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太子参玉米黄质环氧化酶基因全长CDS的克隆与表达分析

[Cloning and expression analysis on full length CDS of zeaxanthin epoxidase gene in Pseudostellaria heterophylla].

作者信息

Zheng Wei, Zhou Tao, Li Jun, Long Deng-Kai, Jiang Wei-Ke, Ding Ling

机构信息

Guiyang University of Chinese Medicine, Guiyang 550002, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2017 Feb;42(4):669-674. doi: 10.19540/j.cnki.cjcmm.20170103.034.

Abstract

Zeaxanthin epoxidase plays an important role in indirect pathway of plant abscisic acid biosynthesis. According to the data of Pseudostellaria heterophylla transcriptome, zeaxanthin epoxidase gene was isolated and named as PhZEP. The results of bioinformatics analysis showed that the coding sequence of PhZEP was 1 263 bp long and encoded 420 amino acids. The putative protein molecular weight was 47.34 kDa and its theoretical isoelectric point was 6.64. The characteristic structure domains were predicted, including binding site of lipoprotein and flavoprotein monooxyenase. A signal peptide was discovered at the N-terminal of amino acids. The Real-time PCR revealed that PhZEP had a higher expression level in leaves than other tissues of P.heterophylla. Highly expressed PhZEP was also observed at 10 d and 40 d tuberous root after flowering. PhZEP presented a different expression after treatment with ABA, fluridone and ABA +fluridone compared to the control. The expression of PhZEP in tuberous root after ABA treatment was close to that in control group, while PhZEP showed significant up-regulation in the fluridone treatment group. In this study, the PhZEP gene from P. heterophylla was cloned and this result has important significance for its functional identification. This research provides a basis for the further analysis on functional mechanism of ABA during development of P. heterophylla.

摘要

玉米黄质环氧化酶在植物脱落酸生物合成的间接途径中起重要作用。根据太子参转录组数据,分离出玉米黄质环氧化酶基因并命名为PhZEP。生物信息学分析结果表明,PhZEP的编码序列长1263 bp,编码420个氨基酸。推测的蛋白质分子量为47.34 kDa,理论等电点为6.64。预测了其特征结构域,包括脂蛋白结合位点和黄素蛋白单加氧酶。在氨基酸的N端发现了一个信号肽。实时荧光定量PCR显示,PhZEP在太子参叶片中的表达水平高于其他组织。在开花后的10 d和40 d块根中也观察到PhZEP的高表达。与对照相比,ABA、氟啶酮和ABA +氟啶酮处理后PhZEP呈现不同的表达。ABA处理后块根中PhZEP的表达与对照组接近,而氟啶酮处理组中PhZEP显著上调。本研究克隆了太子参的PhZEP基因,该结果对其功能鉴定具有重要意义。本研究为进一步分析脱落酸在太子参发育过程中的作用机制提供了依据。

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