Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica

Background Geraniol 10-hydroxylase (G10H) is a cytochrome P450 monooxygenase involved in regulation, which is involved in the biosynthesis of monoterpene. However, G10H is not characterized at the enzymatic mechanism and regulatory function in Lonicera japonica. Methods and Results A gene related to...

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Main Authors: Shuping Zhang, Zhenhua Liu, Jia Li, Qian Liu, Yongqing Zhang, Gaobin Pu
Format: Article
Language:English
Published: PeerJ Inc. 2025-01-01
Series:PeerJ
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Online Access:https://peerj.com/articles/18832.pdf
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author Shuping Zhang
Zhenhua Liu
Jia Li
Qian Liu
Yongqing Zhang
Gaobin Pu
author_facet Shuping Zhang
Zhenhua Liu
Jia Li
Qian Liu
Yongqing Zhang
Gaobin Pu
author_sort Shuping Zhang
collection DOAJ
description Background Geraniol 10-hydroxylase (G10H) is a cytochrome P450 monooxygenase involved in regulation, which is involved in the biosynthesis of monoterpene. However, G10H is not characterized at the enzymatic mechanism and regulatory function in Lonicera japonica. Methods and Results A gene related to the biosynthesis of monoterpenoid, geraniol 10-hydroxylase, has been cloned from the medicinal plant Lonicera japonica. The gene, LjG10H, encodes a peptide of 498 amino acids with a predicted molecular weight of 54.45 kDa. LjG10H shares a homology of 72.93–83.90% with G10H from other plants. Phylogenetic analysis suggests that the protein encoded by this gene belongs to the cytochrome P450 monooxygenase family. Tissue-specific expression analysis revealed that LjG10H is most highly expressed in flowers. Through heterologous expression in E. coli, the LjG10H protein was purified and its catalytic activity was studied. The results show that the enzyme can catalyze the hydroxylation of geraniol to 10-hydroxygeraniol. Additionally, analysis of Lonicera japonica seedlings with silenced LjG10H revealed a reduction in monoterpenoid content. Conclusions This study demonstrates that LjG10H plays an important role in the biosynthetic pathway of iridoids. This is the first article that ascribes G10H to be associated with the biosynthetic pathway of iridoid. This study provides a theoretical basis for the functional mechanism of LjG10H in regulating iridoid synthesis and provides a valuable resource for molecular breeding studies.
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spelling doaj-art-537a6706820e49a1ae1da70abee9eedd2025-01-15T15:05:30ZengPeerJ Inc.PeerJ2167-83592025-01-0113e1883210.7717/peerj.18832Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonicaShuping ZhangZhenhua LiuJia LiQian LiuYongqing ZhangGaobin PuBackground Geraniol 10-hydroxylase (G10H) is a cytochrome P450 monooxygenase involved in regulation, which is involved in the biosynthesis of monoterpene. However, G10H is not characterized at the enzymatic mechanism and regulatory function in Lonicera japonica. Methods and Results A gene related to the biosynthesis of monoterpenoid, geraniol 10-hydroxylase, has been cloned from the medicinal plant Lonicera japonica. The gene, LjG10H, encodes a peptide of 498 amino acids with a predicted molecular weight of 54.45 kDa. LjG10H shares a homology of 72.93–83.90% with G10H from other plants. Phylogenetic analysis suggests that the protein encoded by this gene belongs to the cytochrome P450 monooxygenase family. Tissue-specific expression analysis revealed that LjG10H is most highly expressed in flowers. Through heterologous expression in E. coli, the LjG10H protein was purified and its catalytic activity was studied. The results show that the enzyme can catalyze the hydroxylation of geraniol to 10-hydroxygeraniol. Additionally, analysis of Lonicera japonica seedlings with silenced LjG10H revealed a reduction in monoterpenoid content. Conclusions This study demonstrates that LjG10H plays an important role in the biosynthetic pathway of iridoids. This is the first article that ascribes G10H to be associated with the biosynthetic pathway of iridoid. This study provides a theoretical basis for the functional mechanism of LjG10H in regulating iridoid synthesis and provides a valuable resource for molecular breeding studies.https://peerj.com/articles/18832.pdfLonicera japonicaIridoidGeraniol-10-hydroxylaseOverexpressionSilencing
spellingShingle Shuping Zhang
Zhenhua Liu
Jia Li
Qian Liu
Yongqing Zhang
Gaobin Pu
Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica
PeerJ
Lonicera japonica
Iridoid
Geraniol-10-hydroxylase
Overexpression
Silencing
title Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica
title_full Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica
title_fullStr Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica
title_full_unstemmed Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica
title_short Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica
title_sort cloning and functional verification of geraniol 10 hydroxylase gene in lonicera japonica
topic Lonicera japonica
Iridoid
Geraniol-10-hydroxylase
Overexpression
Silencing
url https://peerj.com/articles/18832.pdf
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