摘要：茶树(Camellia sinensis(L.)O. Kuntze)，属山茶科山茶属，为多年生常绿木本植物，用于在世界范围内生产非酒精性饮料。GROWTH-REGULATING FACTOR (GRF)生长调节因子是植物特有的蛋白，可以积极的调节叶发育。在本研究中，对茶树转录组中的CsGRF2基因进行了克隆和鉴定。实验结果表明，茶树的CsGRF2基因全长1,068 bp，编码355个氨基酸，编码一个QLQ和一个WRC结构域。通过蛋白质分析可知，茶树CsGRF2基因的理论相对分子质量为39,110，理论等电点pI为8.34，酸性氨基酸、碱性氨基酸、芳香族氨基酸和脂肪族氨基酸比例分别为9%、14%、9%和13%。CsGRF2基因的表达水平在嫩叶中较高，随茶树叶发育过程持续下降。这些结果表明，CsGRF2基因可能积极地参与了茶树叶早期的形成。此外，响应不同激素刺激的差异表达揭示了CsGRF2基因在激素调节中的多重功能。本研究为茶树GRF家族基因对叶的生长发育和对激素处理的反应提供一个潜在的分子基础。28057
毕业论文关键词：茶树；GRF；叶发育、激素刺激
Cloning and functional identification of leaf development-related CsGRF2 genes in Camellia sinensis
Abstract：Tea plant (Camellia sinensis (L.) O. Kuntze), which belongs under the genus Camellia of the family Theaceae, is a perennial evergreen woody crop for non-alcoholic beverage production worldwide. The GROWTH-REGULATING FACTOR (GRF) is the specific protein in plants that positively regulates leaf development. In this study, CsGRF2 in the leaf transcriptome of C. sinensis was cloned and identified. The results showed the length of CsGRF2 is 1,068 bp and encodes 355 amino acids, which contains a QLQ and one WRC domains. Protein analysis suggested that: the theoretical molecular weight of the CsGRF2 predicted protein is 39,110; theoretical isoelectric point is 8.34; the percentage of acidic, basic, aromatic and aliphatic amino acids is 9%, 14%, 9% and 13%, respectively. The expression level of CsGRF2 gene is higher in the tender leaves, but expression decrease consistently during tea plant leaves development. These results suggested that the CsGRF2 gene may actively participate in the early formation of tea leaves. Besides, the differential expression of response to different hormonal stimuli reveals the multiple functions of CsGRF2 genes in hormonal regulation. This study provides a potential molecular basis of GRF family gene for researching the leaf development and hormonal stimuli in tea plant.
Key words: Tea plant; GRF; Leaf development; Hormonal stimuli
目 录
摘要 1
关键词 1
Abstract 1
Key words 1
引言 1
1 材料与方法 2
1.1实验材料与处理 2
1.2 实验方法 3
1.2.1 总RNA的提取及cDNA的合成 3
1.2.2 茶树CsGRF2基因的克隆 4
1.2.3 序列分析 4
1.2.4 实时定量PCR(qRT-PCR)检测 4
2 结果与分析 4
2.1 茶树CsGRF2基因的克隆 4
2.2 茶树CsGRF2基因编码氨基酸序列与不同物种的GRF蛋白氨基酸序列同源性比较分析 5
2.3茶树转录因子CsGRF2的系统进化树分析 7
2.4 茶树CsGRF2预测蛋白序列同其他物种的蛋白质理化特性分析 7
2.5 茶树叶发育过程中CsGRF2基因的表达 8
2.6 茶树CsGRF2基因在不同激素处理下的表达谱 9
3 讨论 9
3.1 实验结果 9
3.2 本实验存在的不足和展望 10
致谢 10
参考文献： 11
图 1 茶树‘龙井43’叶片的不同发育阶段 3 茶树发育相关CsGRF2基因克隆与功能鉴定:http://www.youerw.com/shengwu/lunwen_22767.html