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| 国槐凋落叶浸提液调控知母甾体皂苷合成的转录组分析 |
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冯超群1,2, 李嘉文1,2, 王晨1,2, 袁云娜1,2, 程子钥1,2, 朱蔚利3, 刘志林4, 刘彩霞4, 刘忠华1,2
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1.北京林业大学 生物科学与技术学院，北京 100083;2.林木育种与生态修复国家工程研究中心，北京 100083;3.榆林市榆阳区农村供水安全中心，陕西 榆林 719000;4.榆林市榆阳区林草生态修复中心，陕西 榆林 719000
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| 摘要: |
| 【目的】探究国槐凋落叶浸提液对知母甾体皂苷活性成分积累的影响，通过转录组测序揭示知母根部 甾体皂苷合成的分子机制。【方法】选取经7.5 g/L国槐凋落叶浸提液处理与蒸馏水处理的知母根部样本，提取总RNA构建cDNA文库，利用Illumina进行转录组测序获得高质量数据，经Trinity拼接组装后得到Unigene；筛选出差异表达基因（DGEs）后进行GO和KEGG富集分析，挖掘知母中与甾体皂苷相关的合成通路及关键酶；选取10个DEGs， 采用RT-qPCR技术进行验证分析。【结果】转录组测序共得到37.25 Gb的高质量数据，拼接组装后获得93 227个单基因族。从7.5 g/L国槐凋落叶浸提液处理组和对照组筛选出4 982个DEGs，其中3 322个DEGs注释到GO数据库的3大分类，1 371个DEGs注释到KEGG数据库的133条代谢通路。与对照组相比，分别有17，3和17个DEGs参与萜类骨架生物合成、半萜和三萜类化合物生物合成、类固醇生物合成。在萜类骨架生物合成途径中，MVK、ISPS、IDI、 HMGCS、HMGCR、FDPS、ACAT均呈现上调表达；在半萜和三萜类化合物生物合成途径中，FDFT1、SQLE均上调表达；在类固醇生物合成途径中，SMO2、SMO1、3BETAHSDD、CYP710A、CYP51、CP11、CAS1、FDFT1、EBP、 SC5DL、TM7SF2、DHCR7均上调表达。RT-qPCR验证结果显示，随机选取的10个DEGs与测序数据趋势一致，进一步证实了转录组结果的可靠性。【结论】国槐凋落叶浸提液处理显著影响知母根部基因表达，并特异性上调了甾体 皂苷生物合成途径中的多个关键酶基因。 |
| 关键词: 知母 国槐凋落叶浸提液 差异表达基因（DEG） 次生代谢产物 合成调控 甾体皂苷合成 |
| DOI：10. 13207/j. jnwafu. 2027. 01. 013 |
| 分类号: |
| 基金项目:国家林业和草原局项目（DNA-2021）；北京市园林绿化局项目（2021-STBHXFC-04-11）；榆林市榆阳区林业局项目（2024XHFW049，2024XHFW059） |
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| Transcriptome analysis on the regulation of steroidal saponin synthesis in Anemarrhena asphodeloides by fallen leafextract of Styphnolobium japonicum |
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Feng Chaoqun1,2, Li Jiawen1,2, Wang Chen1,2, Yuan Yunna1,2, Cheng Ziyue1,2, Zhu Weili3, Liu Zhilin4, Liu Caixia4, Liu Zhonghua1,2
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1.School of Biological Sciences and Technology，Beijing Forestry University，Beijing 100083，China;2.National Engineering Research Center of Tree Breeding and Ecological Restoration，Beijing 100083，China;3.Yuyang District Rural Water Supply Safety Center of Yulin City，Yulin，Shaanxi 719000，China;4.Yuyang District Forest and Grassland Ecological Restoration Center of Yulin City，Yulin，Shaanxi 719000，China
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| Abstract: |
| 【Objective】This study aims to explore the effects of fallen leaf extract from Styphnolobium japonicum（ L.） Schott on the accumulation of active steroidal saponin components in Anemarrhena asphodeloides Bunge，thereby revealing the molecular mechanism of steroidal saponin synthesis in A.asphodeloides roots through transcriptomic sequencing.【Method】Root samples of A.asphodeloides treated with 7.5 g/L fallen leaf extract of S.japonicum and distilled water were collected.Total RNA was extracted for cDNA library construction，and high-quality transcriptomic data were generated via Illumina sequencing. Clean reads were assembled into Unigenes using Trinity. After screening for DGEs，GO and KEGG enrichment analyses were performed to identify synthesis pathways and key enzymes related to steroidal saponins in A.asphodeloides.Ten DEGs were further selected and validated by RT-qPCR.【Result】A total of 37.25 Gb of high-quality data was obtained from transcriptome sequencing，yielding 93 227 Unigenes after assembly.4 982 DEGs were identified from the 7.5 g/L treatment and control groups. Among them，3 322 DEGs were annotated to three major categories in the GO database，and 1 371 DEGs were annotated to 133 metabolic pathways in the KEGG database. Compared with the control group，17，3，and 17 DEGs were involved in the biosynthesis of terpene backbone，sesquiterpe-noids and triterpenoids，and steroids，respectively. In the terpenoid backbone biosynthesis pathway，MVK，ISPS， IDI，HMGCS，HMGCR，FDPS，and ACAT were all upregulated. In the sesquiterpenoid and triterpenoid biosynthesis pathways，FDFT1 and SQLE were both upregulated.In the steroid biosynthesis pathway，SMO2，SMO1，3BETAHSDD，CYP710A，CYP51，CP11，CAS1，FDFT1，EBP，SC5DL，TM7SF2，and DHCR7 were all upregulated. RT-qPCR results confirmed that 10 randomly selected DEGs exhibited trends consistent with sequencing data，further supporting reliability of transcriptome analysis.【Conclusion】Treatment with S.japonica fallen leaf extract significantly influenced gene expression in A.asphodeloides roots，specifically upregulating multiple key enzyme genes in the steroidal saponin biosynthesis pathway. |
| Key words: Anemarrhena asphodeloides fallen leaf extract of Styphnolobium japonicum differentially expressed genes（DEGs） secondary metabolites synthetic regulation steroidal saponin biosynthesis |
