植物是人類食物的最終來源。為了得到更高產(chǎn)量和更好的品質,人們深入研究了植物多種重要的性狀,包括對生物/非生物脅迫的抗性、產(chǎn)量組成、大量及微量營養(yǎng)和儲存期,單個性狀或多種形狀的組合情況。通常情況下,最成功的作法基于對控制性狀的生物學過程的關鍵酶和/或轉錄因子的操作。盡管人們?yōu)榻沂具@些生物學過程已經(jīng)取得了重要的進展,但是與這些過程明顯無關的性狀是如何協(xié)調調節(jié)的卻仍然知之甚少。本研究中我們鑒定了番茄中的一個類MIXTA的MYB家族基因SIMX1。該基因可以同時調節(jié)能夠決定以上性狀的發(fā)育、生理和代謝過程,有可能正是通過對這些過程中的關鍵酶的轉錄調節(jié)而實現(xiàn)的。過表達SIMX1基因可以增強番茄對廣譜的生物學/非生物學脅迫的抗性。該研究為作物育種領域提供了一個通過操作單個調節(jié)基因而得到多個理想性狀的實例。

【文章頁數(shù)】：103 頁

【學位級別】：博士

【文章目錄】：
ABSTRACT
中文摘要
ABBREVATIONS
1 REVIEW OF LITERATURE
    1.1 Brief History of Tomato
    1.2 Abiotic Stress
        1.2.1 Drought
        1.2.2 Salinity
    1.3 Biotic Stresses
    1.4 Tomato Structure
        1.4.1 Trichome Formation
        1.4.2 Cutin and Wax Biosynthesis
    1.5 Tomato secondary metabolism
        1.5.1 Terpenoids
        1.5.2 Flavonoids
        1.5.3 Carotenoids
    1.6 RNA Sequencing
    1.7 MYB and MIXTA genes
2 MATERIALS and METHODS
    2.1 Plant Material
    2.2 Bioinformatics analysis
    2.3 Vectors construction and plant transformation and regeneration
    2.4 Molecular analysis of transgenic plants
        2.4.1 DNA Extraction
        2.4.2 Transgenic Analysis
        2.4.3 Transcript Test
        2.4.4 Expression Analyses
    2.5 Bimolecular Assays
        2.5.1 Yeast One-Hybrid Assay
        2.5.2 Yeast Two- Hybrid Assay
        2.5.3 Bimolecular Fluorescence Complementation Assay
    2.6 RNA Seq and Bioinformatics analysis
    2.7 Electron, confocal and Light Microscopy
    2.8 Abiotic and biotic Stress Tolerance Assays
        2.8.1 Analysis of Drought and Salt Tolerance
        2.8.2 Stomatal conductance and aperture measurement
        2.8.3 Biotic Stress Tolerance Assays
    2.9 Extraction and Measurement of Secondary Metabolites
        2.9.1 Extraction and Measurement of Terpenoids
        2.9.2 Extraction and Measurement of Flavonoids
        2.9.3 Extraction and measurement of carotenoids
    2.10 Fruit Quality measurements
        2.10.1 Ethylene measurement
        2.10.2 Fruit Firmness
        2.10.3 Shelf Life
        2.10.4 Yield and Yield Component
3 RESULTS
    3.1 Characterization of Sl MX1
    3.2. Cloning of Sl MX1, Transgenic Analysis and Transcript Test
    3.3 Expression profile of Sl MX1 in different organs
    3.4 Sub-cellular Localization of Sl MX1 Protein
    3.5 Phenotypic characterization of Sl MX1 transgenic lines
        3.5.1 Sl MX1 positively regulates trichome formation in tomato
        3.5.2 Sl MX1 positively regulates fruit color in tomato
        3.5.3 Sl MX1 positively regulates cuticle formation and cutin deposition in tomato
        3.5.4 Sl MX1 positively regulates fruit size and tomato yield
    3.6 Sl MX1 positively regulates drought and salt resistant
    3.7 Sl MX1 positively regulates resistant to biotic stresses
        3.7.1 Sl MX1 enhanced resistant to insects
        3.7.2 Sl MX1 enhanced resistant to bacteria
        3.7.3 Sl MX1 enhanced resistant to fungi
        3.7.4 Sl MX1 enhanced resistant to virus
    3.8 Sl MX1 enhanced fruit quality and shelf life
    3.9 Sl MX1 positively enhanced secondary metabolism in tomato
        3.9.1 Sl MX1 positively regulates terpenoid synthesis in tomato
        3.9.2 Sl MX1 positively regulates flavonoid synthesis in tomato
        3.9.3 Sl MX1 positively regulates carotenoid synthesis in tomato
    3.10 RNA Sequencing indicated the positive roles of Sl MX1 in tomato development and metabolism
    3.11 Sl MXl affects the expression of regulatory genes in trichome and cuticle formation
    3.12 Sl MXl affects the expression of key genes in abiotic stress resistance
    3.13 Sl MXl affects the expression of key genes in biotic stress resistance
    3.14 Sl MXl affects the expression of TPS9 a key gene in terpenoid biosynthesis
    3.15 Sl MXl affects the expression of key genes in carotenoid biosynthesis
    3.16 Interaction between Sl MX1 and target genes
    3.17 Interaction between Sl MX1 and Wo
4 DISCUSSION
    4.1 Sl MX1 is a MYB family gene that regulates cuticle development
    4.2 Sl MX1 putatively target Sl Cyc B2 that is crucial for tomato trichome formation
    4.3 Sl MX1 may affect tomato fruit surface formation by regulating the Sl SHNs genes
    4.4 Sl MX1 positively regulates drought and salt tolerance
    4.5 Over expressing Sl MX1 enhanced biotic stress resistance
    4.6 Sl MX1 positively prolonged fruit shelf life and improve tomato yield
    4.7 Over-expression of Sl MX1 is a potentially useful strategy to increase the fruit yield, quality and stress resistance
    4.8 Conclusion
    4.9 Future Perspective
5 REFERENCES
Brief history of Author
Publication
ACKNOWLEDGEMENT



本文編號：3916296