| 摘要 | 第8-11页 |
| Summary | 第11-13页 |
| Abbreviations | 第14-15页 |
| Chapter1 Introduction | 第15-31页 |
| 1.1 The origin and distribution of potato | 第15页 |
| 1.2 Potato pigments | 第15-19页 |
| 1.2.1 Anthocyanins - one of the major plant pigments | 第15-18页 |
| 1.2.2 Anthocyanins in potato and human health | 第18-19页 |
| 1.3 The anthocyanin biosynthetic pathway | 第19-20页 |
| 1.4 Transcription factors | 第20-21页 |
| 1.5 Regulation of anthocyanin biosynthesis | 第21-30页 |
| 1.5.1 MYB TFs involved in anthocyanin biosynthesis | 第21-26页 |
| 1.5.1.1 Activators | 第21-25页 |
| 1.5.1.2 Repressors | 第25-26页 |
| 1.5.2 Regulation of anthocyanin biosynthesis by transcriptional factor complex | 第26-30页 |
| 1.6 Aim of this thesis | 第30-31页 |
| Chapter 2 Comparative transcriptome analysis of white and purple potato to identify genesinvolved in anthocyanin biosynthesis | 第31-80页 |
| 2.1 Materials and methods | 第32-39页 |
| 2.1.1 Plant material, RNA extraction, library construction and RNA sequencing | 第32-33页 |
| 2.1.2 RNA Data filtering and de novo transcriptome assembly | 第33-34页 |
| 2.1.3 Differentially gene expression test | 第34页 |
| 2.1.4 Annotation and predicted CDS | 第34-35页 |
| 2.1.5 Gene ontology functional enrichment and pathway analysis of DEGs | 第35页 |
| 2.1.6 Quantitative real-time PCR (q PCR) analysis | 第35-36页 |
| 2.1.7 SNPs and in Del discovery | 第36-37页 |
| 2.1.8 SNPs validation and expression analysis of UFGT in tetraploid cultivars | 第37-39页 |
| 2.2 Results | 第39-75页 |
| 2.2.1 Total RNA sequencing and de novo transcriptome assembly | 第39-41页 |
| 2.2.2 Analysis, functional annotation and KEGG classification of differentially expressedgenes | 第41-45页 |
| 2.2.3 Detection of genes related to anthocyanin biosynthesis pathway | 第45-64页 |
| 2.2.4 Confirmation of differentially expressed genes by q PCR | 第64-68页 |
| 2.2.5 SNPs and in Del discovery in MYB AN1, b HLH1 and UFGT genes | 第68-75页 |
| 2.3 Discussion | 第75-79页 |
| 2.4 Conclusions | 第79-80页 |
| Chapter 3 The MYB transcription factor St MYBA1 from potato requires light to activateanthocyanin biosynthesis in transgenic tobacco | 第80-95页 |
| 3.1 Materials and methods | 第81-85页 |
| 3.1.1 Plant materials | 第81页 |
| 3.1.2 RNA, DNA isolation and first-strand c DNA synthesis | 第81页 |
| 3.1.3 Gene cloning and sequence analysis | 第81-82页 |
| 3.1.4 Electroporation of St MYBA1 construct and promoter analysis | 第82页 |
| 3.1.5 Transient assays of gene function | 第82-83页 |
| 3.1.6 Tobacco transformation | 第83页 |
| 3.1.7 Quantitive real-time PCR (q PCR) analysis of gene expression | 第83-84页 |
| 3.1.8 HPLC analysis of transgenic tobacco plants | 第84-85页 |
| 3.2 Results | 第85-93页 |
| 3.2.1 Functional assays of St MYBA1-1 and St MYBA1-2 in tobacco | 第85页 |
| 3.2.2 Phenotype of T0 transgenic plants | 第85-86页 |
| 3.2.3 St MYBA1 activates most of the anthocyanin pathway genes in leaves and roots oftransformed tobacco lines | 第86-88页 |
| 3.2.4 Over-expression of St MYBA1 activates the expression of endogenous b HLHs intransgenic tobacco | 第88-89页 |
| 3.2.5 Anthocyanin biosynthesis in tobacco regulated by St MYBA1 was affected by light | 第89-92页 |
| 3.2.6 St MYBA1 upstream promoter region sequence analysis | 第92-93页 |
| 3.3 Discussion | 第93-95页 |
| Chapter 4 Functional diversification of the potato R2R3 MYB anthocyanin activators AN1, MYBA1, and MYB113 and their interaction with basic helix-loop-helix cofactors | 第95-140页 |
| 4.1 Materials and methods | 第96-108页 |
| 4.1.1 Plant materials | 第96-97页 |
| 4.1.2 Determination of anthocyanin content of potato skin and flesh | 第97-98页 |
| 4.1.3 DNA and RNA extraction | 第98页 |
| 4.1.4 Gene cloning and sequence analysis | 第98-105页 |
| 4.1.5 Transient assays of gene function | 第105-106页 |
| 4.1.6 Phylogenetic analysis | 第106页 |
| 4.1.7 Transformation of tobacco | 第106页 |
| 4.1.8 HPLC measurement of tobacco leaves | 第106-107页 |
| 4.1.9 Quantitative real-time PCR | 第107页 |
| 4.1.10 R repeat function verification | 第107-108页 |
| 4.1.11 Statistical analysis | 第108页 |
| 4.2 Results | 第108-135页 |
| 4.2.1 Tuber anthocyanin content in four potato genotypes | 第108页 |
| 4.2.2 Analysis of St AN1, St MYBA1, and St MYB113 gene sequences in differentiallypigmented cultivars | 第108-115页 |
| 4.2.3 q PCR analysis of St AN1, St MYBA1, and St MYB113 in four potato genotypes | 第115-117页 |
| 4.2.4 Analysis of Stb HLH gene sequences in differentially pigmented cultivars | 第117-122页 |
| 4.2.5 q PCR analysis of Stb HLHs in four potato genotypes | 第122-123页 |
| 4.2.6 Functional assays of St AN1, St MYBA1 and St MYB113 in tobacco | 第123-127页 |
| 4.2.7 St AN1-R0, St AN1-R1, and St AN1-R3 activate all the anthocyanin biosyntheticgenes in leaves and roots of transformed tobacco lines | 第127-131页 |
| 4.2.8 St AN1, St MYBA1, and St MYB113 partner with Stb HLH1 and St JAF13 to regulateanthocyanin biosynthesis | 第131-135页 |
| 4.3 Discussion | 第135-138页 |
| 4.3.1 Three major variants of St AN1 present in four different pigmented cultivars | 第136页 |
| 4.3.2 The involvement of St AN1-R0, St AN1-R1, and St AN1-R3 in regulating anthocyaninbiosynthesis | 第136-137页 |
| 4.3.3 StbHLH1 and StJAF13 are limiting regulators of anthocyanin biosynthesis in potatotubers | 第137-138页 |
| 4.4 Conclusion | 第138-140页 |
| References | 第140-155页 |
| 致谢 | 第155-157页 |
| 作者简介 | 第157-159页 |
| 导师简介 | 第159-161页 |
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