Isolation and Functional Characterization of Multiple NADPH-Cytochrome P450 Reductase Genes from <i>Camellia sinensis</i> in View of Catechin Biosynthesis

Huang, Ronghao; Liu, Lipeng; He, Xuqiu; Wang, Wenzhao; Hou, Yihong; Chen, Jinfan; Li, Yingying; Zhou, He Ping; Tian, Tian; Wang, Weidong; Xu, Qingshan; Yu, Youben; Zhou, Tong

doi:10.1021/acs.jafc.1c04255

Cited by 4 publications

(10 citation statements)

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“…Both CPR classes were reported to be able to support the in vitro activities of CYPs involved in specialized metabolism ( Rana et al., 2013 ). However, based on numerous functional analyses in planta , CPR class I was believed to be responsible for basal or constitutive metabolisms, while CPR class II was more responsible for adaptation and defense mechanisms, involving numerous specialized metabolisms ( Rana et al., 2013 ; Parage et al., 2016 ; Huang et al., 2021 ; Zou et al., 2021 ). This study showed that LjCPR1 is closely involved with CYP716A51, a C-28 oxidase, which is involved in triterpenoid biosynthesis, one of the specialized metabolisms in L. japonicus .…”

Section: Discussionmentioning

confidence: 99%

“…CPR class I is reported to be constitutively expressed and plays a role in primary or basal constitutive metabolism, while CPR class II is inducible by environmental stimuli and is involved in defense mechanisms through plant secondary metabolism ( Parage et al., 2016 ). Different tissue expression profiles of CPR classes I and II were reported in Withania somnifera ( Rana et al., 2013 ), Panax ginseng ( Zou et al., 2021 ), Camellia sinensis ( Huang et al., 2021 ), and Catharanthus roseus ( Parage et al., 2016 ). The differences in the protein sequences and expression levels of CPR classes I and II suggest that they have different roles in plants.…”

Section: Introductionmentioning

confidence: 99%

“…Transcript and metabolite profiling of stress/elicitor-treated plants or cell cultures can be used to determine gene function in secondary metabolism ( Misra et al., 2014 ). CPR class II genes from W. somnifera , P. ginseng , C. sinensis , and C. roseus were highly induced by methyl jasmonate (MeJA) treatment, whereas their CPR class I genes showed less or no induction ( Rana et al., 2013 ; Parage et al., 2016 ; Huang et al., 2021 ; Zou et al., 2021 ). Interestingly, this different effect of phytohormone treatment was also observed in OSC s and CYP s. RT-PCR analysis of MeJA-treated Ocimum basilicum showed that ObbAS1 and ObCYP2 were significantly and continually induced until 12 h of treatment, while the phytohormone effect on ObbAS2 , ObCYP1 , and ObCYP3 was not that apparent ( Misra et al., 2014 ).…”

Section: Introductionmentioning

confidence: 99%

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Class I and II NADPH-cytochrome P450 reductases exhibit different roles in triterpenoid biosynthesis in Lotus japonicus

et al. 2023

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Cytochrome P450 monooxygenases (CYPs) are enzymes that play critical roles in the structural diversification of triterpenoids. To perform site-specific oxidations of the triterpene scaffold, CYPs require electrons transferred by NADPH-cytochrome P450 reductase (CPR), which is classified into two main classes, class I and class II, based on their structural difference. Lotus japonicus is a triterpenoids-producing model legume with one CPR class I gene (LjCPR1) and a minimum of two CPR class II genes (LjCPR2-1 and LjCPR2-2). CPR classes I and II from different plants have been reported to be involved in different metabolic pathways. By performing gene expression analyses of L. japonicus hairy root culture treated with methyl jasmonate (MeJA), this study revealed that LjCPR1, CYP716A51, and LUS were down-regulated which resulted in no change in betulinic acid and lupeol content. In contrast, LjCPR2s, bAS, CYP93E1, and CYP72A61 were significantly upregulated by MeJA treatment, followed by a significant increase of the precursors for soyasaponins, i.e. β-amyrin, 24-OH β-amyrin, and sophoradiol content. Triterpenoids profile analysis of LORE1 insertion and hairy root mutants showed that the loss of the Ljcpr2-1 gene significantly reduced soyasaponins precursors but not in Ljcpr1 mutants. However, Ljcpr1 and Ljcpr2-1 mutants showed a significant reduction in lupeol and oleanolic, ursolic, and betulinic acid contents. Furthermore, LjCPR1, but not LjCPR2, was crucial for seed development, supporting the previous notion that CPR class I might support plant basal metabolism. This study suggests that CPR classes I and II play different roles in L. japonicus triterpenoid biosynthesis.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Class I and II NADPH-cytochrome P450 reductases exhibit different roles in triterpenoid biosynthesis in Lotus japonicus

et al. 2023

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“…Divergence of pharmacodynamic components in different species/varieties/strains (Qin et al, 2014;, tissues (Guo et al, 2022;Pan et al, 2022) and development stages (Huang et al, 2021;Zhang et al, 2022b) of medicinal plants implicates expression level of key proteins involved in biosynthesis and metabolism of medicinal compounds (Figure 3, yellow part). Based on quantitatively targeted subproteomic analysis of the high and low artemisinin content of sweet wormwood (Artemisia annua), the increase expression of DBR2 accounts for the high artemisinin content .…”

Section: Figurementioning

confidence: 99%

“…Transcript and metabolite profiling of stress/elicitor-treated plants or cell cultures can be used to determine gene function in secondary metabolism . CPR class II genes from W. somnifera, P. ginseng, C. sinensis, and C. roseus were highly induced by methyl jasmonate (MeJA) treatment, whereas their CPR class I genes showed less or no induction (Rana et al, 2013;Parage et al, 2016;Huang et al, 2021;Zou et al, 2021). Interestingly, this different effect of phytohormone treatment was also observed in OSCs and CYPs.…”

Section: Introductionmentioning

confidence: 98%

Advances in Discoveries of Plant Phytochemicals

2024

Frontiers Research Topics

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Frontiers is more than just an open access publisher of scholarly articles: it is a pioneering approach to the world of academia, radically improving the way scholarly research is managed. The grand vision of Frontiers is a world where all people have an equal opportunity to seek, share and generate knowledge. Frontiers provides immediate and permanent online open access to all its publications, but this alone is not enough to realize our grand goals. Frontiers journal seriesThe Frontiers journal series is a multi-tier and interdisciplinary set of openaccess, online journals, promising a paradigm shift from the current review, selection and dissemination processes in academic publishing. All Frontiers journals are driven by researchers for researchers; therefore, they constitute a service to the scholarly community. At the same time, the Frontiers journal series operates on a revolutionary invention, the tiered publishing system, initially addressing specific communities of scholars, and gradually climbing up to broader public understanding, thus serving the interests of the lay society, too. Dedication to qualityEach Frontiers article is a landmark of the highest quality, thanks to genuinely collaborative interactions between authors and review editors, who include some of the world's best academicians. Research must be certified by peers before entering a stream of knowledge that may eventually reach the public -and shape society; therefore, Frontiers only applies the most rigorous and unbiased reviews. Frontiers revolutionizes research publishing by freely delivering the most outstanding research, evaluated with no bias from both the academic and social point of view. By applying the most advanced information technologies, Frontiers is catapulting scholarly publishing into a new generation. What are Frontiers Research Topics?Frontiers Research Topics are very popular trademarks of the Frontiers journals series: they are collections of at least ten articles, all centered on a particular subject. With their unique mix of varied contributions from Original Research to Review Articles, Frontiers Research Topics unify the most influential researchers, the latest key findings and historical advances in a hot research area.

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A flavonoid metabolon: cytochrome b₅ enhances B‐ring trihydroxylated flavan‐3‐ols synthesis in tea plants

Ruan,

Gao,

Fang

et al. 2024

The Plant Journal

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SUMMARYFlavan‐3‐ols are prominent phenolic compounds found abundantly in the young leaves of tea plants. The enzymes involved in flavan‐3‐ol biosynthesis in tea plants have been extensively investigated. However, the localization and associations of these numerous functional enzymes within cells have been largely neglected. In this study, we aimed to investigate the synthesis of flavan‐3‐ols in tea plants, particularly focusing on epigallocatechin gallate. Our analysis involving the DESI‐MSI method to reveal a distinct distribution pattern of B‐ring trihydroxylated flavonoids, primarily concentrated in the outer layer of buds. Subcellular localization showed that CsC4H, CsF3′H, and CsF3′5′H localizes endoplasmic reticulum. Protein–protein interaction studies demonstrated direct associations between CsC4H, CsF3′H, and cytoplasmic enzymes (CHS, CHI, F3H, DFR, FLS, and ANR), highlighting their interactions within the biosynthetic pathway. Notably, CsF3′5′H, the enzyme for B‐ring trihydroxylation, did not directly interact with other enzymes. We identified cytochrome b5 isoform C serving as an essential redox partner, ensuring the proper functioning of CsF3′5′H. Our findings suggest the existence of distinct modules governing the synthesis of different B‐ring hydroxylation compounds. This study provides valuable insights into the mechanisms underlying flavonoid diversity and efficient synthesis and enhances our understanding of the substantial accumulation of B‐ring trihydroxylated flavan‐3‐ols in tea plants.

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Isolation and Functional Characterization of Multiple NADPH-Cytochrome P450 Reductase Genes from Camellia sinensis in View of Catechin Biosynthesis

Cited by 4 publications

References 48 publications

Class I and II NADPH-cytochrome P450 reductases exhibit different roles in triterpenoid biosynthesis in Lotus japonicus

Class I and II NADPH-cytochrome P450 reductases exhibit different roles in triterpenoid biosynthesis in Lotus japonicus

Advances in Discoveries of Plant Phytochemicals

A flavonoid metabolon: cytochrome b₅ enhances B‐ring trihydroxylated flavan‐3‐ols synthesis in tea plants

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Isolation and Functional Characterization of Multiple NADPH-Cytochrome P450 Reductase Genes from Camellia sinensis in View of Catechin Biosynthesis

Cited by 4 publications

References 48 publications

Class I and II NADPH-cytochrome P450 reductases exhibit different roles in triterpenoid biosynthesis in Lotus japonicus

Class I and II NADPH-cytochrome P450 reductases exhibit different roles in triterpenoid biosynthesis in Lotus japonicus

Advances in Discoveries of Plant Phytochemicals

A flavonoid metabolon: cytochrome b5 enhances B‐ring trihydroxylated flavan‐3‐ols synthesis in tea plants

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A flavonoid metabolon: cytochrome b₅ enhances B‐ring trihydroxylated flavan‐3‐ols synthesis in tea plants