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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-05-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/24/9/8430 |
| Summary: | <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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| ISSN: | 1661-6596 1422-0067 |