拟南芥SEC6基因在主根细胞程序性死亡中的功能初步研究
时间:2018-07-17 20:53 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：Exocyst复合体作为引导囊泡与靶膜结合的拴系过程的拴系因子，在真核生物中普遍存在。目前已知在模式植物拟南芥中，exocyst复合体参与细胞板形成、生长素运输、细胞极性生长、细胞自噬等多种重要的生物学过程。细胞的程序性死亡（Programmed Cell Death，PCD）对植物体的生长发育、形态构成、生物防御等方面具有重要作用。前期实验中我们发现，SEC6基因的突变会导致拟南芥主根细胞发生 PCD 现象。为更加深入地了解 exocyst 复合体亚基 SEC6 基因突变导致拟南芥主根细胞发生PCD的分子机制，我们需要做更加深入的研究。 本实验中，我们以exocyst复合体亚基 SEC6基因的花粉回补突变体 PRsec6-1 为材料，并通过杂交获得 rbohD 和 ein2-5 背景下的 PRsec6-1 突变体，通过台盼蓝、PI、DCFH-DA、TUNEL等染料染色,分析其主根发育表型。结果发现，阻断RboHD产生的活性氧信号途径不能缓解主根细胞 PCD 的表型，而阻断乙烯信号途径可缓解该 PCD 表型。说明exocyst复合体亚基SEC6基因突变导致的主根细胞程序性死亡依赖于乙烯信号途径。25790 毕业论文关键词：Exocyst复合体；SEC6；主根；细胞程序性死亡；乙烯信号途径 Preliminary Research of SEC6 Gene Function in Programmed Cell Death of Arabidopsis Primary Root Abstract： Exocyst complex is a tethering factor that directs the tethered process of the vesicle to the target membrane, which is prevalent in eukaryotes. It is known that exocyst complex participates in a variety of important biological processes such as cell plate formation, auxin transport, cell polarity growth and cell autophagy in model plant Arabidopsis. Programmed Cell Death (PCD) plays an important role in the growth and development, morphological composition and biological defense of plants. In the previous experiment, we found that mutations in the SEC6 gene lead to PCD in Arabidopsis thaliana root cells. To understand more deeply the molecular mechanism of PCD in Arabidopsis thaliana root cells by mutations in the exocyst complex subunit gene SEC6, we need to do more in-depth study. In this study, we used the exocyst complex subunit SEC6 gene pollen-compensating mutant PRsec6-1 as a material, and obtain double mutant PRsec6-1 rbohD and PRsec6-1 ein2-5 by hybridization, then dye with trypan blue, PI , DCFH-DA, TUNNEL to analysis the development phenotype of the primary root. We found that blocking the reactive oxygen species pathway did not alleviate the phenotype of PCD in the main root cells, while blocking the ethylene signaling pathway could alleviate the PCD phenotype. Indicating that the primary root cell programmed death caused by gene mutate of exocyst complex subunit SEC6 may depend on the ethylene signaling pathway. Key words: Exocyst complex；SEC6；Primary Root；Programmed cell death；Ethylene signal pathway 目 录 摘要3 关键词3 Abstract3 Key words3 引言（或绪论）3 1 材料与方法4 1.1 实验材料 4 1.1.1 植物材料4 1.1.2 酶、试剂及各种化学药品4 1.2 实验仪器 5 1.3 实验方法 5 1.3.1 拟南芥的栽培与管理5 1.3.2 拟南芥总 DNA 的快速提取（Edward 法）5 1.3.3 引物设计及 PCR 扩增6 1.3.4 拟南芥的人工杂交6 1.3.5 主根表型观察6 1.3.6 细胞死亡及细胞程序性死亡（PCD）检测6 (责任编辑：qin) |