xiao L, Gao S, Li Q, Chen J, Xiao Y . TRICHOME AND ARTEMISININ REGULATOR 1 Is Required for Trichome Development and Artemisinin Biosynthesis in Artemisia annua. Mol Plant. 2015; 8(9):1396-411. DOI: 10.1016/j.molp.2015.04.002. View

Lin W, Huang W, Ning S, Gong X, Ye Q, Wei D . Comparative transcriptome analyses revealed differential strategies of roots and leaves from methyl jasmonate treatment Baphicacanthus cusia (Nees) Bremek and differentially expressed genes involved in tryptophan biosynthesis. PLoS One. 2019; 14(3):e0212863. PMC: 6415880. DOI: 10.1371/journal.pone.0212863. View

Yu J, Zhang Y, Ning S, Ye Q, Tan H, Chen R . Molecular cloning and metabolomic characterization of the 5-enolpyruvylshikimate-3-phosphate synthase gene from Baphicacanthus cusia. BMC Plant Biol. 2019; 19(1):485. PMC: 6842527. DOI: 10.1186/s12870-019-2035-0. View

Xu W, Zhang L, Cunningham A, Li S, Zhuang H, Wang Y . Blue genome: chromosome-scale genome reveals the evolutionary and molecular basis of indigo biosynthesis in Strobilanthes cusia. Plant J. 2020; 104(4):864-879. DOI: 10.1111/tpj.14992. View

Zeng M, Zhong Y, Cai S, Diao Y . Deciphering the bacterial composition in the rhizosphere of Baphicacanthus cusia (NeeS) Bremek. Sci Rep. 2018; 8(1):15831. PMC: 6202335. DOI: 10.1038/s41598-018-34177-1. View

Wang L, Zhou G, Liu P, Song J, Liang Y, Yan X . Dissection of mechanisms of Chinese medicinal formula Realgar-Indigo naturalis as an effective treatment for promyelocytic leukemia. Proc Natl Acad Sci U S A. 2008; 105(12):4826-31. PMC: 2290784. DOI: 10.1073/pnas.0712365105. View

Parr B, Economou C, Herzon S . A concise synthesis of (+)-batzelladine B from simple pyrrole-based starting materials. Nature. 2015; 525(7570):507-10. PMC: 4583359. DOI: 10.1038/nature14902. View

10.

Inoue S, Morita R, Kuwata K, Kunieda T, Ueda H, Hara-Nishimura I . Tissue-specific and intracellular localization of indican synthase from Polygonum tinctorium. Plant Physiol Biochem. 2018; 132:138-144. DOI: 10.1016/j.plaphy.2018.08.034. View

11.

Lee C, Wang C, Hu H, Yen H, Song Y, Yu S . Indole alkaloids indigodoles A-C from aerial parts of Strobilanthes cusia in the traditional Chinese medicine Qing Dai have anti-IL-17 properties. Phytochemistry. 2019; 162:39-46. DOI: 10.1016/j.phytochem.2019.02.016. View

12.

Radwanski E, Last R . Tryptophan biosynthesis and metabolism: biochemical and molecular genetics. Plant Cell. 1995; 7(7):921-34. PMC: 160888. DOI: 10.1105/tpc.7.7.921. View

13.

Feng Q, Zhu G, Gao W, Yang Z, Zhong N, Wang J . Two New Alkaloids from the Roots of Baphicacanthus cusia. Chem Pharm Bull (Tokyo). 2016; 64(10):1505-1508. DOI: 10.1248/cpb.c16-00315. View

14.

Zhu W, Yang B, Komatsu S, Lu X, Li X, Tian J . Binary stress induces an increase in indole alkaloid biosynthesis in Catharanthus roseus. Front Plant Sci. 2015; 6:582. PMC: 4516820. DOI: 10.3389/fpls.2015.00582. View

15.

Huang Y, Tan H, Yu J, Chen Y, Guo Z, Wang G . Stable Internal Reference Genes for Normalizing Real-Time Quantitative PCR in under Hormonal Stimuli and UV Irradiation, and in Different Plant Organs. Front Plant Sci. 2017; 8:668. PMC: 5413499. DOI: 10.3389/fpls.2017.00668. View

16.

Hsu T, Welner D, Russ Z, Cervantes B, Prathuri R, Adams P . Employing a biochemical protecting group for a sustainable indigo dyeing strategy. Nat Chem Biol. 2018; 14(3):256-261. PMC: 5866135. DOI: 10.1038/nchembio.2552. View

17.

Zhang R, Wang B, Ouyang J, Li J, Wang Y . Arabidopsis indole synthase, a homolog of tryptophan synthase alpha, is an enzyme involved in the Trp-independent indole-containing metabolite biosynthesis. J Integr Plant Biol. 2008; 50(9):1070-7. DOI: 10.1111/j.1744-7909.2008.00729.x. View

18.

Kriechbaumer V, Weigang L, Fiesselmann A, Letzel T, Frey M, Gierl A . Characterisation of the tryptophan synthase alpha subunit in maize. BMC Plant Biol. 2008; 8:44. PMC: 2395261. DOI: 10.1186/1471-2229-8-44. View

19.

Hu Y, Ma D, Ning S, Ye Q, Zhao X, Ding Q . High-Quality Genome of the Medicinal Plant Provides Insights Into the Biosynthesis of Indole Alkaloids. Front Plant Sci. 2021; 12:742420. PMC: 8515051. DOI: 10.3389/fpls.2021.742420. View

20.

Salvini M, Boccardi T, Sani E, Bernardi R, Tozzi S, Pugliesi C . Alpha-tryptophan synthase of Isatis tinctoria: gene cloning and expression. Plant Physiol Biochem. 2008; 46(7):715-723. DOI: 10.1016/j.plaphy.2008.04.002. View