Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2
Background: Protopanaxatriol (PPT) is an aglycone of ginsenosides, which has high medicinal values. Production of PPT from natural ginseng plants requires artificial deglycosylation procedures of ginsenosides via enzymatic or physicochemical treatments. Metabolic engineering could be an efficient te...
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| פורמט: | Article |
| שפה: | English |
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Elsevier
2019-04-01
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| סדרה: | Journal of Ginseng Research |
| גישה מקוונת: | http://www.sciencedirect.com/science/article/pii/S1226845317303184 |
| _version_ | 1828770061855227904 |
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| author | Yu Shin Gwak Jung Yeon Han Yong Eui Choi |
| author_facet | Yu Shin Gwak Jung Yeon Han Yong Eui Choi |
| author_sort | Yu Shin Gwak |
| collection | DOAJ |
| description | Background: Protopanaxatriol (PPT) is an aglycone of ginsenosides, which has high medicinal values. Production of PPT from natural ginseng plants requires artificial deglycosylation procedures of ginsenosides via enzymatic or physicochemical treatments. Metabolic engineering could be an efficient technology for production of ginsenoside sapogenin. For PPT biosynthesis in Panax ginseng, damarenediol-II synthase (PgDDS) and two cytochrome P450 enzymes (CYP716A47 and CYP716A53v2) are essentially required. Methods: Transgenic tobacco co-overexpressing P. ginseng PgDDS, CYP716A47, and CYP716A53v2 was constructed via Agrobacterium-mediated transformation. Results: Expression of the three introduced genes in transgenic tobacco lines was confirmed by Reverse transcription-polymerase chain reaction (RT-PCR). Analysis of liquid chromatography showed three new peaks, dammarenediol-II (DD), protopanaxadiol (PPD), and PPT, in leaves of transgenic tobacco. Transgenic tobacco (line 6) contained 2.8 μg/g dry weight (DW), 7.3 μg/g DW, and 11.6 μg/g DW of PPT, PPD, and DD in leaves, respectively. Production of PPT was achieved via cell suspension culture and was highly affected by auxin treatment. The content of PPT in cell suspension was increased 37.25-fold compared with that of leaves of the transgenic tobacco. Transgenic tobacco was not able to set seeds because of microspore degeneration in anthers. Transmission electron microscopy analysis revealed that cells of phloem tissue situated in the center of the anther showed an abnormally condensed nuclei and degenerated mitochondria. Conclusion: We successfully achieved the production of PPT in transgenic tobacco. The possible factors deriving male sterility in transgenic tobacco are discussed. Keywords: Male sterility, Metabolic engineering, Protopanaxatriol, Sapogenin, Transgenic tobacco |
| first_indexed | 2024-12-11T14:00:18Z |
| format | Article |
| id | doaj.art-16e8804029a44f0aaedfb37b51a0f75f |
| institution | Directory Open Access Journal |
| issn | 1226-8453 |
| language | English |
| last_indexed | 2024-12-11T14:00:18Z |
| publishDate | 2019-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Ginseng Research |
| spelling | doaj.art-16e8804029a44f0aaedfb37b51a0f75f2022-12-22T01:03:53ZengElsevierJournal of Ginseng Research1226-84532019-04-01432261271Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2Yu Shin Gwak0Jung Yeon Han1Yong Eui Choi2Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon, Republic of KoreaDepartment of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon, Republic of KoreaCorresponding author. Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon 24341, Republic of Korea.; Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon, Republic of KoreaBackground: Protopanaxatriol (PPT) is an aglycone of ginsenosides, which has high medicinal values. Production of PPT from natural ginseng plants requires artificial deglycosylation procedures of ginsenosides via enzymatic or physicochemical treatments. Metabolic engineering could be an efficient technology for production of ginsenoside sapogenin. For PPT biosynthesis in Panax ginseng, damarenediol-II synthase (PgDDS) and two cytochrome P450 enzymes (CYP716A47 and CYP716A53v2) are essentially required. Methods: Transgenic tobacco co-overexpressing P. ginseng PgDDS, CYP716A47, and CYP716A53v2 was constructed via Agrobacterium-mediated transformation. Results: Expression of the three introduced genes in transgenic tobacco lines was confirmed by Reverse transcription-polymerase chain reaction (RT-PCR). Analysis of liquid chromatography showed three new peaks, dammarenediol-II (DD), protopanaxadiol (PPD), and PPT, in leaves of transgenic tobacco. Transgenic tobacco (line 6) contained 2.8 μg/g dry weight (DW), 7.3 μg/g DW, and 11.6 μg/g DW of PPT, PPD, and DD in leaves, respectively. Production of PPT was achieved via cell suspension culture and was highly affected by auxin treatment. The content of PPT in cell suspension was increased 37.25-fold compared with that of leaves of the transgenic tobacco. Transgenic tobacco was not able to set seeds because of microspore degeneration in anthers. Transmission electron microscopy analysis revealed that cells of phloem tissue situated in the center of the anther showed an abnormally condensed nuclei and degenerated mitochondria. Conclusion: We successfully achieved the production of PPT in transgenic tobacco. The possible factors deriving male sterility in transgenic tobacco are discussed. Keywords: Male sterility, Metabolic engineering, Protopanaxatriol, Sapogenin, Transgenic tobaccohttp://www.sciencedirect.com/science/article/pii/S1226845317303184 |
| spellingShingle | Yu Shin Gwak Jung Yeon Han Yong Eui Choi Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2 Journal of Ginseng Research |
| title | Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2 |
| title_full | Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2 |
| title_fullStr | Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2 |
| title_full_unstemmed | Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2 |
| title_short | Production of ginsenoside aglycone (protopanaxatriol) and male sterility of transgenic tobacco co-overexpressing three Panax ginseng genes: PgDDS, CYP716A47, and CYP716A53v2 |
| title_sort | production of ginsenoside aglycone protopanaxatriol and male sterility of transgenic tobacco co overexpressing three panax ginseng genes pgdds cyp716a47 and cyp716a53v2 |
| url | http://www.sciencedirect.com/science/article/pii/S1226845317303184 |
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