The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i>
<i>Dioscorea zingiberensis</i> is a perennial herb famous for the production of diosgenin, which is a valuable initial material for the industrial synthesis of steroid drugs. Sterol C26-hydroxylases, such as <i>Tf</i>CYP72A616 and <i>Pp</i>CYP72A613, play an impor...
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MDPI AG
2023-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/9/8430 |
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| author | Weipeng Wang Lixiu Hou Song Li Jiaru Li |
| author_facet | Weipeng Wang Lixiu Hou Song Li Jiaru Li |
| author_sort | Weipeng Wang |
| collection | DOAJ |
| description | <i>Dioscorea zingiberensis</i> is a perennial herb famous for the production of diosgenin, which is a valuable initial material for the industrial synthesis of steroid drugs. Sterol C26-hydroxylases, such as <i>Tf</i>CYP72A616 and <i>Pp</i>CYP72A613, play an important role in the diosgenin biosynthesis pathway. In the present study, a novel gene, <i>DzCYP72A12-4</i>, was identified as C26-hydroxylase and was found to be involved in diosgenin biosynthesis, for the first time in <i>D. zingiberensis</i>, using comprehensive methods. Then, the diosgenin heterogenous biosynthesis pathway starting from cholesterol was created in stable transgenic tobacco (<i>Nicotiana tabacum</i> L.) harboring <i>DzCYP90B71</i>(QPZ88854), <i>DzCYP90G6</i>(QPZ88855) and <i>DzCYP72A12-4</i>. Meanwhile, diosgenin was detected in the transgenic tobacco using an ultra-performance liquid chromatography system (Vanquish UPLC 689, Thermo Fisher Scientific, Bremen, Germany) tandem MS (Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer, Thermo Fisher Scientific, Bremen, Germany). Further RT-qPCR analysis showed that <i>DzCYP72A12-4</i> was highly expressed in both rhizomes and leaves and was upregulated under 15% polyethylene glycol (PEG) treatment, indicating that <i>DzCYP72A12-4</i> may be related to drought resistance. In addition, the germination rate of the diosgenin-producing tobacco seeds was higher than that of the negative controls under 15% PEG pressure. In addition, the concentration of malonaldehyde (MDA) was lower in the diosgenin-producing tobacco seedlings than those of the control, indicating higher drought adaptability. The results of this study provide valuable information for further research on diosgenin biosynthesis in <i>D. zingiberensis</i> and its functions related to drought adaptability. |
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| language | English |
| last_indexed | 2024-03-11T04:15:46Z |
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| spelling | doaj.art-fa0336852fa844e3b7b526393f87905d2023-11-17T23:09:31ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-05-01249843010.3390/ijms24098430The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i>Weipeng Wang0Lixiu Hou1Song Li2Jiaru Li3State Key Laboratory of Hybrid Rice, Department of Plant Science, College of Life Sciences, Wuhan University, Wuhan 430072, ChinaState Key Laboratory of Hybrid Rice, Department of Plant Science, College of Life Sciences, Wuhan University, Wuhan 430072, ChinaState Key Laboratory of Hybrid Rice, Department of Plant Science, College of Life Sciences, Wuhan University, Wuhan 430072, ChinaState Key Laboratory of Hybrid Rice, Department of Plant Science, College of Life Sciences, Wuhan University, Wuhan 430072, China<i>Dioscorea zingiberensis</i> is a perennial herb famous for the production of diosgenin, which is a valuable initial material for the industrial synthesis of steroid drugs. Sterol C26-hydroxylases, such as <i>Tf</i>CYP72A616 and <i>Pp</i>CYP72A613, play an important role in the diosgenin biosynthesis pathway. In the present study, a novel gene, <i>DzCYP72A12-4</i>, was identified as C26-hydroxylase and was found to be involved in diosgenin biosynthesis, for the first time in <i>D. zingiberensis</i>, using comprehensive methods. Then, the diosgenin heterogenous biosynthesis pathway starting from cholesterol was created in stable transgenic tobacco (<i>Nicotiana tabacum</i> L.) harboring <i>DzCYP90B71</i>(QPZ88854), <i>DzCYP90G6</i>(QPZ88855) and <i>DzCYP72A12-4</i>. Meanwhile, diosgenin was detected in the transgenic tobacco using an ultra-performance liquid chromatography system (Vanquish UPLC 689, Thermo Fisher Scientific, Bremen, Germany) tandem MS (Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer, Thermo Fisher Scientific, Bremen, Germany). Further RT-qPCR analysis showed that <i>DzCYP72A12-4</i> was highly expressed in both rhizomes and leaves and was upregulated under 15% polyethylene glycol (PEG) treatment, indicating that <i>DzCYP72A12-4</i> may be related to drought resistance. In addition, the germination rate of the diosgenin-producing tobacco seeds was higher than that of the negative controls under 15% PEG pressure. In addition, the concentration of malonaldehyde (MDA) was lower in the diosgenin-producing tobacco seedlings than those of the control, indicating higher drought adaptability. The results of this study provide valuable information for further research on diosgenin biosynthesis in <i>D. zingiberensis</i> and its functions related to drought adaptability.https://www.mdpi.com/1422-0067/24/9/8430<i>Dioscorea zingiberensis</i>diosgeninSterol C26-hydroxylaseCYP72Adrought stress |
| spellingShingle | Weipeng Wang Lixiu Hou Song Li Jiaru Li The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i> International Journal of Molecular Sciences <i>Dioscorea zingiberensis</i> diosgenin Sterol C26-hydroxylase CYP72A drought stress |
| title | The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i> |
| title_full | The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i> |
| title_fullStr | The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i> |
| title_full_unstemmed | The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i> |
| title_short | The Functional Characterization of <i>DzCYP72A12-4</i> Related to Diosgenin Biosynthesis and Drought Adaptability in <i>Dioscorea zingiberensis</i> |
| title_sort | functional characterization of i dzcyp72a12 4 i related to diosgenin biosynthesis and drought adaptability in i dioscorea zingiberensis i |
| topic | <i>Dioscorea zingiberensis</i> diosgenin Sterol C26-hydroxylase CYP72A drought stress |
| url | https://www.mdpi.com/1422-0067/24/9/8430 |
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