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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2016, Vol. 36 Issue (9): 11-20    DOI: 10.13523/j.cb.20160902
研究报告     
中间锦鸡儿CiCHIL克隆及其黄酮代谢功能研究
韩晗, 包旦奇, 杨飞芸, 刘坤, 杨天瑞, 杨杞, 李国婧, 王瑞刚
内蒙古农业大学生命科学学院 呼和浩特 010018
The Cloning and Function Analysis in Flavonoid Pathway of CiCHIL from Caragana intermedia
HAN Han, BAO Dan-qi, YANG Fei-yun, LIU Kun, YANG Tian-rui, YANG Qi, LI Guo-jing, WANG Rui-gang
College of Life Science, Inner Mongolia Agricultual University, Hohhot 010018, China
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摘要:

植物的次生代谢产物能够为植物的果实和花着色、有助于种子的形成、花粉的传播、适应外界逆境及防御害虫的攻击。苯丙烷代谢途径是植物中一个重要的次生代谢途径,苯丙烷代谢通路上有几条主要的分支,分支下游产生了上千种化合物。查耳酮异构酶(chalcone isomerase,CHI)是苯丙烷通路上的一个关键酶,催化查耳酮到黄烷酮的反应,主要作用是帮助底物进行正确的分子内环化反应,生成的黄烷酮类化合物成为苯丙烷代谢途径下游产物的底物。从中间锦鸡儿干旱胁迫抑制性削减杂交文库中克隆得到一个CHI基因家族成员,序列分析和系统进化分析表明,该基因属于CHIL基因亚家族成员,命名为CiCHIL。实时荧光定量PCR检测发现,该基因受UV-B诱导且过表达CiCHIL具有较强的抵御紫外胁迫的能力。对过表达株系进行RNA和蛋白质水平的检测,对挑选出来的过表达株系进行总黄酮含量检测,发现过表达株系总黄酮含量显著高于野生型。

关键词: 查耳酮异构酶Western blot总黄酮含量紫外胁迫中间锦鸡儿    
Abstract:

Secondary metabolites of plants colour fruits and flowers, favouring seed and pollen dispersal, and contribute to plant adaptation to environmental conditions and pathogen attacks. The general phenylpropanoid pathway is an important secondary metabolic pathway in plant,it provides precursors for several branches leading to the elaboration of thousands of compounds. A key branch-point enzyme of the phenylpropanoid pathway, chalcone isomerase (CHI), catalyzes the reaction producing flavanones, the skeletal backbone for many downstream metabolites. This reaction is conducted by the cyclization of naringenin chalcone to naringenin. An CHI encoding gene fragment was isolated from the suppression subtractive hybridization library of Caragana intermedi Kom., sequence and phylogenetic analysis indicatead that the protein belongs to the CHIL subfamily,therefore it was desingnated as CiCHIL.Real-time quantitive PCR analysis showed that the transcript of CiCHIL was induced under UV-B treatment,and overexpression of CiCHIL enhanced the resistance to UV-B. RNA and protein expression levels were detected in the overexpression lines,determination of total flavonoids in selected overexpression lines.The results showed that the content of total flavonoids was significantly higher than that of wild type.

Key words: Western blot    CHI    Total flavonoids content    UV-B stress    Caragana intermedia
收稿日期: 2016-05-03 出版日期: 2016-09-25
ZTFLH:  Q946.83  
基金资助:

国家自然科学基金(31360169),内蒙古产业创新创业人才团队、内蒙古自治区科技创新团队(201503004)资助项目

通讯作者: 王瑞刚     E-mail: ruigangwang@126.com
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引用本文:

韩晗, 包旦奇, 杨飞芸, 刘坤, 杨天瑞, 杨杞, 李国婧, 王瑞刚. 中间锦鸡儿CiCHIL克隆及其黄酮代谢功能研究[J]. 中国生物工程杂志, 2016, 36(9): 11-20.

HAN Han, BAO Dan-qi, YANG Fei-yun, LIU Kun, YANG Tian-rui, YANG Qi, LI Guo-jing, WANG Rui-gang. The Cloning and Function Analysis in Flavonoid Pathway of CiCHIL from Caragana intermedia. China Biotechnology, 2016, 36(9): 11-20.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160902        https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I9/11

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