Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk.
ObjectiveTo establish Saposhnikovia divaricata (Turcz.) Schischk. hairy root culture system overexpressing p-coumaroyl CoA 2'-hydroxylase (C2'H) gene, and explore the regulatory role of SdC2'H gene in coumarin biosynthesis.MethodThe SdC2'H gene was cloned. A plant binary expressi...
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South China Agricultural University
2025-09-01
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| Series: | Huanan Nongye Daxue xuebao |
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| Online Access: | https://journal.scau.edu.cn/article/doi/10.7671/j.issn.1001-411X.202502001 |
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| author | Wenhui SU Yingchun QI Lichao ZHANG Yi CUI Ruiyun DU Jingjing CUI Yunhe WANG Limin YANG Zhongming HAN |
| author_facet | Wenhui SU Yingchun QI Lichao ZHANG Yi CUI Ruiyun DU Jingjing CUI Yunhe WANG Limin YANG Zhongming HAN |
| author_sort | Wenhui SU |
| collection | DOAJ |
| description | ObjectiveTo establish Saposhnikovia divaricata (Turcz.) Schischk. hairy root culture system overexpressing p-coumaroyl CoA 2'-hydroxylase (C2'H) gene, and explore the regulatory role of SdC2'H gene in coumarin biosynthesis.MethodThe SdC2'H gene was cloned. A plant binary expression vector overexpressing SdC2'H was constructed through gene fusion method. The S. divaricata hairy roots were induced by using Agrobacterium rhizogenes K599 transformation method. The expression level of SdC2'H gene was analyzed by using RT-qPCR. The coumarin content in the hairy roots of S. divaricata was determined by HPLC. ResultThe open reading frame of S. divaricata SdC2'H gene (GenBank accession number: PV169361) was 813 bp in length, and the gene encoded a 270-amino acid protein. The SdC2'H protein was a hydrophilic, unstable protein with no signal peptide and was located outside the membrane. The phylogenetic tree showed that the SdC2'H protein had the closest relationship with Angelica decursiva. In S. divaricata hairy roots overexpressing the SdC2'H gene, the expression level of SdC2'H was significantly elevated compared to blank control (P<0.0001), while the coumarin content markedly exceeded both the blank control (P<0.001) and the empty vector negative control (P<0.01). This indicated that overexpressing SdC2'H gene could improve coumarin content in hairy roots of S. divaricata. ConclusionThis study confirms that SdC2'H gene can promote coumarin biosynthesis in S. divaricata, provides a theoretical basis for analyzing the role of SdC2'H gene in metabolism of coumarins, and lays a foundation for investigating molecular regulatory network of coumarin biosynthesis. |
| format | Article |
| id | doaj-art-6f0b5246ac184b56a78bfb2ae76a9b23 |
| institution | DOAJ |
| issn | 1001-411X |
| language | zho |
| publishDate | 2025-09-01 |
| publisher | South China Agricultural University |
| record_format | Article |
| series | Huanan Nongye Daxue xuebao |
| spelling | doaj-art-6f0b5246ac184b56a78bfb2ae76a9b232025-08-20T03:22:19ZzhoSouth China Agricultural UniversityHuanan Nongye Daxue xuebao1001-411X2025-09-0146575976610.7671/j.issn.1001-411X.202502001202502001Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk.Wenhui SU0Yingchun QI1Lichao ZHANG2Yi CUI3Ruiyun DU4Jingjing CUI5Yunhe WANG6Limin YANG7Zhongming HAN8College of Chinese Medicinal Material, Jilin Agricultural University/Key Laboratory of Ecosystem and Ecological Restoration of Jilin Province, Changchun 130118, ChinaThe 964th Hospital, Changchun 130062, ChinaChengde Chinese Herbal Medicine Green and Ecological Planting Technical Service Center, Chengde 067000, ChinaCollege of Chinese Medicinal Material, Jilin Agricultural University/Key Laboratory of Ecosystem and Ecological Restoration of Jilin Province, Changchun 130118, ChinaCollege of Chinese Medicinal Material, Jilin Agricultural University/Key Laboratory of Ecosystem and Ecological Restoration of Jilin Province, Changchun 130118, ChinaCollege of Chinese Medicinal Material, Jilin Agricultural University/Key Laboratory of Ecosystem and Ecological Restoration of Jilin Province, Changchun 130118, ChinaCollege of Chinese Medicinal Material, Jilin Agricultural University/Key Laboratory of Ecosystem and Ecological Restoration of Jilin Province, Changchun 130118, ChinaCollege of Chinese Medicinal Material, Jilin Agricultural University/Key Laboratory of Ecosystem and Ecological Restoration of Jilin Province, Changchun 130118, ChinaCollege of Chinese Medicinal Material, Jilin Agricultural University/Key Laboratory of Ecosystem and Ecological Restoration of Jilin Province, Changchun 130118, ChinaObjectiveTo establish Saposhnikovia divaricata (Turcz.) Schischk. hairy root culture system overexpressing p-coumaroyl CoA 2'-hydroxylase (C2'H) gene, and explore the regulatory role of SdC2'H gene in coumarin biosynthesis.MethodThe SdC2'H gene was cloned. A plant binary expression vector overexpressing SdC2'H was constructed through gene fusion method. The S. divaricata hairy roots were induced by using Agrobacterium rhizogenes K599 transformation method. The expression level of SdC2'H gene was analyzed by using RT-qPCR. The coumarin content in the hairy roots of S. divaricata was determined by HPLC. ResultThe open reading frame of S. divaricata SdC2'H gene (GenBank accession number: PV169361) was 813 bp in length, and the gene encoded a 270-amino acid protein. The SdC2'H protein was a hydrophilic, unstable protein with no signal peptide and was located outside the membrane. The phylogenetic tree showed that the SdC2'H protein had the closest relationship with Angelica decursiva. In S. divaricata hairy roots overexpressing the SdC2'H gene, the expression level of SdC2'H was significantly elevated compared to blank control (P<0.0001), while the coumarin content markedly exceeded both the blank control (P<0.001) and the empty vector negative control (P<0.01). This indicated that overexpressing SdC2'H gene could improve coumarin content in hairy roots of S. divaricata. ConclusionThis study confirms that SdC2'H gene can promote coumarin biosynthesis in S. divaricata, provides a theoretical basis for analyzing the role of SdC2'H gene in metabolism of coumarins, and lays a foundation for investigating molecular regulatory network of coumarin biosynthesis.https://journal.scau.edu.cn/article/doi/10.7671/j.issn.1001-411X.202502001saposhnikovia divaricata (turcz.) schischk.p-coumaroyl coa 2'-hydroxylase genegene overexpressionhairy rootcoumarin |
| spellingShingle | Wenhui SU Yingchun QI Lichao ZHANG Yi CUI Ruiyun DU Jingjing CUI Yunhe WANG Limin YANG Zhongming HAN Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk. Huanan Nongye Daxue xuebao saposhnikovia divaricata (turcz.) schischk. p-coumaroyl coa 2'-hydroxylase gene gene overexpression hairy root coumarin |
| title | Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk. |
| title_full | Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk. |
| title_fullStr | Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk. |
| title_full_unstemmed | Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk. |
| title_short | Effect of overexpressing SdC2'H gene on coumarin content in hairy root of Saposhnikovia divaricata (Turcz.) Schischk. |
| title_sort | effect of overexpressing sdc2 h gene on coumarin content in hairy root of saposhnikovia divaricata turcz schischk |
| topic | saposhnikovia divaricata (turcz.) schischk. p-coumaroyl coa 2'-hydroxylase gene gene overexpression hairy root coumarin |
| url | https://journal.scau.edu.cn/article/doi/10.7671/j.issn.1001-411X.202502001 |
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