小麥?zhǔn)侨祟惖闹饕澄飦?lái)源之一,中國(guó)是最大的小麥生產(chǎn)國(guó)。確保小麥的優(yōu)質(zhì)和產(chǎn)量,對(duì)人類的生存和發(fā)展和維護(hù)世界和平具有重要意義。然而,小麥生產(chǎn)受到許多因素的影響,尤其是病蟲(chóng)害。由禾谷鐮刀菌（Fusarium graminearum,Fg）引起的小麥赤霉�。‵usarim head blight,FHB）,是世界范圍內(nèi)的一種小麥真菌病害,特別是在中國(guó),造成了巨大的經(jīng)濟(jì)損失。在禾谷鐮刀菌感染植物的過(guò)程中,能夠產(chǎn)生的多種真菌毒素,這些在真菌毒素主要?dú)埩粼谛←湻N子中。真菌毒素嚴(yán)重影響小麥的品質(zhì),在攝入百萬(wàn)分之幾或百萬(wàn)萬(wàn)分之幾微量攝入,對(duì)人畜健康構(gòu)成危險(xiǎn)。小麥赤霉病常發(fā)生在溫暖潮濕的地區(qū),近年來(lái)由于全球變暖,FHB呈現(xiàn)流行病害趨勢(shì)。自2000年以來(lái),在中國(guó)的小麥赤霉病經(jīng)常達(dá)到流行病水平。在過(guò)去幾年中,中國(guó)幾個(gè)小麥主產(chǎn)區(qū)頻繁的爆發(fā)小麥赤霉病的疫情,這越來(lái)越引起人們對(duì)FHB控制的擔(dān)憂。在控制赤霉病的手段中,應(yīng)用殺菌劑成本高昂,而且對(duì)環(huán)境不友好。選育抗病品種是迄今為止控制真菌病害的最佳技術(shù)�？共∮N常用的方法包括常規(guī)選擇、標(biāo)記輔助選擇（Marker-assisted selection,MAS）和基因工程。... 

【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)】：133 頁(yè)

【學(xué)位級(jí)別】：博士

【文章目錄】：
ABSTRACT
摘要
TABLE OF ABBREVIATION
1 GENERAL INTRODUCTION AND LITERATURE REVIEW
    1.1 Overview of Wheat
    1.2 The Fungus Fusarium graminearum
        1.2.1 Taxonomy,Pathology,and Ecology
        1.2.2 Parasitic Life Cycle
        1.2.3 Formation of Mycotoxins
        1.2.4 Symptoms of F.graminearum on Cereals
    1.3 Management of FHB and RNAi
        1.3.1 RNAi Application in Plants against Fungi
        1.3.2 Basic Mechanism of RNAi
        1.3.3 Components of RNAi Machinery
        1.3.4 Different Methods to Trigger RNAi
            1.3.4.1 Host-induced gene silencing（HIGS）
            1.3.4.2 Hairpin RNAi（hp RNAi）
            1.3.4.3 Virus-induced gene silencing（VIGS）
            1.3.4.4 mi RNA induced gene silencing（MIGS）
            1.3.4.5 Using artificial mi RNAs（ami RNAs）
            1.3.4.6 VIGS using ami RNAs（MIR-VIGS）
            1.3.4.7 Spray induced gene silencing（SIGS）
            1.3.4.8DNAi
        1.3.5 Critical Factors for Successful RNAi Induction
    1.4 Protein Kinase C（Pk C）
    1.5 Chitin Synthase7 （Chs7）
    1.6 Aim of this Study
2 MATERIALS AND METHODS
    2.1 Experimental Materials
        2.1.1 Plant Material
        2.1.2 Fungal Strain
        2.1.3 Enzymes,Reagents,and Kits
        2.1.4 Reagents and Culture Media
            2.1.4.1 DNA extraction reagents by CTAB method
            2.1.4.2 Northern blot solutions
    2.2 Test Methods
        2.2.1 Sequence Analysis and3D Structure Prediction
        2.2.2 RNA Extraction by TRIzol
            2.2.2.1 Preparation of samples
            2.2.2.2 RNA extraction
        2.2.3 First Strand c DNA Synthesis
        2.2.4 PCR Amplification（Easy Taq Enzyme）
        2.2.5 Wheat Genomic DNA Extraction by CTAB Method
        2.2.6 Fusarium Head Blight Assay
            2.2.6.1 Preparation of fungal spore suspension
            2.2.6.2 Counting of spores by hemocytometers
            2.2.6.3 Fusarium Seedling blight Assay
            2.2.6.4 Single flower inoculation
        2.2.7 Powdery Mildew Assay
        2.2.8 Northern Blot
            2.2.8.1 Sample Preparation
            2.2.8.2 Gel preparation and electrophoresis
            2.2.8.3 Transfer
            2.2.8.4 Prehybridization
            2.2.8.5 Hybridization
            2.2.8.6 Washing and exposing
        2.2.9 Quantitative RT-PCR Protocol（q RT-PCR）
            2.2.9.1 Primer design criteria
            2.2.9.2 Method
        2.2.10 Mycotoxin Determination by Gass Chromatography-Mass Spectrometry
        2.2.11 Evaluation of Agronomic Traits
        2.2.12 Statistical Analysis
3 RESULTS
    3.1 Identification of Essential Genes
    3.2 Prediction of Protein Structures
    3.3 Prediction of Conserved Domain
    3.4 Multiple Sequence Alignment
    3.5 Construction of Phylogenetic Trees
    3.6 Identification of Gene-Specific Sequence Tags（GSTs）
    3.7 Prediction of Off-Targets
    3.8 Generation of Transgenic Lines
        3.8.1 Transgenic lines developed via biolistic transformation
        3.8.2 Transgenic lines developed via Agrobacterium mediated transformation
    3.9 Establishing Homozygosity and Screening of Transgenic Lines
    3.10 Evaluation of Transgenic Lines against Fusarium Seedling Blight
    3.11 Evaluation of Transgenic Lines against Fusarium Head Blight
    3.12 Quantification of Fungal Transcripts Levels in Transgenic Lines
    3.13 Detection of si RNAs in Transgenic Lines
    3.14 Evaluation of Mycotoxin Resistance in Transgenic Lines
    3.15 Evaluation of Transgenic Lines against Powdery Mildew Disease
    3.16 Evaluation of Agronomic Traits in Transgenic Lines
4 DISCUSSION
    4.1 Selection of Essential Genes
    4.2 Sequence Analysis of Genes
    4.3 Factors Affecting RNAi Efficiency
    4.4 Factors Affecting Transformation Efficiency
    4.5 Evaluation of Transgenic Lines
    4.6 Future Prospects
REFERENCES
Acknowledgment



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