Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis
Abstract Veratrum (Melanthiaceae; Liliales) is a genus of perennial herbs known for the production of unique bioactive steroidal alkaloids. However, the biosynthesis of these compounds is incompletely understood because many of the downstream enzymatic steps have yet to be resolved. RNA-Seq is a pow...
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| Format: | Article |
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Nature Portfolio
2023-05-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-35429-5 |
| _version_ | 1797822884091002880 |
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| author | Dan Wang Zhijing Yu Meng Guan Qinan Cai Jia Wei Pengda Ma Zheyong Xue Rui Ma Kirsi-Marja Oksman-Caldentey Heiko Rischer |
| author_facet | Dan Wang Zhijing Yu Meng Guan Qinan Cai Jia Wei Pengda Ma Zheyong Xue Rui Ma Kirsi-Marja Oksman-Caldentey Heiko Rischer |
| author_sort | Dan Wang |
| collection | DOAJ |
| description | Abstract Veratrum (Melanthiaceae; Liliales) is a genus of perennial herbs known for the production of unique bioactive steroidal alkaloids. However, the biosynthesis of these compounds is incompletely understood because many of the downstream enzymatic steps have yet to be resolved. RNA-Seq is a powerful method that can be used to identify candidate genes involved in metabolic pathways by comparing the transcriptomes of metabolically active tissues to controls lacking the pathway of interest. The root and leaf transcriptomes of wild Veratrum maackii and Veratrum nigrum plants were sequenced and 437,820 clean reads were assembled into 203,912 unigenes, 47.67% of which were annotated. We identified 235 differentially expressed unigenes potentially involved in the synthesis of steroidal alkaloids. Twenty unigenes, including new candidate cytochrome P450 monooxygenases and transcription factors, were selected for validation by quantitative real-time PCR. Most candidate genes were expressed at higher levels in roots than leaves but showed a consistent profile across both species. Among the 20 unigenes putatively involved in the synthesis of steroidal alkaloids, 14 were already known. We identified three new CYP450 candidates (CYP76A2, CYP76B6 and CYP76AH1) and three new transcription factor candidates (ERF1A, bHLH13 and bHLH66). We propose that ERF1A, CYP90G1-1 and CYP76AH1 are specifically involved in the key steps of steroidal alkaloid biosynthesis in V. maackii roots. Our data represent the first cross-species analysis of steroidal alkaloid biosynthesis in the genus Veratrum and indicate that the metabolic properties of V. maackii and V. nigrum are broadly conserved despite their distinct alkaloid profiles. |
| first_indexed | 2024-03-13T10:15:53Z |
| format | Article |
| id | doaj.art-8c3f7dc478834e9e91b4c7c4e858acea |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-13T10:15:53Z |
| publishDate | 2023-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-8c3f7dc478834e9e91b4c7c4e858acea2023-05-21T11:14:38ZengNature PortfolioScientific Reports2045-23222023-05-0113111410.1038/s41598-023-35429-5Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesisDan Wang0Zhijing Yu1Meng Guan2Qinan Cai3Jia Wei4Pengda Ma5Zheyong Xue6Rui Ma7Kirsi-Marja Oksman-Caldentey8Heiko Rischer9Jilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural SciencesJilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural SciencesKey Laboratory of Saline-Alkali Vegetation Ecology Restoration, Ministry of Education, College of Life Science, Northeast Forestry UniversityJilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural SciencesJilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural SciencesCollege of Life Sciences, Northwest A & F UniversityKey Laboratory of Saline-Alkali Vegetation Ecology Restoration, Ministry of Education, College of Life Science, Northeast Forestry UniversityJilin Provincial Key Laboratory of Agricultural Biotechnology, Jilin Academy of Agricultural SciencesVTT Technical Research Centre of Finland Ltd.VTT Technical Research Centre of Finland Ltd.Abstract Veratrum (Melanthiaceae; Liliales) is a genus of perennial herbs known for the production of unique bioactive steroidal alkaloids. However, the biosynthesis of these compounds is incompletely understood because many of the downstream enzymatic steps have yet to be resolved. RNA-Seq is a powerful method that can be used to identify candidate genes involved in metabolic pathways by comparing the transcriptomes of metabolically active tissues to controls lacking the pathway of interest. The root and leaf transcriptomes of wild Veratrum maackii and Veratrum nigrum plants were sequenced and 437,820 clean reads were assembled into 203,912 unigenes, 47.67% of which were annotated. We identified 235 differentially expressed unigenes potentially involved in the synthesis of steroidal alkaloids. Twenty unigenes, including new candidate cytochrome P450 monooxygenases and transcription factors, were selected for validation by quantitative real-time PCR. Most candidate genes were expressed at higher levels in roots than leaves but showed a consistent profile across both species. Among the 20 unigenes putatively involved in the synthesis of steroidal alkaloids, 14 were already known. We identified three new CYP450 candidates (CYP76A2, CYP76B6 and CYP76AH1) and three new transcription factor candidates (ERF1A, bHLH13 and bHLH66). We propose that ERF1A, CYP90G1-1 and CYP76AH1 are specifically involved in the key steps of steroidal alkaloid biosynthesis in V. maackii roots. Our data represent the first cross-species analysis of steroidal alkaloid biosynthesis in the genus Veratrum and indicate that the metabolic properties of V. maackii and V. nigrum are broadly conserved despite their distinct alkaloid profiles.https://doi.org/10.1038/s41598-023-35429-5 |
| spellingShingle | Dan Wang Zhijing Yu Meng Guan Qinan Cai Jia Wei Pengda Ma Zheyong Xue Rui Ma Kirsi-Marja Oksman-Caldentey Heiko Rischer Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis Scientific Reports |
| title | Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis |
| title_full | Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis |
| title_fullStr | Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis |
| title_full_unstemmed | Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis |
| title_short | Comparative transcriptome analysis of Veratrum maackii and Veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis |
| title_sort | comparative transcriptome analysis of veratrum maackii and veratrum nigrum reveals multiple candidate genes involved in steroidal alkaloid biosynthesis |
| url | https://doi.org/10.1038/s41598-023-35429-5 |
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