拟南芥atuf2表达模式与亚细胞定位的研究.doc
约25页DOC格式手机打开展开
拟南芥atuf2表达模式与亚细胞定位的研究,拟南芥atuf2表达模式与亚细胞定位的研究摘要：非生物逆境（包括高温、低温、干旱、盐碱等）影响植物的生长发育，是制约农作物产量和质量提高的主要因素之一。鉴定与植物耐受非生物逆境相关的基因的功能，可以为提高作物的耐受非生物胁迫的能力提供理论指导。以前的研究发现拟南芥atuf2基因的表达受非生物胁迫的诱导，可能参与了拟南芥...
内容介绍
此文档由会员 小花仙66 发布
拟南芥AtUF2表达模式与亚细胞定位的研究
摘要：非生物逆境（包括高温、低温、干旱、盐碱等）影响植物的生长发育，是制约农作物产量和质量提高的主要因素之一。鉴定与植物耐受非生物逆境相关的基因的功能，可以为提高作物的耐受非生物胁迫的能力提供理论指导。以前的研究发现拟南芥AtUF2基因的表达受非生物胁迫的诱导，可能参与了拟南芥对非生物胁迫的响应。为了研究AtUF2的作用机制，我们对AtUF2基因亚细胞定位及组织表达进行了研究。研究发现，AtUF2-GFP信号定位在细胞质和细胞核中。对pAtUF2::GUS转基因植株进行GUS染色，发现，AtUF2基因在幼苗的中表达，GUS信号主要集中在子叶和下胚轴的维管组织以及根中，但是不在根尖中表达。
关键词：拟南芥；非生物胁迫；GUS；PCR
The expression pattern and subcellular localization of Arabidopsis AtUF2
Abstract: Plants meet with many adverse conditions during their growth processes. These stresses cause damage to plants in different ways. Researches on the different functions of genes which are involved in the stress-resisting process can provide new clues for improving plant tolerance to abiotic stresses. In the former experiments we find that the expression of AtUF2 induced by the abolic stress and it may be related with the reaction of abolic stress of Arabidopsis. In order to study the mechablism of AtUF2 , we focus on its expression pattern and subcellular localization. We find that the AtUF2-GFP signal is located in cytoplasm and nucleus. And the GUS results of the pAtUF2::GUS plants read that AtUF2 expresses in the seedings. The GUS signals are focused in cotyledon, hypocotyl fibrovascular tissue and root excepet the edge of root.
Key words: Arabidopsis，abiotic stress，GUS，PCR
目 录
摘要 1
关键词 1
1 前言 3
1.1 模式植株——拟南芥简介 3
1.2 非生物胁迫 3
1.2.1盐胁迫 4
1.2.2 干旱胁迫 5
1.2.3 氧化胁迫 6
1.3 GUS基因 6
2 实验材料与方法 7
2.1 实验材料 7
2.1.1 植物材料及生长条件 7
2.1.2 培养基 7
2.1.3 菌株与质粒 7
2.1.4 酶及试剂 7
2.1.5 实验仪器 8
2.2 实验方法 8
2.2.1 拟南芥的种子处理 8
2.2.2 拟南芥转化 9
2.2.3 转基因拟南芥的鉴定 9
2.2.4 AtUF2基因的显微观察 12
2.2.5 GUS组织化学染色分析 12
3 结果与分析 13
3.1 拟南芥的侵染 13
3.2 已侵染拟南芥的筛选 13
3.3 已筛拟南芥的显微观察 14
3.4 已侵染拟南芥的鉴定 14
3.5 GUS染色观察 15
4 讨论 16
4.1 关于模式植物拟南芥 16
4.2 农杆菌介导法的植物基因转化机制以及其优点和局限性 16
5结论 18
参考文献 19
致谢 21
Contents
Abstract 1
Key words 1
1 Introduction 3
1.1 Model plant Arabidopsis introduction 3
1.2 Abolic stress 3
1.2.1 Salt stress 4
1.2.2 Drought stress 5
1.2.3 Oxidative stress 6
1.3 GUS gene 6
2 Materials and methods 7
2.1 Materials 7
2.1.1 Plant material and growth conditions 7
2.1.2 Medium 7
2.1.3 Bacterial strains and plasmids 7
2.1.4 Enzymes and reagents 7
2.1.5 Laboratory instruments 8
2.2 Experimental methods 8
2.2.1 Arabidopsis seed treatment 8
2.2.2 Arabidopsis transformation 9
2.2.3 Identification of transgenic Arabidopsis 9
2.2.4 Microscopic observation of AtUF2 genes 12
2.2.5 Staining analysis of GUS tissue 12
3 Results and analysis 13
3.1 Arabidopsis infection 13
3.2 Selection of infested Arabidopsis 13
3.3 Microscopic observation of selected Arabidopsis 14
3.4 Identification of infested Arabidopsis 14
3.5 Staining of GUS tissue 15
4 Discuss 16
4.1 The model plant Arabidopsis thaliana 16
4.2 Plant genetic transformation of Agrobacterium-mediated mechanism as well as its advantages and limitations 16
5 Conclusion 18
References 19
Acknowledgements 21
摘要：非生物逆境（包括高温、低温、干旱、盐碱等）影响植物的生长发育，是制约农作物产量和质量提高的主要因素之一。鉴定与植物耐受非生物逆境相关的基因的功能，可以为提高作物的耐受非生物胁迫的能力提供理论指导。以前的研究发现拟南芥AtUF2基因的表达受非生物胁迫的诱导，可能参与了拟南芥对非生物胁迫的响应。为了研究AtUF2的作用机制，我们对AtUF2基因亚细胞定位及组织表达进行了研究。研究发现，AtUF2-GFP信号定位在细胞质和细胞核中。对pAtUF2::GUS转基因植株进行GUS染色，发现，AtUF2基因在幼苗的中表达，GUS信号主要集中在子叶和下胚轴的维管组织以及根中，但是不在根尖中表达。
关键词：拟南芥；非生物胁迫；GUS；PCR
The expression pattern and subcellular localization of Arabidopsis AtUF2
Abstract: Plants meet with many adverse conditions during their growth processes. These stresses cause damage to plants in different ways. Researches on the different functions of genes which are involved in the stress-resisting process can provide new clues for improving plant tolerance to abiotic stresses. In the former experiments we find that the expression of AtUF2 induced by the abolic stress and it may be related with the reaction of abolic stress of Arabidopsis. In order to study the mechablism of AtUF2 , we focus on its expression pattern and subcellular localization. We find that the AtUF2-GFP signal is located in cytoplasm and nucleus. And the GUS results of the pAtUF2::GUS plants read that AtUF2 expresses in the seedings. The GUS signals are focused in cotyledon, hypocotyl fibrovascular tissue and root excepet the edge of root.
Key words: Arabidopsis，abiotic stress，GUS，PCR
目 录
摘要 1
关键词 1
1 前言 3
1.1 模式植株——拟南芥简介 3
1.2 非生物胁迫 3
1.2.1盐胁迫 4
1.2.2 干旱胁迫 5
1.2.3 氧化胁迫 6
1.3 GUS基因 6
2 实验材料与方法 7
2.1 实验材料 7
2.1.1 植物材料及生长条件 7
2.1.2 培养基 7
2.1.3 菌株与质粒 7
2.1.4 酶及试剂 7
2.1.5 实验仪器 8
2.2 实验方法 8
2.2.1 拟南芥的种子处理 8
2.2.2 拟南芥转化 9
2.2.3 转基因拟南芥的鉴定 9
2.2.4 AtUF2基因的显微观察 12
2.2.5 GUS组织化学染色分析 12
3 结果与分析 13
3.1 拟南芥的侵染 13
3.2 已侵染拟南芥的筛选 13
3.3 已筛拟南芥的显微观察 14
3.4 已侵染拟南芥的鉴定 14
3.5 GUS染色观察 15
4 讨论 16
4.1 关于模式植物拟南芥 16
4.2 农杆菌介导法的植物基因转化机制以及其优点和局限性 16
5结论 18
参考文献 19
致谢 21
Contents
Abstract 1
Key words 1
1 Introduction 3
1.1 Model plant Arabidopsis introduction 3
1.2 Abolic stress 3
1.2.1 Salt stress 4
1.2.2 Drought stress 5
1.2.3 Oxidative stress 6
1.3 GUS gene 6
2 Materials and methods 7
2.1 Materials 7
2.1.1 Plant material and growth conditions 7
2.1.2 Medium 7
2.1.3 Bacterial strains and plasmids 7
2.1.4 Enzymes and reagents 7
2.1.5 Laboratory instruments 8
2.2 Experimental methods 8
2.2.1 Arabidopsis seed treatment 8
2.2.2 Arabidopsis transformation 9
2.2.3 Identification of transgenic Arabidopsis 9
2.2.4 Microscopic observation of AtUF2 genes 12
2.2.5 Staining analysis of GUS tissue 12
3 Results and analysis 13
3.1 Arabidopsis infection 13
3.2 Selection of infested Arabidopsis 13
3.3 Microscopic observation of selected Arabidopsis 14
3.4 Identification of infested Arabidopsis 14
3.5 Staining of GUS tissue 15
4 Discuss 16
4.1 The model plant Arabidopsis thaliana 16
4.2 Plant genetic transformation of Agrobacterium-mediated mechanism as well as its advantages and limitations 16
5 Conclusion 18
References 19
Acknowledgements 21
