大豆胞囊线虫4号小种抗性候选基因GmSNAP11的克隆与功能验证
发布时间：2021-10-16 00:02
大豆胞囊线虫病（SCN,Heterodera glycines Ichinohe）是世界性的重要病害,每年可导致几十亿美元的经济损失。在大豆胞囊线虫病的防控策略中,培育抗性品种是行之最为有效的方法。鉴于大豆胞囊线虫病的危害严重性,开展大豆SCN抗性基因/位点的发掘具有重要意义。目前,由于混合群体分离法的新一代测序技术（BSA-seq）可以显著加速豆类作物的抗病基因的鉴定和发掘效率,因此采用该技术来快速鉴定大豆胞囊线虫病的抗性基因/位点是可行的。本研究主要针对大豆胞囊线虫病4号小种（SCN4）采用BSA-seq方法发现了一个新的抗性基因位点和相应候选基因,初步验证了基因功能,并开发出了诊断分子标记,可应用于大豆胞囊线虫病抗性育种中。本研究首先利用晋豆23（敏感品种）×灰皮支黑豆（抗性品种）衍生的重组自交系群体145个家系为材料,采用胞囊指数的方法鉴定该群体对SCN4号小种的抗病指数,然后依据鉴定结果构建了各包含15个家系的抗病和感病基因池,基于BSA-seq的方法分别对抗病与感基因池以及双亲进行重测序,依据欧式距离（ED）和SNP指数（SNP-index）的方法分析比较抗感池和双亲之间的...
【文章来源】：中国农业科学院北京市
【文章页数】：109 页
【学位级别】：博士
【文章目录】：
附件
摘要
Abstract
List of abbreviations
Chapter1 Background Information and Literature Review
1.1 Introduction
1.2 The life cycle of soybean cyst nematode
1.3 SCN Population and race
1.4 Distribution of SCN
1.5 Management of SCN in Soybean
1.6 Cytological and histological process of syncytium
1.7 Germplasm screening and sources of SCN resistance
1.8 Identification of SCN resistant QTL and genes
1.9 Functional gene analysis studies for SCN resistance
1.10 Molecular mechanisms of SCN resistance
1.11 Soybean Breeding Strategies for Resistance to SCN
1.12 Summary
1.13 Objectives
1.14 Technical route map
Chapter2 Candidate Genes Mining for Resistance to Soybean Cyst Nematode Race4 Based on Bulk Segregant Analysis in Glycine max
2.1 Introduction
2.2 Materials and methods
2.2.1 Plant materials and phenotypic analysis
2.2.2 Construction of Sequencing Libraries and Sequencing
2.2.3 Identification of genomic regions and putative candidate genes for SCN
2.3 Results
2.3.1 Phenotypic variations and construction of extreme bulks for SCN4 resistance
2.3.2 Whole-genome resequencing,mapping of reads and identification of SNPs
2.3.3 Identification of candidate genomic regions for SCN4 resistance
2.3.4 Putative candidate genes for SCN4 resistance
2.3.5 No copy number variation was observed in rhg1-paralog
2.4 Discussion
2.5 Summary
Chapter3 Functional Analysis of Gm SNAP11 for Resistance to Soybean Cyst Nematode Race4 in Glycine max
3.1 Introduction
3.2 Materials and methods
3.2.1 Plant materials and phenotypic analysis
3.2.2 Sequencing and identification of genetic variation
3.2.3 Functional analysis of Gm SNAP11 and Gm PLAC8
3.2.4 Hairy transformation system
3.2.5 Nematode Infection of Transgenic Hairy Roots
3.2.6 RNA Isolation and q PCR
3.3 Results
3.3.1 Sequencing and identification of genetic variation of Gm SNAP11
3.3.2 Gm SNAP11 contributes to SCN4 resistance
3.4 Discussion
3.5 Summary
Chapter4 Development and Utilization of KASP Markers for the Soybean Cyst Nematode Resistance loci,rhg1,Rhg4 and rhg1-paralog
4.1 Introduction
4.2 Materials and methods
4.2.1 Plant materials and phenotyping
4.2.2 SNP identification and development of Kompetitive allele-specific PCR(KASP)genotyping assays
4.2.3 SNP marker development and validation of candidate genomic regions identified
4.2.4 Statistical analysis
4.3 Results
4.3.1 Application of KASP assays for selection of resistance to SCN
4.3.2 The presence of three resistant loci exhibit a higher effect for SCN4 resistance
4.4 Discussion
4.5 Summary
Chapter5 Conclusions
References
Appendix
Acknowledgements
Resume
【参考文献】：
期刊论文
[1]Deep genotyping of the gene GmSNAP facilitates pyramiding resistance to cyst nematode in soybean[J]. Yu Tian,Bo Liu,Xuehui Shi,Jochen C.Reif,Rongxia Guan,Ying-hui Li,Li-juan Qiu. The Crop Journal. 2019(05)
[2]抗大豆胞囊线虫SCN3-11位点的KASP标记开发和利用[J]. 田宇,杨蕾,李英慧,邱丽娟. 作物学报. 2018(11)
[3]Soybean hairy roots produced in vitro by Agrobacterium rhizogenes-mediated transformation[J]. Li Chen,Yupeng Cai,Xiujie Liu,Chen Guo,Shi Sun,Cunxiang Wu,Bingjun Jiang,Tianfu Han,Wensheng Hou. The Crop Journal. 2018(02)
[4]大豆胞囊线虫主效抗病基因Rhg4（GmSHMT）的CAPS/dCAPS标记开发和利用[J]. 史学晖,李英慧,于佰双,郭勇,王家军,邱丽娟. 作物学报. 2015(10)
[5]中国大豆抗(耐)胞囊线虫病品种及其系谱分析[J]. 袁翠平,沈波,董英山. 大豆科学. 2009(06)
[6]基于大豆胞囊线虫病抗性候选基因rhg1的InDel标记开发与鉴定[J]. 南海洋,李英慧,常汝镇,邱丽娟. 作物学报. 2009(07)
[7]大豆胞囊线虫病抗源筛选及应用研究进展[J]. 刘佩印. 黑龙江农业科学. 2005(06)
[8]我国大豆孢囊线虫抗源筛选及抗病育种研究进展[J]. 崔文馥. 大豆科学. 1998(01)
[9]大豆根渗出物对大豆孢囊线虫4号生理小种卵孵化的影响[J]. 颜清上,陈品三,王连铮. 植物病理学报. 1997(03)
[10]中国小黑豆抗源对大豆孢囊线虫4号生理小种抗性机制的研究Ⅱ.抗感品种根部合胞体超微结构的比较[J]. 颜清上,陈品三,王连铮. 植物病理学报. 1997(01)
本文编号：3438843
【文章来源】：中国农业科学院北京市
【文章页数】：109 页
【学位级别】：博士
【文章目录】：
附件
摘要
Abstract
List of abbreviations
Chapter1 Background Information and Literature Review
1.1 Introduction
1.2 The life cycle of soybean cyst nematode
1.3 SCN Population and race
1.4 Distribution of SCN
1.5 Management of SCN in Soybean
1.6 Cytological and histological process of syncytium
1.7 Germplasm screening and sources of SCN resistance
1.8 Identification of SCN resistant QTL and genes
1.9 Functional gene analysis studies for SCN resistance
1.10 Molecular mechanisms of SCN resistance
1.11 Soybean Breeding Strategies for Resistance to SCN
1.12 Summary
1.13 Objectives
1.14 Technical route map
Chapter2 Candidate Genes Mining for Resistance to Soybean Cyst Nematode Race4 Based on Bulk Segregant Analysis in Glycine max
2.1 Introduction
2.2 Materials and methods
2.2.1 Plant materials and phenotypic analysis
2.2.2 Construction of Sequencing Libraries and Sequencing
2.2.3 Identification of genomic regions and putative candidate genes for SCN
2.3 Results
2.3.1 Phenotypic variations and construction of extreme bulks for SCN4 resistance
2.3.2 Whole-genome resequencing,mapping of reads and identification of SNPs
2.3.3 Identification of candidate genomic regions for SCN4 resistance
2.3.4 Putative candidate genes for SCN4 resistance
2.3.5 No copy number variation was observed in rhg1-paralog
2.4 Discussion
2.5 Summary
Chapter3 Functional Analysis of Gm SNAP11 for Resistance to Soybean Cyst Nematode Race4 in Glycine max
3.1 Introduction
3.2 Materials and methods
3.2.1 Plant materials and phenotypic analysis
3.2.2 Sequencing and identification of genetic variation
3.2.3 Functional analysis of Gm SNAP11 and Gm PLAC8
3.2.4 Hairy transformation system
3.2.5 Nematode Infection of Transgenic Hairy Roots
3.2.6 RNA Isolation and q PCR
3.3 Results
3.3.1 Sequencing and identification of genetic variation of Gm SNAP11
3.3.2 Gm SNAP11 contributes to SCN4 resistance
3.4 Discussion
3.5 Summary
Chapter4 Development and Utilization of KASP Markers for the Soybean Cyst Nematode Resistance loci,rhg1,Rhg4 and rhg1-paralog
4.1 Introduction
4.2 Materials and methods
4.2.1 Plant materials and phenotyping
4.2.2 SNP identification and development of Kompetitive allele-specific PCR(KASP)genotyping assays
4.2.3 SNP marker development and validation of candidate genomic regions identified
4.2.4 Statistical analysis
4.3 Results
4.3.1 Application of KASP assays for selection of resistance to SCN
4.3.2 The presence of three resistant loci exhibit a higher effect for SCN4 resistance
4.4 Discussion
4.5 Summary
Chapter5 Conclusions
References
Appendix
Acknowledgements
Resume
【参考文献】：
期刊论文
[1]Deep genotyping of the gene GmSNAP facilitates pyramiding resistance to cyst nematode in soybean[J]. Yu Tian,Bo Liu,Xuehui Shi,Jochen C.Reif,Rongxia Guan,Ying-hui Li,Li-juan Qiu. The Crop Journal. 2019(05)
[2]抗大豆胞囊线虫SCN3-11位点的KASP标记开发和利用[J]. 田宇,杨蕾,李英慧,邱丽娟. 作物学报. 2018(11)
[3]Soybean hairy roots produced in vitro by Agrobacterium rhizogenes-mediated transformation[J]. Li Chen,Yupeng Cai,Xiujie Liu,Chen Guo,Shi Sun,Cunxiang Wu,Bingjun Jiang,Tianfu Han,Wensheng Hou. The Crop Journal. 2018(02)
[4]大豆胞囊线虫主效抗病基因Rhg4（GmSHMT）的CAPS/dCAPS标记开发和利用[J]. 史学晖,李英慧,于佰双,郭勇,王家军,邱丽娟. 作物学报. 2015(10)
[5]中国大豆抗(耐)胞囊线虫病品种及其系谱分析[J]. 袁翠平,沈波,董英山. 大豆科学. 2009(06)
[6]基于大豆胞囊线虫病抗性候选基因rhg1的InDel标记开发与鉴定[J]. 南海洋,李英慧,常汝镇,邱丽娟. 作物学报. 2009(07)
[7]大豆胞囊线虫病抗源筛选及应用研究进展[J]. 刘佩印. 黑龙江农业科学. 2005(06)
[8]我国大豆孢囊线虫抗源筛选及抗病育种研究进展[J]. 崔文馥. 大豆科学. 1998(01)
[9]大豆根渗出物对大豆孢囊线虫4号生理小种卵孵化的影响[J]. 颜清上,陈品三,王连铮. 植物病理学报. 1997(03)
[10]中国小黑豆抗源对大豆孢囊线虫4号生理小种抗性机制的研究Ⅱ.抗感品种根部合胞体超微结构的比较[J]. 颜清上,陈品三,王连铮. 植物病理学报. 1997(01)
本文编号：3438843
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