茶树CsGME1基因的克隆与逆境胁迫响应分析
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国家自然科学基金(31870681);


Cloning and Response Analysis to Abiotic Stress of CsGME1 Gene in Camellia sinensis
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    摘要:

    该研究基于茶树转录组和基因组信息,以茶树‘龙井43’为实验材料,从其cDNA中克隆获得茶树CsGME1基因,并对其蛋白序列特征、基因表达模式及其在不同非生物胁迫下的表达水平进行实时荧光定量分析。结果显示:(1)茶树CsGME1开放阅读框长度为1 131 bp,编码376个氨基酸;该序列与多个相关物种的GME氨基酸序列一致性为94.25%,均含有NAD结合域。(2)进化树分析表明,茶树CsGME1基因与番茄SlGME1亲缘关系较近,与水稻OsGME2亲缘关系最远。(3)CsGME1蛋白属于亲水性蛋白,理论相对分子量为42 046.84 Da,理论等电点为5.73,具有4个无序化区域,无序化程度较低;CsGME1蛋白二级结构由39.25% α螺旋,13.26%延伸主链,5.84% β转角和41.38%随机卷曲组成;三级结构分析结果显示,CsGME1包含螺旋和随机卷曲,与二级结构吻合。(4)荧光定量分析结果显示,在高温(38 ℃)、低温(4 ℃)、干旱(20% PEG)和高盐(200 mmol·L-1 NaCl)4种非生物胁迫处理下,茶树CsGME1基因均有响应,且表达存在差异,推测CsGME1参与了茶树的逆境胁迫响应过程。

    Abstract:

    In this study, the CsGME1 gene was cloned from cDNA of Camellia sinensis cv. ‘Longjing43’ based on the transcriptome and genome database of tea plant. Its protein characteristics and gene expression patterns were analyzed. In addition, the expression profiles of the CsGME1 under abiotic stress treatments in tea plant were detected by quantitative real time PCR. The main results listed below: (1) sequence analysis showed that the length of CsGME1 open reading frame (ORF) of tea plant was 1131 bp, encoding 376 amino acids. Multiple alignments of CsGME1 with related species showed that the identity of them was 94.25%, and all contain NAD binding domain. (2) Analysis of phylogenetic tree indicated that the CsGME1 was closely related to SlGME1 from Solanum lycopersicum, but far from OsGME2 of Oryza sativa. (3) Protein analysis suggested that the CsGME1 is a hydrophilic protein with the theoretical relative molecular weight 42 046.84 Da and the theoretical isoelectric point 5.73. CsGME1 protein had 4 disordered regions with low degree of disorder. Secondary structure prediction showed that the CsGME1 protein consists of 39.25% αhelix, 13.26% extended strand, 5.84% βturn and 41.38% random coil. The threedimensional structures indicated that CsGME1 protein contains helix and random coil. (4) The results of quantitative real time PCR showed that the CsGME1 gene responded to four kinds of abiotic stress treatments: high temperature (38 ℃), low temperature (4 ℃), drought (20% PEG) and high salinity (200 mmol·m-1 NaCl). The expression profiles showed difference.

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杨雅焯,李 辉,林士佳,等.茶树CsGME1基因的克隆与逆境胁迫响应分析[J].西北植物学报,2020,40(2):232-239

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  • 在线发布日期: 2020-05-02
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