雨生红球藻转录因子Whirly1的获得及其表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2020-1348

摘要/Abstract

摘要：

Whirly转录因子广泛存在于植物中,参与植物生长发育进程和逆境胁迫耐受,对其功能的研究有助于我们理解雨生红球藻响应胁迫的机制。因此,先根据雨生红球藻192.80转录组测序数据,分离克隆到一个Whirly转录因子序列,并对其结构及表达进行分析。该转录因子的cDNA序列为924 bp,基因组序列为2 556 bp,由有5个外显子和4个内含子组成,命名为HpWhirly1。HpWhirly1预测由244个氨基酸组成,拥有Whirly转录因子的保守结构域。HpWhirly1在醋酸钠和水杨酸胁迫早期表达上调,在强光-醋酸钠联合胁迫和缺氮胁迫条件下表达下调,表明其在雨生红球藻受到胁迫时发挥一定的作用。以上研究结果将有助于进一步研究Whirly转录因子的功能及其在雨生红球藻响应胁迫时的作用。

关键词: 雨生红球藻, Whirly, 转录因子, 克隆, 表达分析

Abstract:

Whirly transcription factors are found in plants ubiquitously,and play a role in plant growth and stress resistance. Uncovering the functions of Whirly transcription factors would help us understand the response mechanism to stress in Haematococcus pluvialis. Hence,a Whirly transcription factor named HpWhirly1 was isolated and cloned based on the transcriptome data of H. pluvialis 192.80 and its structure and expression were analyzed. The cDNA sequence of this transcription factor consisted of 924 bp and genome sequence was 2 556 bp with 5 exons and 4 introns. Results showed that HpWhirly1 was predicted to contain 244 amino acids and had the conserved domain of Whirly transcription factor. Further analysis showed that the expression of HpWhirly1 was up-regulated during the early stage of sodium acetate and salicylic acid stresses,while it was down-regulated under strong light sodium acetate combined stress and nitrogen deficiency stress. The above results indicated that HpWhirly1 might play a role in H. pluvialis when facing stresses and could respond to them. The results of this study are conducive to further studying the functions of Whirly transcription factors and its effects on the stress response of H. pluvialis.

Key words: Haematococcus pluvialis, Whirly, transcription factor, molecular cloning, transcription profile

刘纹甫, 黄丹琼, 胡群菊, 王潮岗. 雨生红球藻转录因子Whirly1的获得及其表达分析[J]. 生物技术通报, 2021, 37(11): 248-256.

LIU Wen-fu, HUANG Dan-qiong, HU Qun-ju, WANG Chao-gang. Acquisition and Expression Analysis of the Transcription Factor Whirly1 from Haematococcus pluvialis[J]. Biotechnology Bulletin, 2021, 37(11): 248-256.

图/表 8

图1 雨生红球藻HpWhirly1基因的克隆

Fig. 1 Cloning of HpWhirly1 gene from Haematococcus pluvialis

图2 HpWhirly1的氨基酸序列

Fig. 2 Amino acid sequence of HpWhirly1

图3 HpWhirly1基因结构分析橙色部分为外显子,蓝色部分为UTR区域,黑线为内含子

Fig.3 Gene structure analysis of HpWhirly1 The orange parts are exons,the blue parts are UTR regions,and the black lines are introns

图4 HpWhirly1保守结构域分析

Fig.4 Conserved domain analysis of HpWhirly1

图5 HpWhirly1蛋白三级结构预测

Fig.5 Predicted 3D structure of HpWhirly1

图6 HpWhirly1与其他物种的Whirly同源性分析 Haematococcus lacustris（GFH09517.1）,Chlamydomonas eustigma（GAX74163.1）,Dunaliella salina（KAF5835724.1）,Micractinium conductrix（PSC72124.1）;以及模式生物拟南芥中3个Whirly转录因子序列:AtWHY1（AT1G14410）,AtWHY2（AT1G71260）,AtWHY3（AT2G02740）

Fig.6 Homology analysis of HpWhirly1 and other WHYs from different species Haematococcus lacustris（GFH09517.1）,Chlamydomonas eustigma（GAX74163.1）,Dunaliella salina（KAF5835724.1）,and Micractinium conductrix（PSC72124.1）. Three Whirly transcription factor sequences in the model organism Arabidopsis:AtWHY1（AT1G14410）,AtWHY2（AT1G71260）,and AtWHY3（AT2G02740）

图7 Whirly转录因子的系统进化树分析

Fig.7 Phylogenetic tree analysis of the Whirly transcription factor

图8 不同胁迫条件下HpWhirly1的表达分析 A:强光处理;B:醋酸处理;C:强光醋酸钠联合处理;D:水杨酸处理;E:缺氮处理。“*”表示处理组和对照组的表达水平存在显著性差异（P<0.05）;“**”表示存在极显著性差异（P<0.01）

Fig.8 Expression analysis of HpWhirly1 under different stress conditions A: Strong light treatment. B: Acetic acid treatment. C: Strong light sodium acetate combined treatment. D: Salicylic acid treatment. E: Nitrogen deficiency treatment. * indicates significant difference between treatment group and control group（P<0.05）. ** indicates extremely significant difference between treatment group and control group（P<0.01）

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