茶树叶发育相关的CsGIF1基因克隆与功能鉴定
时间:2018-12-17 17:57 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：茶树（Camellia sinensis（L.）O. Kuntze）是一种重要的叶用木本作物，可用于生产天然非酒精饮料，其鲜叶产量与经济效益密切相关。GRF-INTERACTING FACTOR (GIF)基因是叶细胞分裂过程的重要调控因子，可以与GRF基因相互作用正向调节植物叶器官的发育。本研究克隆和鉴定了茶树叶中CsGIF1基因。结果表明，茶树的CsGIF1基因全长666 bp，编码221个氨基酸，具有高度保守的SNH结构域。通过蛋白质分析可知，茶树GIF蛋白为亲水性蛋白，理论相对分子质量为23,380，理论等电点pI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%。蛋白质二级结构分析显示，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。CsGIF1基因的表达水平在嫩叶中较高，在茶叶发育的过程中持续下调。这些结果表明，CsGIF1基因可能积极参与茶树早期叶组织的形成。CsGIF1基因对不同激素刺激的反应差异揭示了其在激素调节中可能存在多重功能。本研究提供了CsGIF1基因的分子基础，可以用于未来茶树叶发育和激素刺激的功能研究。31590 毕业论文关键词：茶树；CsGIF1；叶发育；激素刺激 Cloning and functional identification of leaf development-related CsGIF1 genes in Camellia sinensis Abstract： Tea plant (Camellia sinensis (L.) O. Kuntze) is an important leaf-type woody crop used for producing of natural non-alcoholic beverages. The yield of tea plant leaf is closely related to the economic benefit. GRF-INTERACTING FACTOR (GIF) genes play an important role in regulating the process of leaf expansion. It can positively regulate the leaf development cooperating with GROWTH-INTERACTING FACTOR (GRF). In the study, CsGIF1 in the leaf of C. sinensis was cloned and identified. The results showed that length of CsGIF1 were 666 bp, encoding 221 amino acids, and it has a highly conserved region called SNH domain. Protein analysis suggested that the CsGIF1 was hydrophily protein, and its theoretical relative molecular weight is 23,380, theoretical isoelectric point is 6.30, percentage of acidic, basic, aromatic and aliphatic amino acids is 7%, 9%, 5% and 13%. 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. The expression profiles of CsGIF1 gene were higher in tender leaves and consistently downregulated during tea leaf development. Those results suggested that CsGIF1 may be actively involved in the early stage of leaf tissue formation in tea plant. The pergence of CsGIF1 in response to different hormonal stimuli revealed the possible multiple functions of these genes in hormonal regulation. This study provided the molecular basis of the CsGIF1 gene for future functional research on leaf development and hormonal stimuli in C. sinensis. Key words: Camellia sinensis；CsGIF1；Leaf development；Hormonal stimuli 目 录 摘要1 关键词1 Abstract1 Key words1 引言1 1 材料与方法2 1.1 材料及处理 2 1.2 方法 3 1.2.1 总RNA的提取及cDNA的合成3 1.2.2 茶树CsGIF1基因的克隆3 1.2.3 序列分析4 1.2.4 定时定量PCR4 2 结果与分析4 2.1茶树CsGIF1基因的克隆4 2.2茶树CsGIF1基因编码氨基酸序列与不同物种的GIF蛋白氨基酸序列同源性比较分析5 2.3 系统进化分析6 2.4茶树CsGIF1预测蛋白序列同其他物种的蛋白质理化特性分析6 2.5茶树CsGIF1预测蛋白氨基酸序列亲水性/疏水性分析7 2.6 茶树CsGIF1转录因子的二级结构预测与分析8 2.7茶树叶片发育过程中CsGIF1基因的表达8 2.8不同植物激素处理下CsGIF1基因的表达8 3 讨论9 致谢10 参考文献10 图1 不同发育阶段茶树叶片3 (责任编辑：qin) |