Enhancement of Sesquiterpenoid Production by Methyl Jasmonate in Atractylodes chinensis Adventitious Root Culture and its Transcriptional Regulation

Cholil Yun, Solme Pak, Yulin Pang, Jong Min Phyon, Liuyang Yao, Xiaorui Guo, Cholung Kim, Songsu Ma, Tongchol Ri, Ryongho Pak, Huimei Wang
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Abstract

The adventitious root (AR) culture of Atractylodes chinensis is an efficient platform for sustainable production of its sesquiterpenoid compounds (atractylon and β-eudesmol). However, their limited accumulation levels need an effective elicitation approach, and the present study solved this problem using methyl jasmonate (MeJA) as an elicitor. The effects of its treatment concentration and duration on metabolite production were investigated. The ARs treated with 100 µM MeJA for seven days increased atractylon and β-eudesmol by 3.64- and 1.90-fold, respectively, compared with the control. This study further performed transcriptome analysis to explore the transcriptional regulation mechanism of the MeJA elicitation. A total of 124,464 unigenes were identified in A. chinensis ARs, of which 3,568 genes were upregulated and 3,864 genes were downregulated under the MeJA treatment. The MeJA treatment activated the endogenous JA biosynthesis and signaling pathways and sesquiterpenoid biosynthesis. The MeJA treatment more significantly activated the MEP pathway than the MVA pathway. In addition, 14 genes encoding terpene synthase were identified to be significantly upregulated. A total of 2,700 transcription factors (TFs) were identified in A. chinensis ARs, of which Tify, MYB, and MADS were significantly enriched under the MeJA treatment. We predicted a new antagonistic interaction between MYC2 and CPP TFs, which was significantly regulated by the MeJA treatment. The results of real-time quantitative PCR and enzyme activity assays proved the reliability of the transcriptome data. This study will help improve the in vitro production system of A. chinensis sesquiterpenoids and understand the transcriptional regulation mechanism of MeJA elicitation.
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茉莉酸甲酯对白术不定根培养中倍半萜类化合物生产的促进作用及其转录调控
白术的不定根(AR)培养是可持续生产其倍半萜化合物(苍术素和β-桉叶油醇)的有效平台。本研究使用茉莉酸甲酯(MeJA)作为诱导剂解决了这一问题。本研究使用茉莉酸甲酯(MeJA)作为诱导剂,研究了其处理浓度和持续时间对代谢物产生的影响。与对照相比,用 100 µM MeJA 处理 AR 七天后,苍术素和β-桉叶油醇分别增加了 3.64 倍和 1.90 倍。本研究还进一步进行了转录组分析,以探索MeJA诱导的转录调控机制。结果表明,MeJA诱导A. chinensis ARs的转录组共有124,464个基因,其中3,568个基因上调,3,864个基因下调。MeJA处理激活了内源JA生物合成和信号通路以及倍半萜类化合物的生物合成。与 MVA 途径相比,MeJA 处理对 MEP 途径的激活更为显著。此外,还发现有 14 个编码萜烯合成酶的基因被显著上调。在A. chinensis ARs中总共鉴定出2700个转录因子(TFs),其中Tify、MYB和MADS在MeJA处理下明显富集。我们预测 MYC2 与 CPP TFs 之间存在新的拮抗相互作用,MeJA 对其有明显调控作用。实时定量 PCR 和酶活性测定的结果证明了转录组数据的可靠性。本研究将有助于改进A. chinensis倍半萜类化合物的体外生产系统,并了解MeJA诱导的转录调控机制。
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