地黄次生代谢产物生物合成基因表达水平与其含量的相关性分析

doi:10.13304/j.nykjdb.2017.0664

中国农业科技导报 ›› 2018, Vol. 20 ›› Issue (11): 36-43.DOI: 10.13304/j.nykjdb.2017.0664

地黄次生代谢产物生物合成基因表达水平与其含量的相关性分析

朱畇昊1,2,李璐1,赵乐1,2,董诚明1,2*

1.河南中医药大学药学院, 郑州 450046; 2.呼吸疾病诊疗与新药研发河南省协同创新中心, 郑州 450046

收稿日期:2017-09-29 出版日期:2018-11-15 发布日期:2018-04-28
通讯作者: *通信作者：董诚明，教授，主要从事中药规范化种植研究。E-mail：dcm371@sohu.com
作者简介:朱畇昊，讲师，博士，主要从事药用植物分子生物学研究。E-mail：guxinhan123@163.com。
基金资助:
国家自然科学基金项目（81603232）；国家重点研发计划项目（2017YFC1702800）；河南省科技攻关项目（172102310539）；河南省重大科技专项（171100310500）；河南中医学院博士科研基金项目（BSJJ2015-13）资助。

Correlation Analysis Between Expression Levels of Biosynthetic Genes for Secondary Metabolites and Their Contents in Rehmannia glutinosa

ZHU Yunhao1,2, LI Lu1, ZHAO Le1,2, DONG Chengming1,2*

1.College of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046; 2.Collaborative Innovation Center for Respiratory Disease Diagnosis and Treatment & Chinese Medicine Development of Henan Province, Zhengzhou 450046, China

Received:2017-09-29 Online:2018-11-15 Published:2018-04-28

摘要/Abstract

摘要： 为了研究地黄次生代谢产物生物合成基因在不同地黄材料中的相对表达量，并分析其与次生代谢产物积累的关系，从地黄叶、地黄组培苗叶、地黄毛状根中提取总RNA，以地黄TIP41为内参基因，利用实时荧光定量PCR技术分析梓醇、毛蕊花糖苷生物合成相关酶基因的表达量，并分别采用高效液相色谱法测定3种材料中梓醇和毛蕊花糖苷的含量、紫外分光光度法测定总环烯醚萜苷的含量，再对两者进行相关性分析。结果表明，3种不同地黄材料中次生代谢产物的生物合成相关酶基因的表达量及其含量差异较大。经相关性分析，地黄中梓醇的含量与GGPPS2基因的表达量呈显著正相关，总环烯醚萜苷的含量与GGPPS1、G10H基因的表达量也有显著相关性，说明GGPPS2与梓醇的合成有关，GGPPS1、G10H与总环烯醚萜苷的合成有关；而毛蕊花糖苷的含量与ADT、ASP5、4CL、C3H、HCT、GOT2基因的表达量均没有显著相关性。上述结果为地黄次生代谢产物生物合成途径的相关研究提供了参考。

关键词: 地黄, 次生代谢产物, 生物合成, 基因表达, 实时荧光定量PCR

Abstract: In order to study the relative expression of the biosynthetic genes for secondary metabolites in different materials of Rehmannia glutinosa, and analyze the relationship between gene expression and secondary metabolites accumulation, this paper extracted total RNA from leaves, leaves of tissue culture seeding and hairy roots of R. glutinosa, respectively. TIP41 was used as a reference gene, and the relative expression of the biosynthesis-related enzymes genes for catalpol and verbascoside were analyzed by real-time quantitative PCR. The content of catalpol and verbascoside in 3 kinds of material were measured by HPLC, respectively, while the content of total iridoid glycoside was measured by UV spectrophotometry, and then the correlation between their contents and relative expressions of the genes related to biosynthesis enzymes were analyzed. The results indicated that there were more differences in the expression levels of the genes related to biosynthesis enzyme for secondary metabolites and their contents among the 3 different kinds of R. glutinosa materials. According to relative correlation analysis, the content of catalpol was significantly and positively correlated with the expression quantity of GGPPS2, and the content of total iridoid glycoside was also significantly correlated with the expression quantities of GGPPS1 and G10H, indicating that GGPPS2 was related with the synthesis of catalpa, GGPPS1 and G10H might be related to total iridoid glycoside biosynthesis. While, there were no significant correlation between the content of verbascoside with the expression levels of ADT, ASP5, 4CL, C3H, HCT and GOT2. These results provided references for relative studies on secondary metabolites biosynthetic pathways of R. glutinosa.

朱畇昊1,2,李璐1,赵乐1,2,董诚明1,2*. 地黄次生代谢产物生物合成基因表达水平与其含量的相关性分析[J]. 中国农业科技导报, 2018, 20(11): 36-43.

ZHU Yunhao1,2, LI Lu1, ZHAO Le1,2, DONG Chengming1,2*. Correlation Analysis Between Expression Levels of Biosynthetic Genes for Secondary Metabolites and Their Contents in Rehmannia glutinosa[J]. Journal of Agricultural Science and Technology, 2018, 20(11): 36-43.

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地黄次生代谢产物生物合成基因表达水平与其含量的相关性分析

Correlation Analysis Between Expression Levels of Biosynthetic Genes for Secondary Metabolites and Their Contents in Rehmannia glutinosa

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