Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid Biosynthesis
The medicinal plant <i>Stephania yunnanensis</i> is rich in aporphine alkaloids, a type of benzylisoquinoline alkaloid (BIA), with aporphine being the representative and most abundant compound, but our understanding of the biosynthesis of BIAs in this plant has been relatively limited. P...
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2025-01-01
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| author | Wenlong Shi Qishuang Li Xinyi Li Linglong Luo Jingyi Gan Ying Ma Jian Wang Tong Chen Yifeng Zhang Ping Su Xiaohui Ma Juan Guo Luqi Huang |
| author_facet | Wenlong Shi Qishuang Li Xinyi Li Linglong Luo Jingyi Gan Ying Ma Jian Wang Tong Chen Yifeng Zhang Ping Su Xiaohui Ma Juan Guo Luqi Huang |
| author_sort | Wenlong Shi |
| collection | DOAJ |
| description | The medicinal plant <i>Stephania yunnanensis</i> is rich in aporphine alkaloids, a type of benzylisoquinoline alkaloid (BIA), with aporphine being the representative and most abundant compound, but our understanding of the biosynthesis of BIAs in this plant has been relatively limited. Previous research reported the genome of <i>S. yunnanensis</i> and preliminarily identified the norcoclaurine synthase (NCS), which is involved in the early stages of the BIA biosynthetic pathways. However, the key genes promoting the formation of the aporphine skeleton have not yet been reported. In this study, based on the differences in the content of crebanine and several other BIAs in different tissues, we conducted transcriptome sequencing of roots, stems, and leaves. We then identified candidate genes through functional annotation and sequence alignment and further analyzed them in combination with the genome. Based on this analysis, we identified three CYP80 enzymes (SyCYP80Q5-1, SyCYP80Q5-3, and SyCYP80G6), which exhibited different activities toward (<i>S</i>)- and (<i>R</i>)-configured substrates in <i>S. yunnanensis</i> and demonstrated strict stereoselectivity enroute to aporphine. This study provides metabolomic and transcriptomic information on the biosynthesis of BIAs in <i>S. yunnanensis</i>, offers valuable insights into the elucidation of BIA biosynthesis, and lays the foundation for the complete analysis of pathways for more aporphine alkaloids. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Molecules |
| spelling | doaj-art-48e45f83983b4b31aa48e674e2f0ff2d2025-01-24T13:43:18ZengMDPI AGMolecules1420-30492025-01-0130225910.3390/molecules30020259Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid BiosynthesisWenlong Shi0Qishuang Li1Xinyi Li2Linglong Luo3Jingyi Gan4Ying Ma5Jian Wang6Tong Chen7Yifeng Zhang8Ping Su9Xiaohui Ma10Juan Guo11Luqi Huang12State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaYunnan Key Laboratory of Southern Medicinal Utilization, College of Chinese Materia Medica, Yunnan University of Chinese Medicine, Kunming 650500, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, ChinaThe medicinal plant <i>Stephania yunnanensis</i> is rich in aporphine alkaloids, a type of benzylisoquinoline alkaloid (BIA), with aporphine being the representative and most abundant compound, but our understanding of the biosynthesis of BIAs in this plant has been relatively limited. Previous research reported the genome of <i>S. yunnanensis</i> and preliminarily identified the norcoclaurine synthase (NCS), which is involved in the early stages of the BIA biosynthetic pathways. However, the key genes promoting the formation of the aporphine skeleton have not yet been reported. In this study, based on the differences in the content of crebanine and several other BIAs in different tissues, we conducted transcriptome sequencing of roots, stems, and leaves. We then identified candidate genes through functional annotation and sequence alignment and further analyzed them in combination with the genome. Based on this analysis, we identified three CYP80 enzymes (SyCYP80Q5-1, SyCYP80Q5-3, and SyCYP80G6), which exhibited different activities toward (<i>S</i>)- and (<i>R</i>)-configured substrates in <i>S. yunnanensis</i> and demonstrated strict stereoselectivity enroute to aporphine. This study provides metabolomic and transcriptomic information on the biosynthesis of BIAs in <i>S. yunnanensis</i>, offers valuable insights into the elucidation of BIA biosynthesis, and lays the foundation for the complete analysis of pathways for more aporphine alkaloids.https://www.mdpi.com/1420-3049/30/2/259aporphine alkaloidsbiosynthetic gene clustersbiosynthesisCYP80<i>Stephania yunnanensis</i>transcriptome |
| spellingShingle | Wenlong Shi Qishuang Li Xinyi Li Linglong Luo Jingyi Gan Ying Ma Jian Wang Tong Chen Yifeng Zhang Ping Su Xiaohui Ma Juan Guo Luqi Huang Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid Biosynthesis Molecules aporphine alkaloids biosynthetic gene clusters biosynthesis CYP80 <i>Stephania yunnanensis</i> transcriptome |
| title | Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid Biosynthesis |
| title_full | Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid Biosynthesis |
| title_fullStr | Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid Biosynthesis |
| title_full_unstemmed | Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid Biosynthesis |
| title_short | Transcriptome Analysis of <i>Stephania yunnanensis</i> and Functional Validation of CYP80s Involved in Benzylisoquinoline Alkaloid Biosynthesis |
| title_sort | transcriptome analysis of i stephania yunnanensis i and functional validation of cyp80s involved in benzylisoquinoline alkaloid biosynthesis |
| topic | aporphine alkaloids biosynthetic gene clusters biosynthesis CYP80 <i>Stephania yunnanensis</i> transcriptome |
| url | https://www.mdpi.com/1420-3049/30/2/259 |
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