水稻是世界上重要的糧食作物之一,也是中國的主要糧食作物。稻米養(yǎng)活了世界一半以上的人口,中國近65%的人口是以稻米為食,培育出產(chǎn)量更高更穩(wěn)的水稻新品種有利于保證國家的糧食安全。近年來,水稻基因組學(xué)的發(fā)展給水稻優(yōu)良品種的選育帶來了新的機(jī)遇和挑戰(zhàn)。正常的葉色有利于葉片進(jìn)行光合作用,而一些特殊的葉色也可作為鑒定水稻材料的有效標(biāo)志性狀。葉色的變化與植株衰老有著直接或間接的關(guān)系,對水稻的產(chǎn)量和品質(zhì)影響很大。目前,在水稻中已經(jīng)發(fā)現(xiàn)了一些與葉色有關(guān)的基因,但是對于調(diào)控葉色與葉片衰老的遺傳機(jī)理和分子機(jī)制仍缺乏充分的了解和認(rèn)識。深入研究這些葉色基因的生物學(xué)功能和代謝途徑有助于葉色標(biāo)記性狀在水稻遺傳育種中的應(yīng)用,在水稻抗衰老育種中也有重要的理論和實(shí)踐意義。本研究采用甲基磺酸乙酯（EMS）誘變技術(shù)從秈稻品種浙農(nóng)34的M2群體中分離到兩個不同的葉色突變體,分別命名為水稻葉片主脈褐色突變體bml和紫葉色突變體pl。并采用表型觀察、圖位克隆和基因表達(dá)分析等方法分析了突變基因的功能。結(jié)果表明,兩個葉色基因控制的表型能夠穩(wěn)定遺傳,在葉片衰老過程中均起著重要作用,其中OsBML和OsPL分別參與了早期和后期的葉片衰老調(diào)控... 

【文章來源】：浙江大學(xué)浙江省 211工程院校 985工程院校 教育部直屬院校

【文章頁數(shù)】：107 頁

【學(xué)位級別】：博士

【文章目錄】：
Acknowledgements
List of Abbreviations
Abstract
摘要
1 Introduction
2 Literature review
    2.1 Research progress of color mutants
    2.2 Source of leaf color mutants
    2.3 Types and physiological changes of leaf color mutants
        2.3.1 Types of leaf color mutants
        2.3.2 Physiological changes in leaf color mutant
    2.4 Factors affecting rice leaf coloring
        2.4.1 Environmental factors affecting leaf senescence in rice
        2.4.2 Cultivation conditions affecting leaf senescence in rice
        2.4.3 Plant factors affecting the leaf coloring of rice leaves
    2.5 Molecular mechanism of rice leaf color
        2.5.1 Chloroplast development related genes
        2.5.2 Chlorophyll metabolism related genes
        2.5.3 Gene mutations in the heme metabolism pathway
        2.5.4 Gene mutations encoding other chloroplast proteins
        2.5.5 Gene mutation not directly related to the photosynthetic system
    2.6 Leaf senescence and its regulation of coloration in rice
        2.6.1 Genetic mechanism of rice leaf senescence
        2.6.2 Chloroplast development and chlorophyll metabolism related genes
        2.6.3 Transcription factor gene
            2.6.3.1 Classification of MYB transcription factor genes
            2.6.3.2 Metabolic path way for MYB transcription factor
            2.6.3.3 Advances in the regulation of anthocyanin synthesis
        2.6.4 Hormone metabolism related genes
        2.6.5 Protein modification related genes
            2.6.5.1 Analysis and comparison of RLCK and RLK characteristics
            2.6.5.2 Functional analysis of RLCK
        2.6.6 Other aging related genes
    2.7 The genetic analysis for the leaf color and leaf senescence
    2.8 Purpose and significance of present study
3 Materials and methods
    3.1 Rice materials and growth conditions
    3.2 Agronomic traits measurement
    3.3 Gene mapping program
        3.3.1 DNA extraction method
        3.3.2 SSR and InDel molecular marker analysis method
    3.4 Gene mapping
    3.5 Phylogenic analysis
    3.6 Different physiological parameters analysis
        3.6.1 Estimation of the chlorophyll content, photosynthetic and chlorophyllfluorescence parameters
        3.6.2 Anthocyanin content measurement
        3.6.3 Microscopic structure analysis
        3.6.4 Transmission electron microscopy （TEM）
        3.6.5 Histochemistry assay
        3.6.6 Measurement of antioxidant and enzyme activity
        3.6.7 Measurement of endogenous phytohormones
        3.6.8 Dark-induced treatment
        3.6.9 Inoculation with bacterial blight pathogen
        3.6.10 Temperature treatment on pl
    3.7 Total RNA extraction
    3.8 Quantitative qRT-PCR analysis
    3.9 Construction of artificial RNA interference vector of OsBML
    3.10 Genetically modified tissue culture procedures in rice
        3.10.1 Callus induction and subculture
        3.10.2 Infection of rice callus
        3.10.3 Co-cultivation
        3.10.4 Selection of resistant callus
        3.10.5 Pre-distribution
        3.10.6 Differentiation
        3.10.7 Rooting
    3.11 Statistical Analysis
4 Results and analysis
    4.1 Phenotypic analysis of mutants
        4.1.1 The analysis for the bml mutants
            4.1.1.1 Phenotypic trait performance of the bml mutants
            4.1.1.2 Agronomic and yield related traits characterization of the bml mutant394.1.1.3 Chlorophyll content, photosynthesis and chlorophyll fluorescence ofthe bml mutant
            4.1.1.4 Ultrastructure changes in mesophyll cells and leaf chloroplasts of bmlmutant
            4.1.1.5 Activities of antioxidant enzymes and reactive oxygen species in thebml mutant
            4.1.1.6 Measurements of phytohonnones in the bml mutant
        4.1.2 The analysis for the pl mutant
            4.1.2.1 Phenotypic trait characterization of the pl mutant
            4.1.2.2 Chlorophyll contents, chlorophyll fluorescence in pl mutant
            4.1.2.3 Anthocyanin contents analysis
            4.1.2.4 Ultra structure alterations in leaf mesophyll cells in the pl mutant
            4.1.2.5 Antioxidant enzymes, MDA and TSS contents in the pl mutant
            4.1.2.6 Phytohormones measurement in the pl mutant
    4.2 Genetic analysis and gene mapping of color leaf mutants
        4.2.1 Genetic analysis, map-based cloning and confirmation of the mutation of theOsBML gene
            4.2.1.1 Genetic analysis and map-based cloning of the OsBML gene
            4.2.1.2 Confirmation of the mutation of the OsBML gene
        4.2.2 Genetic analysis and map-based cloning of OsPL gene
    4.3 Phylogenetic analysis and domain location
        4.3.1 Phylogenetic analysis and domain location of OsBML gene
        4.3.2 Phytogenic tree analysis of OsPL gene
    4.4 Expression pattern analysis
        4.4.1 Expression pattern analysis of OsBML gene
        4.4.2 Leaf senescence-associated gene expression in the bml mutant
        4.4.3 Disease response and upregulation of pathogenesis-related marker genes
        4.4.4 Expression pattern analysis of OsPL gene
        4.4.5 Photosynthesis and anthocyanin biosynthesis associated gene expression inthe pl mutant
        4.4.6 Effect of high temperature on pl mutant
5 Discussions
    5.1 Change of normal leaf color leads directly or indirectly to leaf senescence
    5.2 BML gene involved in the induction of defense responses against disease
    5.3 OsPL gene involved in anthocyanin synthesis, leaf senescence and heat stressresponse
6 References
7 List of publications


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