学位专题

封面
声明
中文摘要
英文摘要
目录
主要符号对照表/ABBREVIATIONS
1.1 INTRODUCTION
1.2 JUSTIFICATION
1.3 OBJECTIVES
1.4 APOCYNUM SPECIES–Apocynum venetum and Apocynum hendersonii
1.4.1. Propagation
1.4.2Constituents’ chemicalsin Apocynum
1.4.3Economic importance of Apocynum
1.4.4Flavonoids,types and functions
1.4.5. Flavonoid biosynthesis pathways
CHAPTER2:GENOME ASSEMBLY AND CHARACTERIZATION OF FLAVONOID BIOSYNTHESIS GENES IN APOCYNUM
2.1 INTRODUCTION
2.2MATERIALS AND METHODS
2.2.1Genomic DNA extraction and sequencing
2.2.2Hi-C library and sequencing
2.2.3 Genome assembly and evaluation
2.2.4 Genome annotation
2.2.5 Genome repeat sequences identification
2.2.6Evolutionary and synteny analysis
2.3RESULTS AND DISCUSSION
2.3.1Sequencing and high-quality assembly of A. venetum and A. hendersonii genomes
2.3.2Genome annotation and comparative genomic analysis
2.3.3 Repetitive sequences and Transposable elements analysis
2.3.4Gene structure and genome whole duplication event
2.3.5Characterization of putative flavonoid biosynthesis-related genes
3.1 INTRODUCTION
3.2MATERIALS AND METHODS
3.2.1Identification of MYB protein fromApocynumspecies
3.2.2Chromosome distribution, geneduplication and synteny analysis
3.2.3Structural classification and Motif analysis of the MYBs
3.2.4Identification of cis-elements
3.2.5RNA-seq acquisition and analysis
3.3RESULTS AND DISCUSSIONS
3.3.1Identification of MYB proteinsfromApocynum species
3.3.2Chromosome distribution, gene duplication and synteny analysis
3.3.3Phylogenetic divergence of the AvR2R3-MYB genes
3.3.4Structural classification and Motif analysis of the AvR2R3-MYBs
3.3.5Cis-elements in the AvR2R3-MYBs
3.3.6RNA-seq data analysis and expression pattern of Av/Av-R2R3-MYBsin different tissues
3.3.7Relative expression profile of anthocyanin biosynthesis candidate MYB genes in the the two Apocynum species
4.1 INTRODUCTION
4.2MATERIAL AND METHODS
4.2.1Construction of F3H Over-expression Vector
4.2.2 Plant materials
4.2.3 Primer design
4.2.4 RNA extraction and cDNA synthesis
4.2.5 PCR amplification, gene cloning and sequence analysis
4.2.6 Transformation ofA. venetumandA. hendersoniiusing Agrobacterium carrying the F3H gene
4.2.7 Transformation of the modal plant, Arabidopsis using Agrobacterium carrying the F3H gene
4.2.8 Treatment
4.2.9 Total flavonoids, total tannins and anthocyanin contents
4.3RESULTS AND DISCUSSION
4.3.1Physico-chemical properties of the F3H
4.3.2 Cloning, sequence analysis and transformation
4.3.3 Flavonoids contents,proanthocyanidin, and anthocyaninscontents in the transgenicA. thalianaharbouring theF3H-pBI121 construct
CHAPTER5:UDP-GLUCOSE: FLAVONOID 3-O-GLUCOSYLTRANSFERASES (UGTs) GENES CLONING, PROKRAYOTIC EXPRESSION AND ENZYMATIC ASSAYS
5.1 INTRODUCTION
5.2MATERIALS AND METHODS
5.2.1Construction ofUGTsExpression Vector
5.2.2Plant materials
5.2.3Primer design
5.2.4RNA extraction,cDNA synthesis, amplification and cloning
5.2.5Expression analysis
5.2.6 Enzymatic assay and substrate specificity test
5.2.7 Enzyme Kinetics
5.3RESULT AND DISCUSSION
5.3.1 UGTs selection
5.3.2Physico-chemical properties and structure of the UGTs
5.3.3Phylogenetic analysis of the UGTs
5.3.4 Substrate specificity
5.3.5Enzyme kinetics
6.1GENOME ASSEMBLY AND RELATED ANALYSIS
6.2 BIOINFORMATICS ANALYSIS OF MYB TFs
6.3 F3H OVEREXPRESSION AND ANALYSIS
6.4 UGTs CLONING, EXPRESSION AND CHARACTERIZATION
6.5 CONCLUSION
6.6 PROSPECTS
参考文献
附录 (Appendix)
致谢
作者简历(CURRICULUM VITAE)