小麥?zhǔn)鞘澜缟系诙蠹Z食作物,已報(bào)道有50多種病毒侵染小麥從而引起嚴(yán)重的產(chǎn)量損失。本研究首先調(diào)查了近年來以大麥黃矮病毒（Barley yellow dwarf viruses）為代表的小麥重要病毒在中國小麥主產(chǎn)區(qū)分布情況;其次是比較了中國和捷克兩國大麥黃矮病毒BYDV-PAV的外殼蛋白（Coat Protein,CP）和通讀蛋白（Readthrough protein,RTP）基因序列的分子變異。此外,還鑒定出WDV傳毒介體異沙葉蟬體內(nèi)存在一種新的軟腐病毒。2017-2019年共采集到來自中國不同地區(qū)小麥、大麥和燕麥的747株疑似病毒病癥狀的樣品。采用多重PCR法同時(shí)檢測了待測樣品中是否存在大麥黃矮病毒（BYDV-PAV,BYDV-GAV,和BYDV-GPV）,小麥矮縮病毒（Wheat dwarf virus,WDV）,大麥黃條點(diǎn)花葉病毒（Barley yellow striate mosaic virus,BYSMV）,水稻黑條矮縮病毒（Rice black-streaked dwarf virus,RBSDV）和北方禾谷花葉病毒（Northern cereal mosaic viru... 

【文章來源】：中國農(nóng)業(yè)科學(xué)院北京市

【文章頁數(shù)】：134 頁

【學(xué)位級別】：博士

【文章目錄】：
中文摘要
Abstract
Abbreviations
CHAPTER-Ⅰ Literature review
    1.1 Wheat production in China
    1.2 Wheat viruses
        1.2.1 Barley yellow dwarf viruses(B/CYDVs)
            1.2.1.1 BYDV-PAV
            1.2.1.2 BYDV-GAV
            1.2.1.3 BYDV-GPV
        1.2.2 Wheat dwarf virus(WDV)
        1.2.3 Barley yellow striate mosaic virus(BYSMV)
        1.2.4 Northern cereal mosaic virus(NCMV)
        1.2.5 Rice black streaked dwarf virus(RBSDV)
    1.3 Plant virus transmission
        1.3.1 Aphid transmitted virus
        1.3.2 Leafhopper and planthopper transmitted virus
    1.4 Molecular approaches to virus detection
        1.4.1 Reverse transcriptase polymerase chain reaction(RT-PCR)
        1.4.2 Multiplex RT-PCR
    1.5 Molecular evolution of RNA viruses
    1.6 Sanger sequencing
    1.7 Next generation sequencing
    1.8 Rapid amplification c DNA ends(RACE)
    1.9 Objectives
CHAPTER-Ⅱ Occurrence of wheat viruses in different regions of China during 2017,2018 and 2019
    2.1 Introduction
    2.2 Materials and methods
        2.2.1 Samples collection
        2.2.2 Total RNA extraction
        2.2.3 Detection of wheat viruses by using Multiplex RT-PCR
            2.2.3.1 Multiplex PCR procedure for BYDV-PAV, -GAV and–GPV
            2.2.3.2 Multiplex PCR procedure for WDV,BYSMV,NCMV and RBSDV
    2.3 Results
        2.3.1 Viruses prevalence
        2.3.2 Virus co-infection
    2.4 Discussion
CHAPTER-Ⅲ Comparison of BYDV-PAV CP and RTP genes among different regions of China
    3.1 Introduction
    3.2 Materials and methods
        3.2.1 Samples collection
        3.2.2 Detection of BYDV-PAV using Multiplex RT-PCR
        3.2.3 Detection of BYDV-PAV CP and RTP
        3.2.4 Purification of PCR products
        3.2.5 Ligation to pEASY-T5 vector
        3.2.6 Transformation of pEASY-T5 vector ligation product into E.coli
        3.2.7 Sequence alignment and phylogenetic analysis
    3.3 Results
    3.4 Discussion
CHAPTER-Ⅳ Molecular diversity of BYDV-PAV CP and RTP from China and the Czech Republic
    4.1 Introduction
    4.2 Materials and methods
        4.2.1 Samples collection
        4.2.2 RNA extraction
        4.2.3 RT-PCR
            4.2.3.1 Detection of BYDV-PAV
            4.2.3.2 Detection of BYDV-PAV CP and RTP genes
        4.2.4 Purification of PCR products
        4.2.5 Ligation to pEASY-T5 vector
        4.2.6 Transformation of pEASY-T5 vector ligation product into E.coli
        4.2.7 Sequence alignments
        4.2.8 Phylogenetic analysis and population diversity
    4.3 Results
        4.3.1 Nucleotide identity position and phylogenetic analysis
        4.3.2 Population diversity and neutrality
    4.4 Discussion
CHAPTER-Ⅴ The complete genomic sequence of a novel iflavirus in leafhopper
    5.1 Introduction
    5.2 Materials and methods
        5.2.1 Leafhopper collection,viral metagenomic and bioinformatics analysis
        5.2.2 Total RNA extraction
        5.2.3 Sequencing of the viral genome and sequence analysis
        5.2.4 Rapid amplification of c DNA ends(RACE)
            5.2.4.1 3'RACE
            5.2.4.2 5'RACE
        5.2.5 Purification,ligation and transformation Procedure
        5.2.6 Analysis of the nucleotide sequence and evolutionary relationship
    5.3 Results
        5.3.1 Open reading frame analysis and genome organization
        5.3.2 Structural and non-structural protein
        5.3.3 Phylogenetic relationship to other viruses in order Picornavirales and family Iflavirus
    5.4 Discussion
CHAPTER-Ⅵ Conclusion
References
Acknowledgements
Author biography
Appendix


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