茶树CsGIF1基因的克隆及其对非生物胁迫的响应分析
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国家自然科学基金(31570691)


Cloning and Expression Analysis of CsGIF1 Gene under Abiotic Stress in Camellia sinensis
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    摘要:

    GRFINTERACTING FACTOR (GIF)基因是植物叶发育相关的一类重要调控因子,调节植物叶器官的发育。该研究采用RTPCR方法从茶树‘龙井43’的叶片cDNA中克隆得到CsGIF1基因,并利用荧光定量PCR分析了高温(38 ℃)、低温(4 ℃)、干旱(200 g·L-1 PEG)、盐胁迫(200 mmol·L-1 NaCl)下CsGIF1基因的表达水平,以明确CsGIF1基因对非生物胁迫的应答特性,为茶树CsGIF1基因的逆境调控以及功能研究奠定基础。结果表明:(1)CsGIF1基因长666 bp,编码221个氨基酸,具有高度保守的SNH结构域;CsGIF1蛋白为亲水性蛋白,理论相对分子质量为23 380,理论等电点为6.30;酸性氨基酸、碱性氨基酸、芳香族氨基酸和脂肪族氨基酸所占比例分别为7%、9%、5%和13%;蛋白质二级结构显示,茶树CsGIF1蛋白由38.01%的α螺旋、10.41%的β折叠、12.22%的延伸主链和39.37%的随机卷曲组成。(2)qRTPCR分析显示,茶树CsGIF1基因对于4种非生物胁迫均有响应,但不同处理响应不同。其中在胁迫2 h时,高温和低温胁迫下,CsGIF1基因相对表达量显著增加,分别为对照的3.54和5.69倍,且高低温胁迫下,CsGIF1基因响应明显大于干旱和盐胁迫。在高温、低温、干旱、盐等不同胁迫处理下,茶树‘龙井43’中CsGIF1基因的表达水平与对照相比均差异明显。

    Abstract:

    GRFINTERACTING FACTOR (GIF) gene played important roles in regulating the development of leaf. GIF gene could regulate cell proliferation and expansion to form different leaf sizes. In this study, the gene of CsGIF1 was cloned using RTPCR from leaf cDNA template of tea plant cultivar ‘Longjing43’. In order to clarify the response characteristics of CsGIF1 to abiotic stress, we performed qRTPCR to analyze the expression levels of CsGIF1 under the stresses of high temperature (38 ℃), low temperature (4 ℃), drought (200 g·L-1 PEG) and salt (200 mmol·L-1). This study could lay the foundation for the stress regulation and function study of CsGIF1 in tea plant. The results indicated that: (1) the length of CsGIF1 was 666 bp, encoding 221 amino acids with a highly conserved region called SNH domain. Protein analysis suggested that the CsGIF1 was hydrophilic protein, and its theoretical relative molecular weight is 23 380, theoretical isoelectric point is 6.30, the percentages of acidic, basic, aromatic and aliphatic amino acids are 7%, 9%, 5% and 13%, respectively. Secondary structure prediction showed that CsGIF1 protein consists of 38.01% alphahelix, 10.41% beta turn, 12.22% extended strand and 39.37% random coil, respectively. (2) qRTPCR analysis demonstrated that CsGIF1 gene could respond to the all four abiotic stresses. However, the CsGIF1 gene presented different expression trends to those abiotic stresses. At 2h after the treatment of high or low temperature, the relative expression levels of CsGIF1 gene were upregulated significantly, which was 3.54 and 5.69 times of the control, respectively. Under high or low temperature stress, the response of CsGIF1 gene was significantly greater than under drought or salt stress. Under the treatments of high temperature, low temperature, drought and salt, the expression levels of CsGIF1 showed significant difference in the leaves of C. sinensis cv. ‘Longjing43’.

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王雅慧,王永鑫,王文丽,等.茶树CsGIF1基因的克隆及其对非生物胁迫的响应分析[J].西北植物学报,2017,37(12):2363-2369

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  • 在线发布日期: 2017-12-29
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