擬南芥AtUF2表達(dá)模式與亞細(xì)胞定位的研究

摘要：非生物逆境（包括高溫、低溫、干旱、鹽堿等）影響植物的生長發(fā)育，是制約農(nóng)作物產(chǎn)量和質(zhì)量提高的主要因素之一。鑒定與植物耐受非生物逆境相關(guān)的基因的功能，可以為提高作物的耐受非生物脅迫的能力提供理論指導(dǎo)。以前的研究發(fā)現(xiàn)擬南芥AtUF2基因的表達(dá)受非生物脅迫的誘導(dǎo)，可能參與了擬南芥對(duì)非生物脅迫的響應(yīng)。為了研究AtUF2的作用機(jī)制，我們對(duì)AtUF2基因亞細(xì)胞定位及組織表達(dá)進(jìn)行了研究。研究發(fā)現(xiàn)，AtUF2-GFP信號(hào)定位在細(xì)胞質(zhì)和細(xì)胞核中。對(duì)pAtUF2::GUS轉(zhuǎn)基因植株進(jìn)行GUS染色，發(fā)現(xiàn)，AtUF2基因在幼苗的中表達(dá)，GUS信號(hào)主要集中在子葉和下胚軸的維管組織以及根中，但是不在根尖中表達(dá)。
關(guān)鍵詞：擬南芥；非生物脅迫；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 錄

摘要 1
關(guān)鍵詞 1
1　前言 3
1.1 模式植株——擬南芥簡(jiǎn)介 3
1.2 非生物脅迫 3
1.2.1鹽脅迫 4
1.2.2 干旱脅迫 5
1.2.3 氧化脅迫 6
1.3 GUS基因 6
2　實(shí)驗(yàn)材料與方法 7
2.1 實(shí)驗(yàn)材料 7
2.1.1 植物材料及生長條件 7
2.1.2 培養(yǎng)基 7
2.1.3 菌株與質(zhì)粒 7
2.1.4 酶及試劑 7
2.1.5 實(shí)驗(yàn)儀器 8
2.2 實(shí)驗(yàn)方法 8
2.2.1 擬南芥的種子處理 8
2.2.2 擬南芥轉(zhuǎn)化 9
2.2.3 轉(zhuǎn)基因擬南芥的鑒定 9
2.2.4 AtUF2基因的顯微觀察 12
2.2.5 GUS組織化學(xué)染色分析 12
3 結(jié)果與分析 13
3.1 擬南芥的侵染 13
3.2 已侵染擬南芥的篩選 13
3.3 已篩擬南芥的顯微觀察 14
3.4 已侵染擬南芥的鑒定 14
3.5 GUS染色觀察 15
4 討論 16
4.1 關(guān)于模式植物擬南芥 16
4.2 農(nóng)桿菌介導(dǎo)法的植物基因轉(zhuǎn)化機(jī)制以及其優(yōu)點(diǎn)和局限性 16
5結(jié)論 18
參考文獻(xiàn) 19
致謝 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