Metabolic engineering of the anthocyanin biosynthetic pathway in Artemisia annua and relation to the expression of the artemisinin biosynthetic pathway

Judd, Rika; Dong, Yilun; Sun, Xiaomei; Zhu, Yue; Li, Mingzhuo; Xie, De‐Yu

doi:10.1007/s00425-023-04091-6

Cited by 3 publications

(9 citation statements)

References 69 publications

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“…We recently reported our methods and steps for medium preparation and subculture of red cells (Judd et al 2023). Herein, we followed our protocols reported.…”

Section: Methodsmentioning

confidence: 99%

“…We previously reported 17 anthocyanin peaks detected in TAPA cells (Judd et al 2023). To understand whether 2, 4-D could alter the profiles of anthocyanins, HPLC-qTOF-MS/MS was performed.…”

Section: Effects Of 24-d On Anthocyanin Profilesmentioning

confidence: 98%

“…The extraction steps followed the manufacturer's instructions. The extracted RNA samples were treated with DNase as described previously (Judd et al 2023). The resulting DNA-free RNA samples were stored in a -80 freezer.…”

Section: Real Time Reverse Transcription-polymerase Chain Reactionmentioning

confidence: 99%

“…The profiles of anthocyanins were analyzed on Agilent 6210 time-of-flight LC/MS (Agilent Technologies, Santa Clara, CA, USA) as described previously (Judd et al 2023). In brief, the HPLC mobile phase solvents consisted of 1% acetic acid in LC-MS grade water (A) and 100% acetonitrile (B).…”

Section: Hplc-qtof-ms Analysis Of Anthocyaninsmentioning

confidence: 99%

“…Although no anthocyanin structures were reported from A. annua in the past (Brown 2010), we recently used Production of Anthocyanin Pigment 1 (PAP1), a master MYB regulator of Arabidopsis anthocyanin biosynthesis, to have established red cell cultures from this antimalarial plant (Judd et al 2023). We detected 17 anthocyanin molecules from the extracts of red cells.…”

Section: Introductionmentioning

confidence: 99%

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Molecular understanding of anthocyanin biosynthesis activated by PAP1 in engineered redArtemisia annuacells and regulation of 2, 4-dichlorophenoxyacetic acid

Dong

Cruz

et al. 2023

Preprint

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Artemisia annua is an effective antimalarial medicinal crop. We have established anthocyanin-producing red cell cultures from this plant with the overexpression of Production of Anthocyanin Pigment 1 (PAP1) encoding a R2R3MYB transcription factor. To understand the molecular mechanism by which PAP1 activated the entire anthocyanin pathway, we mined the genomic sequences of A. annua and obtained eight promoters of the anthocyanin pathway genes. Sequence analysis identified four types of AC cis-elements from six promoters, the MYB response elements (MRE) bound by PAP1. In addition, six promoters were determined to have at least one G-Box cis-element. Eight promoters were cloned for activity analysis. Duel luciferase assays showed that PAP1 significantly enhanced the promoting activity of seven promoters, indicating that PAP1 turned on the biosynthesis of anthocyanins via the activation of these pathway gene expression. To understand how 2,4-dichlorophenoxyacetic acid (2,4-D), an auxin, regulates the PAP1-activated anthocyanin biosynthesis, five different concentrations (0, 0.05, 0.5, 2.5, and 5 micro M) were tested to characterize anthocyanin production and profiles. The resulting data showed that the concentrations tested decreased the fresh weight of callus growth, anthocyanin levels, and the production of anthocyanins per petri dish. HPLC-qTOF-MS/MS based profiling showed that these concentrations did not alter anthocyanin profiles. Real time RT-PCR was completed to characterize the expression PAP1 and four representative pathway genes. The results showed that the five concentrations reduced the expression levels of the constitutive PAP1 transgene and three pathway genes significantly and eliminated chalcone synthase gene in expression either significantly or slightly. These data indicate that the constitutive PAP1 expression depends on gradients added in the medium. Based on these findings, the regulation of 2,4-D is discussed for anthocyanin engineering in red cells of A. annua.

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“…We recently reported our methods and steps for medium preparation and subculture of red cells (Judd et al 2023). Herein, we followed our protocols reported.…”

Section: Methodsmentioning

confidence: 99%

“…We previously reported 17 anthocyanin peaks detected in TAPA cells (Judd et al 2023). To understand whether 2, 4-D could alter the profiles of anthocyanins, HPLC-qTOF-MS/MS was performed.…”

Section: Effects Of 24-d On Anthocyanin Profilesmentioning

confidence: 98%

Section: Real Time Reverse Transcription-polymerase Chain Reactionmentioning

confidence: 99%

Section: Hplc-qtof-ms Analysis Of Anthocyaninsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Molecular understanding of anthocyanin biosynthesis activated by PAP1 in engineered redArtemisia annuacells and regulation of 2, 4-dichlorophenoxyacetic acid

Dong

Cruz

et al. 2023

Preprint

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Molecular understanding of anthocyanin biosynthesis activated by PAP1 and regulated by 2, 4-dichlorophenoxyacetic acid in engineered red Artemisia annua cells

Dong,

Li,

Cruz

et al. 2023

Planta

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Metabolic engineering of microbial and plant chasses to optimize artemisinin production

Behera,

Ekwudo,

Nohoesu

et al. 2023

CABI Reviews

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Artemisinin, a sesquiterpene lactone extracted from the sweet wormwood plant ( Artemisia annua ), is a crucial component in the production of artemisinin-based combination therapies (ACTs), the most effective treatment for malaria. In recent years, metabolic engineering has emerged as a powerful tool to enhance the production of artemisinin, both in microbial and plant chasses. This review article provides an overview of the recent literature on the metabolic engineering strategies employed to optimize artemisinin production, highlighting breakthroughs, challenges, and future directions.

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Metabolic engineering of the anthocyanin biosynthetic pathway in Artemisia annua and relation to the expression of the artemisinin biosynthetic pathway

Cited by 3 publications

References 69 publications

Molecular understanding of anthocyanin biosynthesis activated by PAP1 in engineered redArtemisia annuacells and regulation of 2, 4-dichlorophenoxyacetic acid

Molecular understanding of anthocyanin biosynthesis activated by PAP1 in engineered redArtemisia annuacells and regulation of 2, 4-dichlorophenoxyacetic acid

Molecular understanding of anthocyanin biosynthesis activated by PAP1 and regulated by 2, 4-dichlorophenoxyacetic acid in engineered red Artemisia annua cells

Metabolic engineering of microbial and plant chasses to optimize artemisinin production

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