Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i>
<i>Uncaria rhynchophylla</i>, a medicinal plant extensively used in traditional Chinese medicine, is an important plant source of terpenoid indole alkaloids (TIAs), but the mechanism of TIA biosynthesis at molecular level remains unclear. Geraniol synthase (GES) serves as a crucial enzym...
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2025-07-01
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| author | Xinghui Liu Wenqiang Chen Linxuan Li Detian Mu Iain W. Wilson Xueshuang Huang Yahui Xiang Lina Zhu Limei Pan Deyou Qiu Qi Tang |
| author_facet | Xinghui Liu Wenqiang Chen Linxuan Li Detian Mu Iain W. Wilson Xueshuang Huang Yahui Xiang Lina Zhu Limei Pan Deyou Qiu Qi Tang |
| author_sort | Xinghui Liu |
| collection | DOAJ |
| description | <i>Uncaria rhynchophylla</i>, a medicinal plant extensively used in traditional Chinese medicine, is an important plant source of terpenoid indole alkaloids (TIAs), but the mechanism of TIA biosynthesis at molecular level remains unclear. Geraniol synthase (GES) serves as a crucial enzyme in catalyzing the formation of geraniol from geranyl pyrophosphate (GPP) in various plants, but the functional characterization of the <i>GES</i> gene in <i>U. rhynchophylla</i> has not been investigated. In this study, a <i>GES</i> was identified and characterized through genome mining and bioinformatic analysis. Functional validation was performed via a protein catalysis experiment, transient expression in <i>Nicotiana benthamiana</i>, and methyl jasmonate (MeJA) induction experiments. The full-length <i>UrGES</i> gene was 1761 bp, encoding a protein product of 586 amino acids with an estimated 67.5 kDa molecular weight. Multiple sequence alignments and phylogenetic analysis placed UrGES within the terpene synthase g (TPS-g) subfamily, showing high similarity to known GESs from other plants. Enzymatic assays confirmed that recombinant UrGES catalyzed GPP conversion to a single product of geraniol. The transient expression of <i>UrGES</i> resulted in geraniol accumulation in <i>N. benthamiana</i>, further confirming its function in vivo. <i>UrGES</i> expression was observed in leaves, stems, and roots, where leaves had the highest transcript levels. Moreover, MeJA treatment significantly upregulated <i>UrGES</i> expression, which positively correlated with an increase in alkaloid content. This study functionally characterizes UrGES as a geraniol synthase in <i>U. rhynchophylla</i>, contributing to the current knowledge of the TIA biosynthetic pathway. These findings may offer insights for future metabolic engineering aiming to enhance TIA yields for pharmaceutical and industrial applications. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-07-01 |
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| spelling | doaj-art-d5deb16a31f64845b4f307f8fa7af95f2025-08-20T04:00:55ZengMDPI AGPlants2223-77472025-07-011415227310.3390/plants14152273Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i>Xinghui Liu0Wenqiang Chen1Linxuan Li2Detian Mu3Iain W. Wilson4Xueshuang Huang5Yahui Xiang6Lina Zhu7Limei Pan8Deyou Qiu9Qi Tang10National Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, College of Horticulture, Hunan Agricultural University, Changsha 410128, ChinaNational Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, College of Horticulture, Hunan Agricultural University, Changsha 410128, ChinaGuangxi Key Laboratory of High-Quality Formation and Utilization of Dao-di Herbs, National Center for TCM Inheritance and Innovation, Guangxi Botanical Garden of Medicinal Plants, Nanning 530023, ChinaNational Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, College of Horticulture, Hunan Agricultural University, Changsha 410128, ChinaCSIRO Agriculture and Food, Canberra, ACT 2601, AustraliaHunan Provincial Key Laboratory for Synthetic Biology of Traditional Chinese Medicine, Hunan University of Medicine, Huaihua 418000, ChinaNational Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, College of Horticulture, Hunan Agricultural University, Changsha 410128, ChinaNational Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, College of Horticulture, Hunan Agricultural University, Changsha 410128, ChinaGuangxi Key Laboratory of High-Quality Formation and Utilization of Dao-di Herbs, National Center for TCM Inheritance and Innovation, Guangxi Botanical Garden of Medicinal Plants, Nanning 530023, ChinaState Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, ChinaNational Research Center of Engineering Technology for Utilization of Botanical Functional Ingredients, College of Horticulture, Hunan Agricultural University, Changsha 410128, China<i>Uncaria rhynchophylla</i>, a medicinal plant extensively used in traditional Chinese medicine, is an important plant source of terpenoid indole alkaloids (TIAs), but the mechanism of TIA biosynthesis at molecular level remains unclear. Geraniol synthase (GES) serves as a crucial enzyme in catalyzing the formation of geraniol from geranyl pyrophosphate (GPP) in various plants, but the functional characterization of the <i>GES</i> gene in <i>U. rhynchophylla</i> has not been investigated. In this study, a <i>GES</i> was identified and characterized through genome mining and bioinformatic analysis. Functional validation was performed via a protein catalysis experiment, transient expression in <i>Nicotiana benthamiana</i>, and methyl jasmonate (MeJA) induction experiments. The full-length <i>UrGES</i> gene was 1761 bp, encoding a protein product of 586 amino acids with an estimated 67.5 kDa molecular weight. Multiple sequence alignments and phylogenetic analysis placed UrGES within the terpene synthase g (TPS-g) subfamily, showing high similarity to known GESs from other plants. Enzymatic assays confirmed that recombinant UrGES catalyzed GPP conversion to a single product of geraniol. The transient expression of <i>UrGES</i> resulted in geraniol accumulation in <i>N. benthamiana</i>, further confirming its function in vivo. <i>UrGES</i> expression was observed in leaves, stems, and roots, where leaves had the highest transcript levels. Moreover, MeJA treatment significantly upregulated <i>UrGES</i> expression, which positively correlated with an increase in alkaloid content. This study functionally characterizes UrGES as a geraniol synthase in <i>U. rhynchophylla</i>, contributing to the current knowledge of the TIA biosynthetic pathway. These findings may offer insights for future metabolic engineering aiming to enhance TIA yields for pharmaceutical and industrial applications.https://www.mdpi.com/2223-7747/14/15/2273terpene synthasebioinformatics analysisenzymatic assaytransient expressionterpenoid indole alkaloids |
| spellingShingle | Xinghui Liu Wenqiang Chen Linxuan Li Detian Mu Iain W. Wilson Xueshuang Huang Yahui Xiang Lina Zhu Limei Pan Deyou Qiu Qi Tang Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i> Plants terpene synthase bioinformatics analysis enzymatic assay transient expression terpenoid indole alkaloids |
| title | Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i> |
| title_full | Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i> |
| title_fullStr | Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i> |
| title_full_unstemmed | Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i> |
| title_short | Identification and Functional Characterization of a Geraniol Synthase UrGES from <i>Uncaria rhynchophylla</i> |
| title_sort | identification and functional characterization of a geraniol synthase urges from i uncaria rhynchophylla i |
| topic | terpene synthase bioinformatics analysis enzymatic assay transient expression terpenoid indole alkaloids |
| url | https://www.mdpi.com/2223-7747/14/15/2273 |
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