Yang Yi, Zhang Cui-Ping, Liu Xing, Wei Yue, Wu Neng-Biao
School of Life Science, Southwest University, Chongqing 400715, China.
Key Laboratory of Eco-environments in Three Gorges Region, Ministry of Education, Chongqing 400715, China.
Zhongguo Zhong Yao Za Zhi. 2018 Oct;43(20):4044-4049. doi: 10.19540/j.cnki.cjcmm.20180726.018.
Hyoscyamine and scopolamine are important secondary metabolites of tropane alkaloid in Atropa belladonna with pharmacological values in many aspects.In this study, the seedlings of A.belladonna were planted by soil culture and treated with different concentrations of methyl jasmonate (MeJA). The contents of hyoscyamine and scopolamine,the upstream products in alkaloid synthesis,and the expression levels of key enzyme genes PMT, TR Ⅰ and H6H in secondary metabolites of A. belladonna seedlings were measured to clarify the mechanism of MeJA regulating alkaloids synthesis.The results showed that MeJA(200 μmol·L⁻¹) treatment was more favorable for the accumulation of alkaloids.The content of putrescine was almost consistent with the change of key enzymes activities in the synthesis of putrescine,the both increased first and then decreased with the increased MeJA concentration and the content of putrescine reached the highest at 200 μmol·L⁻¹ MeJA.Further detection of gene expression of PMT, TR Ⅰ and H6H in TAs synthesis pathway showed that no significant trend in PMT gene expression levels.The expression levels of TR Ⅰ and H6H in leaves and roots under 200 μmol·L⁻¹ MeJA were the highest.It can be speculated that the regulation of the formation of hyoscyamine and scopolamine by MeJA mainly through affecting the expression of key enzyme genes.Appropriate concentration of MeJA increased the gene expression of TR Ⅰ in both leaves and roots as well as H6H in roots,promoting the accumulation of alkaloids and the conversion of hyoscyamine to scopolamine.
莨菪碱和东莨菪碱是颠茄中托烷生物碱的重要次生代谢产物,在许多方面具有药理价值。在本研究中,通过土壤培养种植颠茄幼苗,并用不同浓度的茉莉酸甲酯(MeJA)进行处理。测定了颠茄幼苗生物碱合成上游产物莨菪碱和东莨菪碱的含量,以及次生代谢产物中关键酶基因PMT、TRⅠ和H6H的表达水平,以阐明MeJA调节生物碱合成的机制。结果表明,MeJA(200μmol·L⁻¹)处理更有利于生物碱的积累。腐胺含量与腐胺合成关键酶活性变化基本一致,二者均随MeJA浓度增加先升高后降低,且在200μmol·L⁻¹ MeJA时腐胺含量达到最高。进一步检测TAs合成途径中PMT、TRⅠ和H6H的基因表达,发现PMT基因表达水平无明显变化趋势。200μmol·L⁻¹ MeJA处理下,叶片和根中TRⅠ和H6H的表达水平最高。由此推测,MeJA对莨菪碱和东莨菪碱形成的调节主要是通过影响关键酶基因的表达。适当浓度的MeJA增加了叶片和根中TRⅠ以及根中H6H的基因表达,促进了生物碱的积累以及莨菪碱向东莨菪碱的转化。