茶树CsCIGR基因克隆及表达特性分析
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引用本文:王 爽,王永鑫,王 瑜,李 辉,滕瑞敏,庄 静.茶树CsCIGR基因克隆及表达特性分析[J].西北植物学报,2019,39(5):867~875
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作者单位
王 爽,王永鑫,王 瑜,李 辉,滕瑞敏,庄 静* (南京农业大学 园艺学院茶叶科学研究所南京210095) 
基金项目:国家自然科学基金(31570691,31870681)
中文摘要:该研究以茶树基因组数据库为基础,采用RT PCR技术,从茶树‘龙井43’中克隆得到基因CsCIGR。序列分析显示,CsCIGR基因开放阅读框长度为1 677 bp,编码588个氨基酸。进化分析表明,CsCIGR属于GRAS家族的PAT1亚家族。多序列比对显示,茶树CsCIGR蛋白与其他植物的GRAS蛋白氨基酸序列具有很高的相似性。氨基酸理化性质分析显示,CsCIGR转录因子属于亲水性蛋白。亚细胞定位预测显示,CsCIGR可能位于细胞核中。启动子预测分析发现,CsCIGR启动子区域包含胁迫响应元件(STRE)、干旱应答元件(MYC)、厌氧诱导元件(ARE)等多种与逆境响应相关的顺式作用元件。荧光定量PCR分析结果显示,CsCIGR基因在低温(4 ℃)、高温(38 ℃)、干旱(200 g·L-1 PEG)、高盐(200 mmol·L-1 NaCl)胁迫下均能诱导表达,且对高盐,低温和高温胁迫响应更为明显,推测CsCIGR基因在茶树响应逆境胁迫中发挥重要作用。该研究为茶树抗性育种筛选基因提供了重要理论依据。
中文关键词:茶树  CsCIGR  进化分析  非生物胁迫  表达分析
 
Cloning and Expression Profiles Analysis of CsCIGR Gene in Camellia sinensis
Abstract:In this study, the CsCIGR gene was cloned from Camellia sinensis cultivar ‘Longjing 43’ by RT PCR method based on tea genome database. Sequence analysis showed that the open reading frame length of CsCIGR gene was 1 677 bp, encoding 588 amino acids. Evolutionary analysis indicated that CsCIGR belongs to the PAT1 subfamily of the GRAS family. Multiple sequence alignments indicate that the CsCIGR protein of tea plants has a high degree of similarity to the amino acid sequences of GRAS proteins from other plants. The physicochemical properties of amino acids showed that CsCIGR transcription factor was a hydrophilic proteins. Subcellular localization prediction indicates that CsCIGR may be located in the nucleus. Promoter prediction analysis showed that the promoter region of CsCIGR contained stress response element (STRE), drought response element (MYC), anaerobic inducer (ARE) and other cis acting elements related to stress response. The results of qRT PCR showed that the CsCIGR gene responded to high temperature (38 ℃), low temperature (4 ℃), drought (200 g·L-1 PEG) and high salinity(200 mmol·L-1 NaCl). The expression levels of CsCIGR were significantly induced by high salinity, low temperature, and high temperature stresses. It is speculated that CsCIGR gene plays an important role in the response of C. sinensis to stresses. The results of this study provide important theoretical basis for screening genes of C. sinensis resistance breeding.
keywords:Camellia sinensis  CsCIGR  phylogenetic analysis  abiotic stress  expression analysis
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