Wu Shiyi, Lan Kaer, Wang Qian, Su Yi, Li Dengyu, Ma Jing, Hu Tianyuan, Yin Xiaopu, Wei Qiuhui
School of Pharmacy, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China.
School of Pharmacy, Hangzhou Normal University, Hangzhou, Zhejiang, 311121, China; Key Laboratory of Elemene Class Anti-Cancer Chinese Medicines, Engineering Laboratory of Development and Application of Traditional Chinese Medicines, Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou, 311121, China; School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, 200030, China; Xinchang Pharmaceutical Factory, Zhejiang Medicine CO., LTD, China.
Plant Physiol Biochem. 2025 Mar;220:109527. doi: 10.1016/j.plaphy.2025.109527. Epub 2025 Jan 20.
Curcuma wenyujin is acknowledged as a crucial medicinal plant containing essential oils, primarily composed of sesquiterpenoids. While numerous sesquiterpenoids exhibit versatile physiological activities, their levels in C. wenyujin are generally low, particularly the pivotal anti-cancer component elemene. Our previous research demonstrated that basic helix-loop-helix (bHLH) is involved in modulating jasmonate-mediated sesquiterpenoid biosynthesis. In this study, a total of 106 CwbHLHs were identified and systematically analyzed. Under MeJA treatment, the expression levels of CwbHLH15, CwbHLH27, CwbHLH58, CwbHLH73, and CwbHLH89 were significantly upregulated, whereas CwbHLH81 was downregulated. Subsequently, CwbHLH27 was selected for further functional characterization. CwbHLH27 overexpression resulted in increased levels of β-elemene, γ-elemene, β-caryophyllene, and curzerene in C. wenyujin leaves. The expression levels of CwHMGS, CwHMGR, CwDXS, CwDXR, CwFPPS, and CwHDR, key enzyme genes in sesquiterpenoid biosynthesis, were upregulated in transgenic lines. Conversely, CwbHLH27 silencing resulted in the opposite effects. Further analysis revealed that CwbHLH27 activated the transcription of CwHMGS, CwHMGR, and CwDXS by directly binding to the E-box cis-elements within their promoters. Moreover, CwbHLH27 interacts with CwJAZ1/17, thereby executing JA signal transduction and regulating sesquiterpenoid biosynthesis in C. wenyujin. Finally, we elucidated the molecular mechanism by which the CwJAZs-CwbHLH27 regulatory module regulates sesquiterpenoid biosynthesis in response to JA signaling. Our research provides a molecular foundation for biotechnological-assisted breeding of varieties with enhanced active ingredient content.
温郁金被认为是一种重要的药用植物，含有精油，主要由倍半萜类化合物组成。虽然许多倍半萜类化合物具有多种生理活性，但它们在温郁金中的含量普遍较低，尤其是关键的抗癌成分榄香烯。我们之前的研究表明，碱性螺旋-环-螺旋（bHLH）参与调节茉莉酸介导的倍半萜类化合物生物合成。在本研究中，共鉴定并系统分析了106个CwbHLH。在茉莉酸甲酯（MeJA）处理下，CwbHLH15、CwbHLH27、CwbHLH58、CwbHLH73和CwbHLH89的表达水平显著上调，而CwbHLH81则下调。随后，选择CwbHLH27进行进一步的功能表征。CwbHLH27过表达导致温郁金叶片中β-榄香烯、γ-榄香烯、β-石竹烯和莪术烯的含量增加。倍半萜类化合物生物合成中的关键酶基因CwHMGS、CwHMGR、CwDXS、CwDXR、CwFPPS和CwHDR在转基因系中的表达水平上调。相反，CwbHLH27沉默则产生相反的效果。进一步分析表明，CwbHLH27通过直接结合其启动子内的E-box顺式元件来激活CwHMGS、CwHMGR和CwDXS的转录。此外，CwbHLH27与CwJAZ1/17相互作用，从而执行茉莉酸信号转导并调节温郁金中的倍半萜类化合物生物合成。最后，我们阐明了CwJAZs-CwbHLH27调控模块响应茉莉酸信号调节倍半萜类化合物生物合成的分子机制。我们的研究为生物技术辅助培育活性成分含量提高的品种提供了分子基础。
