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OBJECTIVE To investigate the biosynthetic pathway of C21 steroidal compounds in Cynanchum otophyllum Schneid. METHODS Metabolomics and transcriptomics were used to compare and analyze the relative contents of C21 steroids in the roots, stems and leaves of C. otophyllum. Then, the genes related to C21 steroid biosynthesis in C. otophyllum were screened through Kyoto Encyclopedia of Genes and Genomes(KEGG) database annotation. Finally, some differentially expressed genes (DEGs) were verified by quantitative real-time PCR. RESULTS The qingyangshengenin content in the roots was significantly up-regulated, with the relative qingyangshengenin content in the roots being approximately 73.10 times higher than that in the leaves and 19.05 times higher than that in the stems. Transcriptomic analysis revealed that 269 DEGs annotated C21 steroidal biosynthetic pathways. By analyzing the DEGs annotated between the comparison groups, 18 key enzymes were screened out in the C21 steroidal synthesis pathway, which were encoded by 87 genes, among which AACT and other enzymes were the key enzymes in the upstream stage of the biosynthesis pathway. Quantitative real-time PCR was performed to verify the expression trend of eight DEGs, which was consistent with the corresponding transcriptome data. CONCLUSION The biosynthetic pathway of C21 steroidal compounds is systematically analyzed in this study, and several key enzymes and coding genes are screened, which enrich the omics data of C. otophyllum. These findings lay a foundation for further study on the biosynthesis mechanism of C21 steroid compounds of C. otophyllum.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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