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CLC number: Q943.2

On-line Access: 2017-08-08

Received: 2016-05-17

Revision Accepted: 2016-09-10

Crosschecked: 2017-07-25

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Jing-ya Zhao


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Journal of Zhejiang University SCIENCE B 2017 Vol.18 No.8 P.662-673


Molecular cloning, characterization, and promoter analysis of the isochorismate synthase (AaICS1) gene from Artemisia annua

Author(s):  Lu-yao Wang, Ying Zhang, Xue-qing Fu, Ting-ting Zhang, Jia-wei Ma, Li-da Zhang, Hong-mei Qian, Ke-xuan Tang, Shan Li, Jing-ya Zhao

Affiliation(s):  Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; more

Corresponding email(s):   zhaojy@sjtu.edu.cn

Key Words:  Salicylic acid, Artemisia annua L., Quantitative real-time polymerase chain reaction (qRT-PCR), Isochorismate synthase

Lu-yao Wang, Ying Zhang, Xue-qing Fu, Ting-ting Zhang, Jia-wei Ma, Li-da Zhang, Hong-mei Qian, Ke-xuan Tang, Shan Li, Jing-ya Zhao. Molecular cloning, characterization, and promoter analysis of the isochorismate synthase (AaICS1) gene from Artemisia annua[J]. Journal of Zhejiang University Science B, 2017, 18(8): 662-673.

@article{title="Molecular cloning, characterization, and promoter analysis of the isochorismate synthase (AaICS1) gene from Artemisia annua",
author="Lu-yao Wang, Ying Zhang, Xue-qing Fu, Ting-ting Zhang, Jia-wei Ma, Li-da Zhang, Hong-mei Qian, Ke-xuan Tang, Shan Li, Jing-ya Zhao",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Molecular cloning, characterization, and promoter analysis of the isochorismate synthase (AaICS1) gene from Artemisia annua
%A Lu-yao Wang
%A Ying Zhang
%A Xue-qing Fu
%A Ting-ting Zhang
%A Jia-wei Ma
%A Li-da Zhang
%A Hong-mei Qian
%A Ke-xuan Tang
%A Shan Li
%A Jing-ya Zhao
%J Journal of Zhejiang University SCIENCE B
%V 18
%N 8
%P 662-673
%@ 1673-1581
%D 2017
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1600223

T1 - Molecular cloning, characterization, and promoter analysis of the isochorismate synthase (AaICS1) gene from Artemisia annua
A1 - Lu-yao Wang
A1 - Ying Zhang
A1 - Xue-qing Fu
A1 - Ting-ting Zhang
A1 - Jia-wei Ma
A1 - Li-da Zhang
A1 - Hong-mei Qian
A1 - Ke-xuan Tang
A1 - Shan Li
A1 - Jing-ya Zhao
J0 - Journal of Zhejiang University Science B
VL - 18
IS - 8
SP - 662
EP - 673
%@ 1673-1581
Y1 - 2017
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1600223

isochorismate synthase (ICS) is a crucial enzyme in the salicylic acid (SA) synthesis pathway. The full-length complementary DNA (cDNA) sequence of the ICS gene was isolated from Artemisia annua L. The gene, named AaICS1, contained a 1710-bp open reading frame, which encoded a protein with 570 amino acids. Bioinformatics and comparative study revealed that the polypeptide protein of AaICS1 had high homology with ICSs from other plant species. Southern blot analysis suggested that AaICS1 might be a single-copy gene. Analysis of the 1470-bp promoter of AaICS1 identified distinct cis-acting regulatory elements, including TC-rich repeats, MYB binding site (MBS), and TCA-elements. An analysis of AaICS1 transcript levels in multifarious tissues of A. annua using quantitative real-time polymerase chain reaction (qRT-PCR) showed that old leaves had the highest transcription levels. AaICS1 was up-regulated under wounding, drought, salinity, and SA treatments. This was corroborated by the presence of the predicted cis-acting elements in the promoter region of AaICS1. Overexpressing transgenic plants and RNA interference transgenic lines of AaICS1 were generated and their expression was compared. High-performance liquid chromatography (HPLC) results from leaf tissue of transgenic A. annua showed an increase in artemisinin content in the overexpressing plants. These results confirm that AaICS1 is involved in the isochorismate pathway.


结论:AaICS1含一个总长为1710 bp的完整阅读框,编码570个氨基酸,与其它植物的ICS基因具有较高的相似性。Southern杂交结果表明AaICS1为单拷贝(图4),qRT-PCR结果显示该基因能够响应伤害、干旱、盐胁迫和水杨酸的处理,处理后基因表达量提高(图6),和启动子作用元件预测相符。qRT-PCR结果显示过表达转基因青蒿中AaICS1表达量提高,干扰转基因青蒿中该基因表达量降低(图7)。HPLC显示过表达AaICS1转基因植株中青蒿素含量提升,最高可达对照的1.9倍(图8)。


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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