《植物生理学报》 2017, 53(1): 38-44

茶树发育相关基因CsARGOS的克隆与表达特性分析

王文丽, 吴致君, 刘志薇, 崔新, 庄静^*

南京农业大学园艺学院, 茶叶科学研究所, 南京210095

通信作者：庄；E-mail: zhuangjing@njau.edu.cn

摘　要：

植物发育相关基因ARGOS基因受生长素诱导, 在植物的生长发育和激素调控方面起着重要的作用。本实验选取茶树品种‘龙井43’作为试验材料, 利用RT-PCR方法, 从茶树cDNA中克隆得到茶树ARGOS基因, 命名为CsARGOS。序列分析表明, CsARGOS基因开放阅读框为366 bp, 编码121个氨基酸。进化树分析显示, CsARGOS蛋白与双子叶植物的进化树关系相对较近, 与单子叶植物的进化树关系相对较远。序列多重比对显示, CsARGOS蛋白具有ARGOS蛋白的特征基序, C端均含有丰富的亮氨酸, N端亲水性氨基酸居多, 这些区域在物种中是相对保守的。氨基酸组分、理化性质、亲水性/疏水性、二级结构预测和无序化分析显示, 茶树CsARGOS是疏水性蛋白, 以随机卷曲为主, 无序化特征不明显。利用实时荧光定量PCR对CsARGOS基因在茶树中的表达进行检测, 分析结果表明在不同组织中, CsARGOS基因的表达水平从高到低依次为根>第三叶>第四叶>老叶>嫩茎>第二叶>第一叶; 不同激素处理下, CsARGOS的表达水平从高到低依次为GA3>SA>ABA>MeJA>IAA。

关键词：茶树; ARGOS; 进化树分析; 不同组织; 激素处理

收稿：2016-09-02   修定：2016-10-24

资助：国家自然科学基金(31570691)。

Cloning and expression profiles analysis of development related gene CsARGOS in tea plant (Camellia sinensis)

WANG Wen-Li, WU Zhi-Jun, LIU Zhi-Wei, CUI Xin, ZHUANG Jing^*

Tea Science Research Institute, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China

Corresponding author: ZHUANG Jing; E-mail: zhuangjing@njau.edu.cn

Abstract:

ARGOS gene is induced by auxin and plays important roles in plant development and hormonal regulation. In this study, a gene encoding ARGOS-like protein was cloned by RT-PCR method using a cDNA template from tea plant (Camellia sinensis) cultivar ‘Longjing43’. This gene is named as CsARGOS. Sequence analysis showed that the open reading frame of CsARGOS gene has 366 bp length, which encoding 121 amino acids. Analysis of phylogenetic tree showed that CsARGOS protein has high similarity with dicotyledon plants and has low similarity with the monocotyledon plant. Multiple alignments showed that CsARGOS protein contains Leu-rich domain in the C-terminal and rich hydrophilic amino acids in N-terminal, which are the characterized motifs of ARGOS-type protein. Amino acid composition, physical and chemical properties, hydrophilicity/hydrophobicity, secondary structure predicted and disordered residues of CsARGOS protein were analyzed. The disordered residues of CsARGOS protein are not obvious. Most amino acids of CsARGOS protein are hydrophobic. The secondary structure of CsARGOS is mainly random coil. The expression levels of CsARGOS gene in tea plant were detected using quantitative real-time PCR analysis. In different tissues, the successive order of CsARGOS gene expression levels was root, the third leaf, the fourth leaf, old leaf, stem, the second leaf, and the first leaf. In different hormonal treatments, the successive order of CsARGOS gene expression levels was GA3, SA, ABA, MeJA, and IAA.

Key words: Camellia sinensis; ARGOS; phylogenetic tree analysis; different tissues; hormonal treatment