| 摘要 | 第7-9页 |
| ABSTRACT | 第9-10页 |
| LIST OF ABBREVIATIONS | 第11-12页 |
| CHAPTER Ⅰ. REVIEW OF LITERATURE | 第12-45页 |
| 1.0 Background | 第12页 |
| 1.1 Soybean: A source of worldwide food with special type of metabolites | 第12页 |
| 1.2 Isoflavonoids:specialized legume metabolites | 第12-19页 |
| 1.2.1 Isoflavonoids:vital role in human health and important element of daily diet | 第13-14页 |
| 1.2.2 Isoflavonoids:competent role in plant-environment interaction | 第14-19页 |
| 1.3 Isoflavonoids:structural diversity, accumulation, and synthesis in differentsoybean tissues | 第19-25页 |
| 1.3.1 Isoflavonoids: Diversity in structure | 第19-21页 |
| 1.3.2 Isoflavonoids accumulation in soybean | 第21-23页 |
| 1.3.3 Isoflavonoids:Biosynthesis pathway | 第23-25页 |
| 1.4 Isoflavonoid under acylation process | 第25-44页 |
| 1.4.1 BAHD related MTs in specific species | 第26-28页 |
| 1.4.2 MTs are available in different plant tissues | 第28-31页 |
| 1.4.3 Specificity of MTs towards their substrates | 第31-36页 |
| 1.4.4 Structural characterization of MTs | 第36-37页 |
| 1.4.5 Phylogeny relationship of BAHD MT enzymes | 第37-40页 |
| 1.4.6 Application of MTs | 第40-44页 |
| 1.5 Objective of the study | 第44-45页 |
| CHAPTER Ⅱ. MATERIALS AND METHODS | 第45-57页 |
| 2.1 Identification of the soybean IMaT gene family and in silico analysis | 第45页 |
| 2.2 Phylogenetic analysis | 第45页 |
| 2.3 RNAseq expression profile | 第45-46页 |
| 2.4 Cloning of malonyltransferase genes | 第46-48页 |
| 2.5 Protein induction and activity assay | 第48-50页 |
| 2.5.1 Heterologous expression of recombinant GmIMaTs | 第48-49页 |
| 2.5.2 Enzyme assay and HPLC analyses | 第49-50页 |
| 2.5.3 Kinetic analysis | 第50页 |
| 2.6 Gene expression analysis using quantitative reverse transcriptase (qRT)-PCR andIsoflavone quantification in different parts | 第50-51页 |
| 2.7 Overexpression and RNAi knockdown of GmIMaT1 and GmIMaT3 in soybeanhairy roots | 第51-52页 |
| 2.7.1 Extraction and analysis of isoflavonoids from Soybean hairy roots | 第52页 |
| 2.8 Plasmid construction and Subcellular localization of GmIMaT1 and GmIMaT3 | 第52-53页 |
| 2.9 Soybean plant treatment for gene expression and isoflavone metabolite analyses | 第53-54页 |
| 2.10 In-vivo hairy root and induction | 第54-55页 |
| 2.10.1 Generation of GmIMaTs-overexpression and -knockdown chimerical soybeanplants for nodulation assay | 第54-55页 |
| 2.10.2 Effect of B. japonicum on GmIMaTs expression in early stage | 第55页 |
| 2.10.3 Expression level of GmIMaTs and isoflavone contents in different noduledeveloping stages | 第55页 |
| 2.10.4 B. japonicum affect the expression of GmIMaTs in overexpressed andknockdown chimeric plants | 第55页 |
| 2.11 Bioinformatics analysis | 第55-56页 |
| 2.12 Statistic analysis | 第56-57页 |
| CHAPTER Ⅲ. RESULTS | 第57-104页 |
| 3.1 cDNA cloning, Sequence analysis and phylogeny of GmIMaTl, GmIMaT2,GmIMaT3 and GmIMaT4 | 第57-63页 |
| 3.2 Transcription analyses | 第63-69页 |
| 3.3 Recombinant protein production and enzyme activity assay | 第69-76页 |
| 3.4 Overexpression and silencing of GmIMaTs altered isoflavone profiles in hairyroots | 第76-82页 |
| 3.5 Subcellular localization of GmIMaT1 and GmIMaT3 | 第82-83页 |
| 3.6 GmIMaT1 and GmIMaT3 are differently regulated by various environmental andhormonal factors | 第83-90页 |
| 3.6.1 GmIMaT1 and GmIMaT3 expression and isoflavone content in response to cold(4 ℃) treated seeds | 第83-84页 |
| 3.6.2 GmIMaT1 and GmIMaT3 expression and isoflavone content in response to heat(42 ℃) treated seeds and leaves | 第84-86页 |
| 3.6.3 GmIMaT1 and GmIMaT3 expression and isoflavone content in response todrought, low pH and AL3+ treated roots | 第86-88页 |
| 3.6.4 GmIMaT1 and GmIMaT3 expression and isoflavone content in response tohormonal (ABA and MeJA) stress | 第88-90页 |
| 3.7 Role of GmIMaTs in nodulation | 第90-104页 |
| 3.7.1 Expression level and isoflavone contents in different nodule developing stages | 第90-93页 |
| 3.7.2 Overexpression or Knockdown of GmIMaT1 and GmIMaT3 in transgenic hairyroots altered soybean nodulation | 第93-100页 |
| 3.7.3 Transcript response of GmIMaT1 and GmIMaT3 upon B. japonicum infectionin wild type seedlings | 第100-102页 |
| 3.7.4 GmIMaT1 and GmIMaT3 overexpression and knockdown hairy roots responseto B. japonicum infection in chimeric plants | 第102-104页 |
| CHAPTER Ⅳ. DISCUSSION | 第104-110页 |
| 4.1 GmIMaT1 and GmIMaT3 catalyze the malonylation of isoflavone glucosides invitro | 第104-106页 |
| 4.2 Subcellular and tissue specific expression of GmIMaTs for isoflavonemodification | 第106页 |
| 4.3 GmIMaTs are responsible for the malonylation of soybean isoflavone glucosides | 第106-107页 |
| 4.4 GmIMaT1 and GmIMaT3 differentially respond to abiotic,biotic and hormonalstresses | 第107-108页 |
| 4.5 GmIMaTs dependent isoflavone essential for nodulation in soybean | 第108-110页 |
| CONCLUSION | 第110-111页 |
| REFERENCES | 第111-128页 |
| ACKNOWLEDGEMENT | 第128-130页 |
| PUBLICATIONS | 第130页 |
