小麦重要病毒的发生动态和中捷两国大麦黄矮病毒PAV的分子差异
发布时间：2021-11-18 07:36
小麦是世界上第二大粮食作物,已报道有50多种病毒侵染小麦从而引起严重的产量损失。本研究首先调查了近年来以大麦黄矮病毒（Barley yellow dwarf viruses）为代表的小麦重要病毒在中国小麦主产区分布情况;其次是比较了中国和捷克两国大麦黄矮病毒BYDV-PAV的外壳蛋白（Coat Protein,CP）和通读蛋白（Readthrough protein,RTP）基因序列的分子变异。此外,还鉴定出WDV传毒介体异沙叶蝉体内存在一种新的软腐病毒。2017-2019年共采集到来自中国不同地区小麦、大麦和燕麦的747株疑似病毒病症状的样品。采用多重PCR法同时检测了待测样品中是否存在大麦黄矮病毒（BYDV-PAV,BYDV-GAV,和BYDV-GPV）,小麦矮缩病毒（Wheat dwarf virus,WDV）,大麦黄条点花叶病毒（Barley yellow striate mosaic virus,BYSMV）,水稻黑条矮缩病毒（Rice black-streaked dwarf virus,RBSDV）和北方禾谷花叶病毒（Northern cereal mosaic viru...
【文章来源】：中国农业科学院北京市
【文章页数】：134 页
【学位级别】：博士
【文章目录】：
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
【参考文献】：
期刊论文
[1]灰飞虱传播的一种小麦病毒病鉴定[J]. 邸垫平,张永亮,张爱红,闫冲,杨菲,路银贵,田兰芝,王献兵,苗洪芹. 植物病理学报. 2016(04)
[2]中国北方四省小麦丛矮病病原鉴定[J]. 段西飞,邸垫平,张爱红,苗洪芹. 植物病理学报. 2013(01)
[3]水稻黑条矮缩病毒p8蛋白的原核表达、抗血清制备及其特性[J]. 羊健,张恒木,陈剑平,戴良英. 植物保护学报. 2007(03)
[4]The complete nucleotide sequence and its organization of the genome of Barley yellow dwarf virus-GAV[J]. JIN Zhibo, WANG Xifeng, CHANG Shengjun & ZHOU Guanghe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sci-ences, Beijing 100094, China. Science in China（Series C:Life Sciences）. 2004(02)
[5]Identification of a protein associated with circulative transmission of Barley yellow dwarf virus from cereal aphids, Schizaphis graminum and Sitobion avenae[J]. WANG Xifeng & ZHOU Guanghe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China. Chinese Science Bulletin. 2003(19)
[6]大麦黄矮病毒PAV中国分离物外壳蛋白基因的克隆及同源性分析[J]. 贾力,吴茂森,张文蔚,成卓敏. 植物病理学报. 2001(03)
[7]一种由麦二叉蚜、禾缢管蚜非专化性传毒的小麦黄矮病毒株系鉴定[J]. 张淑香,周广和. 植物病理学报. 1987(02)
[8]小麦黄矮病毒4种株系鉴定与应用[J]. 周广和,张淑香,钱幼亭. 中国农业科学. 1987(04)
[9]我国禾谷类病毒病的病原问题——Ⅷ.玉米粗缩病病原的研究[J]. 龚祖埙,沈菊英,陈巽祯,刘信义. Acta Biochimica et Biophysica Sinica. 1981(01)
[10]小麦丛矮病研究之一分布、症状及损失[J]. 裘维蕃,杨莉,梅汝鸿,林志亮,蔡祝南. 植物保护学报. 1979(01)
本文编号：3502485
【文章来源】：中国农业科学院北京市
【文章页数】：134 页
【学位级别】：博士
【文章目录】：
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
【参考文献】：
期刊论文
[1]灰飞虱传播的一种小麦病毒病鉴定[J]. 邸垫平,张永亮,张爱红,闫冲,杨菲,路银贵,田兰芝,王献兵,苗洪芹. 植物病理学报. 2016(04)
[2]中国北方四省小麦丛矮病病原鉴定[J]. 段西飞,邸垫平,张爱红,苗洪芹. 植物病理学报. 2013(01)
[3]水稻黑条矮缩病毒p8蛋白的原核表达、抗血清制备及其特性[J]. 羊健,张恒木,陈剑平,戴良英. 植物保护学报. 2007(03)
[4]The complete nucleotide sequence and its organization of the genome of Barley yellow dwarf virus-GAV[J]. JIN Zhibo, WANG Xifeng, CHANG Shengjun & ZHOU Guanghe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sci-ences, Beijing 100094, China. Science in China（Series C:Life Sciences）. 2004(02)
[5]Identification of a protein associated with circulative transmission of Barley yellow dwarf virus from cereal aphids, Schizaphis graminum and Sitobion avenae[J]. WANG Xifeng & ZHOU Guanghe State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China. Chinese Science Bulletin. 2003(19)
[6]大麦黄矮病毒PAV中国分离物外壳蛋白基因的克隆及同源性分析[J]. 贾力,吴茂森,张文蔚,成卓敏. 植物病理学报. 2001(03)
[7]一种由麦二叉蚜、禾缢管蚜非专化性传毒的小麦黄矮病毒株系鉴定[J]. 张淑香,周广和. 植物病理学报. 1987(02)
[8]小麦黄矮病毒4种株系鉴定与应用[J]. 周广和,张淑香,钱幼亭. 中国农业科学. 1987(04)
[9]我国禾谷类病毒病的病原问题——Ⅷ.玉米粗缩病病原的研究[J]. 龚祖埙,沈菊英,陈巽祯,刘信义. Acta Biochimica et Biophysica Sinica. 1981(01)
[10]小麦丛矮病研究之一分布、症状及损失[J]. 裘维蕃,杨莉,梅汝鸿,林志亮,蔡祝南. 植物保护学报. 1979(01)
本文编号：3502485
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