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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2017, Vol. 37 Issue (12): 27-33    DOI: 10.13523/j.cb.20171206
研究报告     
异源表达CiRS基因通过生成白藜芦醇增强拟南芥的抗氧化能力 *
杨飞芸1,2,武燕燕2,崔爽1,张秀娟3,王瑞刚1,李国婧1()
1 内蒙古农业大学生命科学学院　呼和浩特　010018
2 内蒙古农业大学食品科学与工程学院　呼和浩特　010018
3 内蒙古自治区生物技术研究院　呼和浩特　010070
Heterologous Expression of CiRS Gene Eenhances the Antioxidant Capacity of Arabidopsis by Increasing the Content of Resveratrol
Fei-yun YANG1,2,Yan-yan WU2,Shuang CUI1,Xiu-juan ZHANG3,Rui-gang WANG1,Guo-jing LI1()
1 College of Life Sciences, Inner Mongolia Agricultural University, Hohhot　010018, China
2 College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot　010018, China
3 Inner Mongolia Autonomous Region Biotechnology Research Institute, Hohhot　010070, China
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摘要：

白藜芦醇是一种具有多种医疗保健作用的植物芪类次生代谢产物,在农业、医药、食品和化妆品等领域受到广泛的关注。白藜芦醇合酶是白藜芦醇生物合成中唯一必需的关键酶,决定植物体内白藜芦醇的合成。将中间锦鸡儿中克隆到的CiRS基因(GenBank登录号MF678590)转入野生型拟南芥,实验结果显示:野生型的总黄酮含量明显高于转基因株系。HPLC测得转基因拟南芥中有白藜芦醇的生成,并且含量最高达335μg/g FW。紫外照射处理后转基因植物中丙二醛的积累量明显少于野生型。转基因植物提取物DPPH自由基清除能力均高于野生型。这些结果表明,中间锦鸡儿CiRS基因异源表达后利用与黄酮类物质的共同底物合成了白藜芦醇,使得转基因植物的抗氧化性增强。
关键词： 白藜芦醇合酶;  中间锦鸡儿;  白藜芦醇;  转基因拟南芥    
Abstract:

Resveratrol is a member of stilbenoids with disease resistant activity for plants and exhibits a wide range of important biological and pharmacological properties for human which has received extensive attention in the fields of agriculture, medicine, foods, cosmetics and so on. Resveratrol synthase (RS) is an exclusive necessary enzyme in the pathway of resveratrol biosynthesis which determines the synthesis of resveratrol in plants. CiRS, a RS gene isolated from Caragana intermedia, was transferred into Arabidopsis. Total flavonoids experimental results showed that the total flavonoids content of wild type was significantly higher than that of the transgenic lines. HPLC method was used to analyze the resveratrol content in transgenic plants which maximum content was 335μg/g FW. Accumulation of malondialdehyde (MDA) after UV treatment in transgenic plants was significantly less than the wild type. DPPH free radical scavenging ability of transgenic plants extraction was higher than the wild type. Taken together, these results indicated that the antioxidant activity of transgenic plants was enhanced with the expression of CiRS gene which synthesise resveratrol with flavonoid substrates.
Key words: Resveratrol synthase    Caragana intermedia    Resveratrol    Transgenic    Arabidopsis
收稿日期: 2017-08-24 出版日期: 2017-12-16
ZTFLH:  Q786  
基金资助: 内蒙古自治区自然科学基金项目(2017MS0354);内蒙古自治区科技创新团队计划(201503004)
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杨飞芸
武燕燕
崔爽
张秀娟
王瑞刚
李国婧

引用本文:

杨飞芸,武燕燕,崔爽,张秀娟,王瑞刚,李国婧. 异源表达CiRS基因通过生成白藜芦醇增强拟南芥的抗氧化能力 *[J]. 中国生物工程杂志, 2017, 37(12): 27-33.

Fei-yun YANG,Yan-yan WU,Shuang CUI,Xiu-juan ZHANG,Rui-gang WANG,Guo-jing LI. Heterologous Expression of CiRS Gene Eenhances the Antioxidant Capacity of Arabidopsis by Increasing the Content of Resveratrol. China Biotechnology, 2017, 37(12): 27-33.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20171206    或    https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I12/27

图1  pCanG-CiRS表达载体
图2  CiRS转基因株系表达水平检测
图3  野生型和转CiRS基因拟南芥总黄酮含量比较
图4  拟南芥各株系白藜芦醇色谱图
株系名 WT OE-1 OE-16 OE-17
含量(μg/g FW) 未检出 329 313 335
表1  拟南芥各株系白藜芦醇含量
图5  拟南芥各株系MDA含量变化
图6  拟南芥各株系DPPH自由基的清除率
图7  Trolox对DPPH自由基清除率标准曲线
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