Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCR
Abstract Dragon's blood, the red resin derived from the wounded Dracaena, is a precious traditional medicine used by different culture. Dracaena cochinchinensis is one of the main species of Dracaena, and is the endangered medicinal plants in China. The vulnerable status severely limits the med...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-52754-5 |
| _version_ | 1797259340964429824 |
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| author | Shixi Gao Junxiang Peng Mei Rong Yang Liu Yanhong Xu Jianhe Wei |
| author_facet | Shixi Gao Junxiang Peng Mei Rong Yang Liu Yanhong Xu Jianhe Wei |
| author_sort | Shixi Gao |
| collection | DOAJ |
| description | Abstract Dragon's blood, the red resin derived from the wounded Dracaena, is a precious traditional medicine used by different culture. Dracaena cochinchinensis is one of the main species of Dracaena, and is the endangered medicinal plants in China. The vulnerable status severely limits the medicinal value and wide application of dragon’s blood. Therefore, it’s essential to analyze the mechanisms that form dragon’s blood in order to increase artificial production. To clarify the mechanisms forming dragon’s blood, understanding gene expression in the flavonoid biosynthesis pathway is the foundation. However, reference genes of D. cochinchinensis haven’t been analyzed. In this study, expression profiles of seven commonly used housekeeping genes (Actin, α-EF, UBC, β-tubulin, 18S, GAPDH, His) were evaluated by using quantitative real-time PCR combined with the algorithms geNorm, NormFinder, BestKeeper, and RefFinder. On the basis of overall stability ranking, the best reference genes were the combinations β-tubulin +UBC for wounded stems and α-EF +18S + Actin for different organs. Reliability of the recommended reference genes was validated by normalizing relative expression of two key enzyme genes PAL1 and CHI1 in the flavonoid biosynthesis pathway. The results provide a foundation to study gene expression in future research on D. cochinchinensis or other Dracaena. |
| first_indexed | 2024-04-24T23:07:53Z |
| format | Article |
| id | doaj.art-386b6c770dec409992d08f711ae4ee98 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-24T23:07:53Z |
| publishDate | 2024-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-386b6c770dec409992d08f711ae4ee982024-03-17T12:23:51ZengNature PortfolioScientific Reports2045-23222024-03-0114111210.1038/s41598-024-52754-5Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCRShixi Gao0Junxiang Peng1Mei Rong2Yang Liu3Yanhong Xu4Jianhe Wei5Key Laboratory of Bioactive Substances and Rescoures Utilization of Chinese Herbal Medicine, Ministry of Education & National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical CollegeKey Laboratory of Bioactive Substances and Rescoures Utilization of Chinese Herbal Medicine, Ministry of Education & National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical CollegeKey Laboratory of Bioactive Substances and Rescoures Utilization of Chinese Herbal Medicine, Ministry of Education & National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical CollegeKey Laboratory of Bioactive Substances and Rescoures Utilization of Chinese Herbal Medicine, Ministry of Education & National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical CollegeKey Laboratory of Bioactive Substances and Rescoures Utilization of Chinese Herbal Medicine, Ministry of Education & National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical CollegeKey Laboratory of Bioactive Substances and Rescoures Utilization of Chinese Herbal Medicine, Ministry of Education & National Engineering Laboratory for Breeding of Endangered Medicinal Materials, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical CollegeAbstract Dragon's blood, the red resin derived from the wounded Dracaena, is a precious traditional medicine used by different culture. Dracaena cochinchinensis is one of the main species of Dracaena, and is the endangered medicinal plants in China. The vulnerable status severely limits the medicinal value and wide application of dragon’s blood. Therefore, it’s essential to analyze the mechanisms that form dragon’s blood in order to increase artificial production. To clarify the mechanisms forming dragon’s blood, understanding gene expression in the flavonoid biosynthesis pathway is the foundation. However, reference genes of D. cochinchinensis haven’t been analyzed. In this study, expression profiles of seven commonly used housekeeping genes (Actin, α-EF, UBC, β-tubulin, 18S, GAPDH, His) were evaluated by using quantitative real-time PCR combined with the algorithms geNorm, NormFinder, BestKeeper, and RefFinder. On the basis of overall stability ranking, the best reference genes were the combinations β-tubulin +UBC for wounded stems and α-EF +18S + Actin for different organs. Reliability of the recommended reference genes was validated by normalizing relative expression of two key enzyme genes PAL1 and CHI1 in the flavonoid biosynthesis pathway. The results provide a foundation to study gene expression in future research on D. cochinchinensis or other Dracaena.https://doi.org/10.1038/s41598-024-52754-5 |
| spellingShingle | Shixi Gao Junxiang Peng Mei Rong Yang Liu Yanhong Xu Jianhe Wei Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCR Scientific Reports |
| title | Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCR |
| title_full | Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCR |
| title_fullStr | Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCR |
| title_full_unstemmed | Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCR |
| title_short | Screening and validation of reference genes in Dracaena cochinchinensis using quantitative real-time PCR |
| title_sort | screening and validation of reference genes in dracaena cochinchinensis using quantitative real time pcr |
| url | https://doi.org/10.1038/s41598-024-52754-5 |
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