List of Abbreviations | 第10-12页 |
摘要 | 第12-15页 |
Abstract | 第15-17页 |
Chapter 1 Introduction | 第18-34页 |
1.1 Flavonoid biosynthesis pathway | 第19-24页 |
1.1.1 Anthocyanin | 第21页 |
1.1.2 Proanthocyanin | 第21-22页 |
1.1.3 Flavonol | 第22-23页 |
1.1.4 Phenolics Acid | 第23-24页 |
1.2 MYB genes as transcription factors | 第24-28页 |
1.2.1 MYB transcription factor | 第24-25页 |
1.2.2 Structure of MYB | 第25-26页 |
1.2.3 Evolution of MYB | 第26页 |
1.2.4 Tissue-specific expression | 第26-27页 |
1.2.5 Biological Functions of MYB | 第27-28页 |
1.3 Regulation of flavonoid biosynthesis | 第28-29页 |
1.4 MYB repressors | 第29-32页 |
1.5 The genetic transformation in Chinese narcissus | 第32-33页 |
1.6 The objectives of study | 第33-34页 |
Chapter 2 Materials and Methods | 第34-44页 |
2.1 Plant materials | 第34页 |
2.2 RNA extraction and cDNA synthesis | 第34-35页 |
2.3 3-RACE of NtMYB2 | 第35-36页 |
2.4 Cloning of NtMYB3 | 第36-37页 |
2.5 Cloning of NtMYB4 | 第37-38页 |
2.6 Expression Vector Construction | 第38-39页 |
2.7 Sequence analysis of NtMYB2, NtMYB3 and NtMYB4 | 第39页 |
2.8 The expression pattern of NtMYB2, NtMYB3 and NtMYB4 in different tissues of ChineseNarcissus | 第39-40页 |
2.9 Transient assays of NtMYB2, NtMYB3 and NtMYB4 gene function | 第40-41页 |
2.10 Stable transformation of tobacco | 第41页 |
2.11 Extraction and measurement of total anthocyanins and phenolics | 第41-42页 |
2.12 Statistical Analysis | 第42-44页 |
Chapter 3 Identification and function characterization of a R2R3-MYB, NtMYB2 in ChineseNarcissus | 第44-58页 |
3.1 Results | 第44-53页 |
3.1.1 Cloning of NtMYB2 | 第44页 |
3.1.2 Subcellular localization | 第44页 |
3.1.3 Sequence Analysis and Phylogenetic tree | 第44-48页 |
3.1.4 The expression pattern of NtMYB2 in Chinese Narcissus | 第48页 |
3.1.5 Transient expression of NtMYB2 in Tobacco | 第48-49页 |
3.1.6 Stable expression of MtMYB2 in tobacco | 第49-51页 |
3.1.7 Overexpression of NtMYB2 in tobacco influences the expression levels of flavonoidbiosynthetic pathway genes | 第51-53页 |
3.2 Discussion | 第53-58页 |
3.2.1 Bioinformatics analysis showed NtMYB2 was a transcriptional factor | 第54-55页 |
3.2.2 NtMYB2 reduced the pigmentation in tobacco leaves with transient assay | 第55页 |
3.2.3 NtMYB2 reduces the flower color of transgenic tobacco | 第55-56页 |
3.2.4 NtMYB2 is involved in regulation of anthocyanin /flavonoid biosynthesis pathway genes intobacco | 第56-57页 |
3.2.5 Expression Profiles of NtMYB2 in narcissus | 第57-58页 |
Chapter 4 Identification and functional characterization a R2R3-MYB, NtMYB3 in Chinesenarcissus | 第58-73页 |
4.1 Results | 第58-67页 |
4.1.1 Subcellular localization | 第58页 |
4.1.2 Sequence Analysis and Phylogenetic tree | 第58-59页 |
4.1.3 The expression pattern of NtMYB3 in Chinese Narcissus | 第59-62页 |
4.1.4 Transient expression of NtMYB3 in Tobacco | 第62-64页 |
4.1.5 The overexpression of NtMYB3 decreased the pigmentation in the flowers of transgenictobacco lines | 第64-65页 |
4.1.6 Ectopic expression of NtMYB3 affects expression of key genes for anthocyanin and PAbiosynthesis in transgenic tobacco | 第65-67页 |
4.2 Discussion | 第67-73页 |
4.2.1 NtMYB3 could reduce flavonoid biosynthesis through the C2 repressor/ EAR motif | 第67-69页 |
4.2.2 Expression pattern of the NtMYB3 in different tissues of Chinese narcissus | 第69页 |
4.2.3 NtMYB3 decreased the anthocyanin in agroinfiltrated tobacco leaves | 第69-70页 |
4.2.4 NtMYB2 change the flower color of transgenic tobacco | 第70-71页 |
4.2.5 NtMYB3 negatively regulate anthocyanin and flavonol biosynthesis pathway genes intransgenic tobacco | 第71-72页 |
4.2.6 NtMYB3 positively regulate proanthocyanidin biosynthesis pathway genes in transgenictobacco | 第72页 |
4.2.7 Total Phenolics contents | 第72-73页 |
Chapter 5 Overexpression of Chinese narcissus a R2R3-MYB transcriptional factor, NtMYB4,reduced the flower pigmentation and caused the dwarfism in transgenic tobacco | 第73-88页 |
5.1 Results | 第73-84页 |
5.1.1 Characterization of NtMYB4 as a transcriptional factor | 第73-74页 |
5.1.2 Subcellular localization | 第74页 |
5.1.3 NtMYB4 is a flower specific gene | 第74-77页 |
5.1.4 Transient expression of NtMYB4 reduced pigmentation in tobacco leaves | 第77-79页 |
5.1.5 Transient expression of NtMYB4 changes the expression pattern of anthocyaninbiosynthesis gene in tobacco leaves | 第79页 |
5.1.6 Overexpression of NtMYB4 alter the pigmentation pattern in transgenic flowers | 第79-82页 |
5.1.7 NtMYB4 affects expression of flavonoid genes in transgenic tobacco flowers | 第82-84页 |
5.3 Discussion | 第84-88页 |
5.3.1 Bioinformatics analysis | 第84页 |
5.3.2 Expression pattern of NtMYB4 in different tissues | 第84-85页 |
5.3.3 Ectopic overexpression of NtMYB4 in tobacco | 第85-86页 |
5.3.4 The expression analysis of endogenous anthocyanin/flavonoid biosynthetic genes intobacco flowers | 第86-88页 |
Chapter 6 Conclusions and prospects | 第88-89页 |
References | 第89-96页 |
Published Papers during the Programme | 第96-97页 |
Acknowledgements | 第97-98页 |
Supplementary Materials | 第98-101页 |