Targeted metabolite profiling and de novo transcriptome sequencing reveal the key terpene synthase genes in medicinally important plant, Couroupita guianensis Aubl

Hirekodathakallu V. Thulasiram, Shrikant Jagannathrao Karegaonkar, Poojadevi Sharma, Ashish Kumar, Sudha Ramkumar, Avinash Pandreka
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Abstract

The Lecythidaceae family tree, Couroupita guianensis Aubl, popularly known as Nagpushpa, is a widely cultivated ornamental tree with several uses in traditional medicine. The tree is revered as highly sacred in Indian traditional culture due to its uniquely shaped, fragrant flowers. Considering the significance, we were prompted to carry out the metabolite and transcriptome analysis of Nagapushpa. The flower, petals, stamen, stem and leaf of C. guianensis were metabolically profiled, and it was discovered that the flower tissue contained the highest terpenoid reservoir. A number of terpenoid pathway transcripts were also found in the flower tissue after transcriptome profiling. KEGG pathway mapping was carried out to correlate transcript sequences with the biosynthesis of different types of terpenes. We were able to clone three full-length terpene synthase gene candidates, i.e. monoterpene ocimene synthase, diterpene ent-kaurene synthase and sesquiterpene farnesene synthase. The transcript expression of selected terpene synthase genes was also verified in flower tissue. These cloned sequences were used for in silico structural investigations and protein function prediction at the level of 3D structure. The data presented in this study provide a comprehensive resource for the metabolic and transcriptomic profiles of C. guianensis. The study paves the way towards the elucidation of terpene biosynthetic pathway in C. guianensis and heterologous production of useful terpenoids in the future.

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靶向代谢物分析和全新转录组测序揭示了药用植物 Couroupita guianensis Aubl 中的关键萜烯合成酶基因
鹅掌楸科树木 Couroupita guianensis Aubl 俗称 Nagpushpa,是一种广泛栽培的观赏树木,在传统医学中具有多种用途。在印度传统文化中,这种树因其形状独特、芳香四溢的花朵而被尊为圣树。考虑到其重要意义,我们对 Nagapushpa 进行了代谢物和转录组分析。我们对C. guianensis的花、花瓣、雄蕊、茎和叶进行了代谢分析,发现花组织中的萜类化合物储量最高。经过转录组分析,还在花组织中发现了一些萜类化合物通路转录本。我们绘制了 KEGG 通路图,将转录本序列与不同类型萜类化合物的生物合成联系起来。我们克隆了三个全长的萜烯合成酶候选基因,即单萜烯欧西烯合成酶、二萜烯内葵醛合成酶和倍半萜烯法尼烯合成酶。所选萜烯合成酶基因的转录本表达也在花组织中得到了验证。这些克隆序列被用于三维结构层面的硅学结构研究和蛋白质功能预测。本研究提供的数据为 C. guianensis 的代谢和转录组概况提供了全面的资源。该研究为今后阐明滇金丝猴的萜类生物合成途径和异源生产有用的萜类化合物铺平了道路。
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