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[黄芪皂苷生物合成过程对短期水分变化的响应]

[Short-term water changes response of saponin biosynthesis process in Astragalus membranceus].

作者信息

Wei He, Cheng Lin, Wu Pei, Han Mei, Yang Li-Min

机构信息

College of Chinese Materials, Jilin Agricultural University Changchun 130118, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2019 Feb;44(3):441-447. doi: 10.19540/j.cnki.cjcmm.20181204.008.

Abstract

The study is aimed to explore the effect of different water on the content of total saponins,astragaloside Ⅳ and gene expression in the growth of Astragalus membranceus. In this study, one-year-old A. membranaceus was used as the experimental material, by pot culture different water treatments were simulated at herbal garden in Jilin Agricultural University. The content of astragaloside Ⅳ was determined by HPLC and the total saponins by UV spectrophotometry. With 18 S RNA as a reference gene, fluorescent quantitative PCR was applied to analyze the eight key enzymes in astragalus saponin synthesis pathway AACT,HMGS,HMGR,IDI,FPS,SS,SE,CAS expression. With the decrease of soil water, the content of astragaloside Ⅳ in the root tissue of A. membranaceus showed an increasing trend, up to 1.46 mg·g(-1). The total saponin content tended to increase, up to 6.80 mg·g(-1). The results of relative expression of genes showed that the eight genes showed different effects at different water. With the change of soil water content, the amount of(AACT,IDI,SS) relative expression in drought stress group firstly increased and then decreased, then increased, and then decreased. The amount of(HMGS,HMGR,FPS) relative expression in drought stress group increased firstly and then decreased. The amount of(SE,CAS) relative expression in drought stress group increased firstly and then decreased, and continued to decrease after rehydration. The expression of key enzyme genes involved in the synthesis of astragaloside was influenced by each other, and the expression of key enzyme in roots showed a correlation with the content of astragaloside. Correlation analysis showed that there was a very significant positive correlation between HMGR gene and total saponins content in drought stress group and a significant negative correlation between content of CAS and total saponins. The contents of FPS,SE,CAS and astragaloside Ⅳ were very significantly and negative correlated. The relationship between other genes and quality was positive. Therefore, HMGR, FPS, SE and CAS genes have significant effects on the regulation of saponin content under water control. On the 15 th day after water regulation, the total amount of astragaloside and total saponins reached the highest value and could be harvested.

摘要

本研究旨在探讨不同水分条件对黄芪生长过程中总皂苷、黄芪甲苷含量及基因表达的影响。本研究以一年生黄芪为实验材料,在吉林农业大学药园通过盆栽模拟不同水分处理。采用高效液相色谱法测定黄芪甲苷含量,紫外分光光度法测定总皂苷含量。以18S RNA为内参基因,运用荧光定量PCR分析黄芪皂苷合成途径中8个关键酶AACT、HMGS、HMGR、IDI、FPS、SS、SE、CAS的表达。随着土壤水分的降低,黄芪根组织中黄芪甲苷含量呈上升趋势,最高可达1.46mg·g(-1)。总皂苷含量也呈上升趋势,最高可达6.80mg·g(-1)。基因相对表达结果表明,8个基因在不同水分条件下表现出不同的效应。随着土壤含水量的变化,干旱胁迫组中(AACT、IDI、SS)的相对表达量先升高后降低,再升高,再降低。干旱胁迫组中(HMGS、HMGR、FPS)的相对表达量先升高后降低。干旱胁迫组中(SE、CAS)的相对表达量先升高后降低,复水后继续降低。黄芪甲苷合成相关关键酶基因的表达相互影响,根中关键酶的表达与黄芪甲苷含量存在相关性。相关性分析表明,干旱胁迫组中HMGR基因与总皂苷含量呈极显著正相关,CAS含量与总皂苷呈显著负相关。FPS、SE、CAS与黄芪甲苷含量呈极显著负相关。其他基因与品质的关系为正相关。因此,HMGR、FPS、SE和CAS基因在水分调控下对皂苷含量的调节具有显著作用。水分调控后第15天,黄芪甲苷和总皂苷总量达到最高值,可以采收。

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