除蟲(chóng)菊是一種重要的經(jīng)濟(jì)作物,以其花頭富含有高量的殺蟲(chóng)次生代謝物除蟲(chóng)菊酯著稱。除蟲(chóng)菊酯能在除蟲(chóng)菊地上部分合成,但是其中含量最高的合成器官為子房瘦果。前人證實(shí)位于除蟲(chóng)菊花和瘦果以及果皮外分泌管上的腺體組織能參與除蟲(chóng)菊酯的合成代謝。除蟲(chóng)菊酯合成途徑的第一個(gè)關(guān)鍵步驟是利用菊基焦磷酸合成酶（CDS）催化兩分子的烯丙基二磷酸（DMAPP）產(chǎn)生菊基二磷酸（CPP）。除蟲(chóng)菊TcCDS基因由FDS因家族通過(guò)基因復(fù)制進(jìn)化而來(lái),除蟲(chóng)菊TcCDS基因與菊科近緣植物青蒿體內(nèi)的FDS基因具有70%的同源性。我們首次克隆了除蟲(chóng)菊TcCDS基因啟動(dòng)子并分析其腺體組織特異性,并且分析了啟動(dòng)子序列上結(jié)合不同轉(zhuǎn)錄因子以及響應(yīng)環(huán)境和發(fā)育條件等能影響TcCDS表達(dá)和除蟲(chóng)菊酯合成的順式元件,這為將來(lái)除蟲(chóng)菊更進(jìn)一步的實(shí)驗(yàn)提供了一定的基礎(chǔ)理論。同時(shí),為利用腺體特異啟動(dòng)子所進(jìn)行的代謝工程從而提高植物次生代謝物含量和產(chǎn)量提供了幫助。在以前的研究中,通過(guò)傳統(tǒng)育種以及化學(xué)激素處理（乙烯利、矮壯素、多效唑）均能提高除蟲(chóng)菊酯的含量,但是相對(duì)于調(diào)控生物合成途徑的代謝工程具有一定的局限性。目前還未有人嘗試通過(guò)代謝工程提高除蟲(chóng)菊酯含量。在青蒿中,已有... 

【文章來(lái)源】：華中農(nóng)業(yè)大學(xué)湖北省 211工程院校 教育部直屬院校

【文章頁(yè)數(shù)】：104 頁(yè)

【學(xué)位級(jí)別】：博士

【文章目錄】：
Abstract
摘要
List of abbreviations
1 General Introduction
    1.1 Morphology and Flower development
    1.2 History and Cultivation area
    1.3 Plant grandular trichome:specialized metabolites factories
    1.4 Chemistry of pyrethrins
    1.5 Role of pyrethrins as natural insecticide
    1.6 Pyrethrins Biosynthesis
    1.7 Trichome specific promoter
    1.8 Metabolic engineering of plant secondary metabolites
    1.9 Molecular evolution and phylogeny of the nuclear gene
    1.10 Objective of the thesis
2 Molecular Clining and Characterization of the Trichome Specific ChrysanthemylDiphosphate Synthase Promoter from Tanacetum cinerariifolium
    2.1 Abstract
    2.2 Introduction
    2.3 Materials and methods
        2.3.1 Plant materials
        2.3.2 Promoter cloning
        2.3.3 DNA sequence analysis
        2.3.4 Vector construction
        2.3.5 Plant transformation
        2.3.6 Gus assay
    2.4 Results
        2.4.1 Characterization of the promoter DNA sequence
        2.4.2 Tissue specificity of the TcCDS promoter in tobacco
    2.5 Discussion
3 Overexpression of the genes Chrysanthemyl Diphosphate Synthase （TcCDS） increasedPyrethrins content in Tanacetum cinerariifolium
    3.1 Abstract
    3.2 Introduction
    3.3 Material and Methods
        3.3.1 Plant materials
        3.3.2 Construction of pBINPLUS-CDS and transformation of Agrobacterium tumefaciens strain AGL-0
        3.3.3 Genetic transformation of T cinerariifolium
        3.3.4 Selection and regeneration of transformants
        3.3.5 DNA isolation and PCR analysis
        3.3.6 Phenotype analysis of transgenic T cinerariifolium plants
        3.3.7 RNA isolation and real-time qPCR analysis
        3.3.8 Pyrethrins Extraction methods for HPLC Analysis
        3.3.9 Pyrethrins evaluation by High-performance liquid chromatography （HPLC）
        3.3.10 Statistical analysis
    3.4 Results
    3.5 Discussion
4 Molecular evolution and phylogenetic utility of the chrysanthemyl diphosphatesynthase （TcCDS） gene,resolving relationship within Chrysanthemum
    4.1 Abstrace
    4.2 Introduction
    4.3 Materials and methods
        4.3.1 Plant material and genomic DNA isolation
        4.3.2 Nested PCR amplification and purification
        4.3.3 Cloning and sequencing
        4.3.4 DNA sequenco analysis
    4 Result
        4.1 Gene characterization and genetic analysis
        4.2 Number of nueleotide substitutions per site
        4.3 Synonymous and nonsynonymous substitutions
        4.4 Phylogenetic analysis
    4.5 Discussion
5 General Conclusion and Prospects
    5.1 Molecular Cloning and Characterization of the Trichome Speoitic Chrysanthemyl Diphosphate Promoter from Tanacetum cinerariifolium
    5.2 Overexpression of the genes Chrysanthemyl Diphosphate Synthase（TcCDS）increasedpyrethrins content in Tanacetum cinerariifolium
    5.3 The molecular evolution and phylogenetic utility of the chrysanthemyl diphosphatesynthase （CDS） gene in Chrysanthemum
References
Acknowledgements
Resume
List of Publications


【參考文獻(xiàn)】：
期刊論文
[1]KaKsCalculator:Calculating Ka and Ks Through Model Selection and Model Averaging[J]. Gane Ka-Shu Wong.  Genomics Proteomics & Bioinformatics. 2006(04)
[2]菊花起源的RAPD分析[J]. 戴思蘭,陳俊愉,李文彬.  植物學(xué)報(bào). 1998(11)



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