TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect ways

Abstract Background Paclitaxel (Taxol) is an invaluable secondary metabolite extracted from Taxus species, wildly utilized in cancer therapeutics. Salicylic acid (SA), an important phytohormone, substantially elevates paclitaxel accumulation in Taxus cell suspension cultures. However, the molecular...

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Main Authors: Yifei Ren, Donglin Liu, Weicheng Zhao, Xinran Wang, Xiaoying Cao, Wen Wan
Format: Article
Language:English
Published: BMC 2025-05-01
Series:BMC Plant Biology
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Online Access:https://doi.org/10.1186/s12870-025-06755-9
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author Yifei Ren
Donglin Liu
Weicheng Zhao
Xinran Wang
Xiaoying Cao
Wen Wan
author_facet Yifei Ren
Donglin Liu
Weicheng Zhao
Xinran Wang
Xiaoying Cao
Wen Wan
author_sort Yifei Ren
collection DOAJ
description Abstract Background Paclitaxel (Taxol) is an invaluable secondary metabolite extracted from Taxus species, wildly utilized in cancer therapeutics. Salicylic acid (SA), an important phytohormone, substantially elevates paclitaxel accumulation in Taxus cell suspension cultures. However, the molecular mechanisms governing SA-induced modulation of paclitaxel biosynthesis remain poorly elucidated. Our previous studies identified TcMYB73, an SA-responsive R2R3-MYB transcription factor (TF), which demonstrates a robust positive correlation with paclitaxel biosynthesis, implying its orchestrating role in this metabolic pathway. Results Expression pattern analysis revealed that TcMYB73 displays predominant expression in lateral roots. Both overexpression and RNA interference (RNAi) of TcMYB73 demonstrated its regulatory function in modulating key paclitaxel biosynthetic genes, including taxadiene synthase (TASY), 10-deacetylbaccatin III-10-O-acetyltransferase (DBAT), and 3’-N-debenzoyl-2’-deoxytaxol-N-benzoyltransferase (DBTNBT). Transient TcMYB73 overexpression in Taxus chinensis (T. chinensis) needles induced 2.38-, 2.87-, and 1.79-fold increases in 10-DAB, baccatin III, and paclitaxel accumulation, respectively, compared to controls. Additionally, yeast one-hybrid (Y1H), Electrophoretic Mobility Shift Assay (EMSA), chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR), and dual-luciferase (Dual-LUC) assays verified that TcMYB73 directly binds to MYB recognition elements in the T10OH promoter, enhancing its transcription. Furthermore, TcWRKY33, a transcriptional activator of DBAT, functions as a positive regulator mediating SA signaling within the paclitaxel biosynthetic pathway. Subsequent investigations validated that TcMYB73 upregulates DBAT expression via direct transcriptional activation of TcWRKY33. Collectively, these results demonstrate that TcMYB73 transduces SA signals to T10OH and TcWRKY33, coordinately regulating paclitaxel biosynthesis through dual mechanisms: direct activation of biosynthetic genes and indirect modulation of upstream regulators. Conclusions Our results indicated that the SA-responsive R2R3-MYB TF, TcMYB73 transcriptionally governs paclitaxel biosynthesis in T. chinensis through direct activation the expression of the T10OH gene, and activating TcWRKY33 expression, thereby modulating DBAT expression. This study provides mechanistic insights into the role of TcMYB73 in mediating SA-induced transcriptional regulation of paclitaxel biosynthesis in Taxus species.
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spelling doaj-art-bfb3e28c29de4bb480e0f770f33646b52025-08-20T03:16:29ZengBMCBMC Plant Biology1471-22292025-05-0125111510.1186/s12870-025-06755-9TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect waysYifei Ren0Donglin Liu1Weicheng Zhao2Xinran Wang3Xiaoying Cao4Wen Wan5School of Life Science, Jiangsu Normal UniversitySchool of Life Science, Jiangsu Normal UniversitySchool of Life Science, Jiangsu Normal UniversitySchool of Life Science, Jiangsu Normal UniversitySchool of Life Science, Jiangsu Normal UniversitySchool of Life Science, Jiangsu Normal UniversityAbstract Background Paclitaxel (Taxol) is an invaluable secondary metabolite extracted from Taxus species, wildly utilized in cancer therapeutics. Salicylic acid (SA), an important phytohormone, substantially elevates paclitaxel accumulation in Taxus cell suspension cultures. However, the molecular mechanisms governing SA-induced modulation of paclitaxel biosynthesis remain poorly elucidated. Our previous studies identified TcMYB73, an SA-responsive R2R3-MYB transcription factor (TF), which demonstrates a robust positive correlation with paclitaxel biosynthesis, implying its orchestrating role in this metabolic pathway. Results Expression pattern analysis revealed that TcMYB73 displays predominant expression in lateral roots. Both overexpression and RNA interference (RNAi) of TcMYB73 demonstrated its regulatory function in modulating key paclitaxel biosynthetic genes, including taxadiene synthase (TASY), 10-deacetylbaccatin III-10-O-acetyltransferase (DBAT), and 3’-N-debenzoyl-2’-deoxytaxol-N-benzoyltransferase (DBTNBT). Transient TcMYB73 overexpression in Taxus chinensis (T. chinensis) needles induced 2.38-, 2.87-, and 1.79-fold increases in 10-DAB, baccatin III, and paclitaxel accumulation, respectively, compared to controls. Additionally, yeast one-hybrid (Y1H), Electrophoretic Mobility Shift Assay (EMSA), chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR), and dual-luciferase (Dual-LUC) assays verified that TcMYB73 directly binds to MYB recognition elements in the T10OH promoter, enhancing its transcription. Furthermore, TcWRKY33, a transcriptional activator of DBAT, functions as a positive regulator mediating SA signaling within the paclitaxel biosynthetic pathway. Subsequent investigations validated that TcMYB73 upregulates DBAT expression via direct transcriptional activation of TcWRKY33. Collectively, these results demonstrate that TcMYB73 transduces SA signals to T10OH and TcWRKY33, coordinately regulating paclitaxel biosynthesis through dual mechanisms: direct activation of biosynthetic genes and indirect modulation of upstream regulators. Conclusions Our results indicated that the SA-responsive R2R3-MYB TF, TcMYB73 transcriptionally governs paclitaxel biosynthesis in T. chinensis through direct activation the expression of the T10OH gene, and activating TcWRKY33 expression, thereby modulating DBAT expression. This study provides mechanistic insights into the role of TcMYB73 in mediating SA-induced transcriptional regulation of paclitaxel biosynthesis in Taxus species.https://doi.org/10.1186/s12870-025-06755-9Taxus chinensisR2R3-MYB transcription factorPaclitaxel biosynthesisSalicylic acid signalTcWRKY33
spellingShingle Yifei Ren
Donglin Liu
Weicheng Zhao
Xinran Wang
Xiaoying Cao
Wen Wan
TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect ways
BMC Plant Biology
Taxus chinensis
R2R3-MYB transcription factor
Paclitaxel biosynthesis
Salicylic acid signal
TcWRKY33
title TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect ways
title_full TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect ways
title_fullStr TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect ways
title_full_unstemmed TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect ways
title_short TcMYB73, a salicylic acid-responsive R2R3-MYB transcription factor, positively regulates paclitaxel biosynthesis in Taxus chinensis in direct and indirect ways
title_sort tcmyb73 a salicylic acid responsive r2r3 myb transcription factor positively regulates paclitaxel biosynthesis in taxus chinensis in direct and indirect ways
topic Taxus chinensis
R2R3-MYB transcription factor
Paclitaxel biosynthesis
Salicylic acid signal
TcWRKY33
url https://doi.org/10.1186/s12870-025-06755-9
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