Bano Anjum, Kumari Archana, Pandey Akansha, Kumar Akhilesh, Madan Mallika, Mohanta Anshu, Minj Emma Anjali, Pandey Tushar, Kanojiya Sanjeev, Pandey Richa, Shukla Rakesh Kumar, Tripathi Vineeta
Botany Unit (SAIF & R), CSIR-CDRI, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226031, Uttar Pradesh, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India.
Protoplasma. 2025 Jan 23. doi: 10.1007/s00709-025-02030-8.
Eclipta prostrata belongs to the Asteraceae family. The plant contains bioactive compounds like wedelolactone (WDL) and demethylwedelolactone (DW). Its transcriptomic information engaged with secondary metabolite biosynthesis is not available. Based on differential accumulation of WDL and DW in root, shoot of the mature plant, we performed comparative de novo transcriptome of root and shoot tissue in three independent biological replicates and generated 49820 unique transcripts. Annotation resulted in significant matches for 43,015 unigenes. Based on differential gene expression data, we found WDL biosynthesis-related transcripts, which were mainly upregulated in shoot. Finally, 13 selected differentially expressed transcripts related to WDL biosynthesis that were validated by qRT-PCR. Detailed tissue-specific metabolite and transcript profiling revealed that DW highly accumulated in root and WDL accumulation was high in aerial part along with transcripts. For WDL pathway exploration, we did integrated profiling of 08 metabolites and 13 transcripts and witnessed that only naringenin, apigenin, DW, and WDL were detected in different developmental stages. Taking leads from the findings, we postulated that naringenin to apigenin pathway is one potential route for WDL biosynthesis. Moreover, wound stress led to accumulation of DW and WDL and related biosynthetic transcripts. Furthermore, the selected enzymes were subjected to molecular docking and binding studies for the predicted substrates involved in crucial and advance steps of WDL biosynthesis. A comprehensive analysis integrating de novo transcriptomics, metabolomics, and molecular docking of targeted proteins paves the way for the elucidation of the putative wedelolactone biosynthesis pathway from E. prostrata.
鳢肠属于菊科。该植物含有如去甲氧基淡黄木樨草素（WDL）和去甲基去甲氧基淡黄木樨草素（DW）等生物活性化合物。其与次生代谢物生物合成相关的转录组信息尚未可知。基于成熟植株根和地上部分中WDL和DW的差异积累，我们对根和地上部分组织进行了三次独立生物学重复的比较从头转录组分析，并生成了49820个独特转录本。注释结果显示43015个单基因有显著匹配。基于差异基因表达数据，我们发现了与WDL生物合成相关的转录本，这些转录本主要在地上部分上调。最后，通过qRT-PCR验证了13个与WDL生物合成相关的差异表达转录本。详细的组织特异性代谢物和转录本分析表明，DW在根中高度积累，而WDL在地上部分与转录本一起积累量较高。为了探索WDL途径，我们对8种代谢物和13个转录本进行了综合分析，发现在不同发育阶段仅检测到柚皮素、芹菜素、DW和WDL。根据这些发现，我们推测柚皮素到芹菜素的途径是WDL生物合成的一条潜在途径。此外，伤口胁迫导致DW、WDL和相关生物合成转录本的积累。此外，对所选酶进行了分子对接和与WDL生物合成关键和进阶步骤中涉及的预测底物的结合研究。整合从头转录组学、代谢组学和靶向蛋白质分子对接的综合分析为阐明鳢肠中假定的去甲氧基淡黄木樨草素生物合成途径铺平了道路。
