新型棉花R2R3转录因子GaMYB62L和GaMYB85的功能分析
发布时间：2022-06-01 21:22
MYB转录因子是非常著名的一类转录因子,它们主要参与胁迫响应的基因互做和信号通路。MYB转录因子在氨基酸的N端含有一个保守的、具有53个氨基酸残基的DNA结合结构域—MYB结构域,MYB结构域可以折叠成螺旋-环-螺旋结构。近期研究发现MYB结构域也可位于MYB转录因子氨基酸的C端。依据DNA结合结构域的不同,MYB蛋白可以进一步分成四类:MYB-1R,R2R3-MYB,R1R2R3-MYB（MYB3R）和4-RMYB类蛋白。R1R2R3-MYB类蛋白广泛的存在于单子叶植物和双子叶植物中,棉花D基因组是目前为止含有R2R3-MYB基因最多的基因组（相较于拟南芥126个）,大约具有205个。R2R3-MYB基因参与许多胁迫响应,植物发育和细胞内代谢过程。R2R3-MYB转录因子涉及植物对多种非生物胁迫的应答,但是对棉花中R2R3-MYB基因参与干旱胁迫应答的研究还比较少。而且,在干旱胁迫下的分子网络和植物基因的功能仍然不是很清楚。在本研究中,我们克隆了两个来自亚洲棉的R2R3-MYB基因GaMYB62L和GaMYB85,并将它们转化于拟南芥中。这两个基因的过表达拟南芥展现出耐受干旱的表型,...
【文章页数】：114 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER ONE INTRODUCTION
1.1 COTTON (GOSSYPIUM SPP.)
1.2 MYB FAMILY
1.3 EVOLUTION OF MYB DNA BINDING DOMAINS
1.4 MYB TRANSCRIPTION FACTORS AND DROUGHT TOLERANCE
1.5 GENETIC APPROACHES AND MOLECULAR REGULATION FOR DROUGHT TOLERANCE
1.6 HYPOTHESIS
1.7 RESEARCH OBJECTIVES
CHAPTER TWO FUNCTIONAL CHARACTERIZATION OF COTTON GAMYB62L, A NOVEL R2R3 TFIN TRANSGENIC ARABIDOPSIS
2.1 MATERIAL AND METHODS
2.1.1 Bioinformatic analysis of GaMYB62L gene
2.1.2 Plant transformation and screening of 35S: GaMYB62L Arabidopsis lines
2.1.3 Response of overexpressed GaMYB62L transgenic lines to drought and cold stress
2.1.4 Germination and root elongation assays
2.1.5 Measurements of transpirational Water loss rate and Relative water content
2.1.6 Analysis of proline content and chlorophyll contents
2.1.7 Stomatal opening rate determination in response to Abscisic acid treatment
2.1.8 Gene expression analysis of Markers genes by qRT-PCR
2.1.9 Statistical analysis of Data
2.2 RESULTS
2.2.1 GaMYB62L Sequence Characterization and Phylogenetic analysis
2.2.2 GaMYB62L overexpression positively modulates drought tolerance in transgenic linesof Arabidopsis
2.2.3 Analysis of proline and chlorophyll content in GaMYB62L transgenic lines
2.2.4 Constitutive expression of 35S:GaMYB62L in Arabidopsis up-regulates tolerance toABA, salt and osmotic stresses
2.2.5 Reduced leaf Stomatal Density, size and stomatal opening in 35S:GaMYB62L
2.2.6 Cold response of GaMYB62L-overexpressing Arabidopsis plants
2.2.7 Transcripts investigation of abiotic stress-responsive genes
2.3 DISCUSSION
CHAPTER THREE GAMYB85, NOVEL COTTON R2R3 TF TRANSFORMED IN ARABIDOPSIS HASIMPORTANT ROLE IN DROUGHT TOLERANCE
3.1 MATERIAL AND METHODS
3.1.1 Bioinformatic analysis of GaMYB85 gene
3.1.2 Transgenic plant construction and screening
3.1.3 Germination and root elongation assays
3.1.4 Drought and freezing tolerance assay
3.1.5 Measurements of transpirational WLR and RWC
3.1.6 Proline content and chlorophyll content
3.1.7 Stomata measurements and opening rates determination in response to Abscisic acidtreatment
3.1.8 Gene expression analysis of marker genes by qRT-PCR
3.1.9 Statistical analysis of Data
3.2 RESULTS
3.2.1 GaMYB85 characterization and phylogenetic analysis
3.2.2 Overexpression of 35S:GaMYB85 in plants confers ABA hypersensitivity for seedgermination and root elongation
3.2.3 Overexpression of 35S:GaMYB85 in plants confers enhanced salt and osmotictolerance
3.2.4 Enhanced GaMYB85 tolerance under drought and freezing stress
3.2.5 Reduced stomatal density, increased stomatal response, and increased stomata sizein GaMYB85 plants
3.2.6 Abiotic stress responsive marker genes transcript analysis
3.3 DISCUSSION
CHAPTER FOUR CONCLUSION
References
SUPPLEMENTARY DATA I
SUPPLEMENTARY DATA II
ACKNOWLEDGEMENT
RESUME
【参考文献】：
期刊论文
[1]Arabidopsis C3HC4-RING finger E3 ubiquitin ligase At AIRP4 positively regulates stress-responsive abscisic acid signaling[J]. Liang Yang,Qiaohong Liu,Zhibin Liu,Hao Yang,Jianmei Wang,Xufeng Li,Yi Yang. Journal of Integrative Plant Biology. 2016(01)
[2]Transgenic expression of MYB15 confers enhanced sensitivity to abscisic acid and improved drought tolerance in Arabidopsis thaliana[J]. Zhenhua Ding a, b, Shiming Li a, b, Xueli An a, Xin Liu a, Huanju Qin a, Daowen Wang a, a State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China b Graduate School of Chinese Academy of Sciences, Beijing 100039, China. 遗传学报. 2009(01)
[3]利用SSR标记技术研究棉属A、D染色体组的进化[J]. 郭旺珍,王凯,张天真. 遗传学报. 2003(02)
本文编号：3652859
【文章页数】：114 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER ONE INTRODUCTION
1.1 COTTON (GOSSYPIUM SPP.)
1.2 MYB FAMILY
1.3 EVOLUTION OF MYB DNA BINDING DOMAINS
1.4 MYB TRANSCRIPTION FACTORS AND DROUGHT TOLERANCE
1.5 GENETIC APPROACHES AND MOLECULAR REGULATION FOR DROUGHT TOLERANCE
1.6 HYPOTHESIS
1.7 RESEARCH OBJECTIVES
CHAPTER TWO FUNCTIONAL CHARACTERIZATION OF COTTON GAMYB62L, A NOVEL R2R3 TFIN TRANSGENIC ARABIDOPSIS
2.1 MATERIAL AND METHODS
2.1.1 Bioinformatic analysis of GaMYB62L gene
2.1.2 Plant transformation and screening of 35S: GaMYB62L Arabidopsis lines
2.1.3 Response of overexpressed GaMYB62L transgenic lines to drought and cold stress
2.1.4 Germination and root elongation assays
2.1.5 Measurements of transpirational Water loss rate and Relative water content
2.1.6 Analysis of proline content and chlorophyll contents
2.1.7 Stomatal opening rate determination in response to Abscisic acid treatment
2.1.8 Gene expression analysis of Markers genes by qRT-PCR
2.1.9 Statistical analysis of Data
2.2 RESULTS
2.2.1 GaMYB62L Sequence Characterization and Phylogenetic analysis
2.2.2 GaMYB62L overexpression positively modulates drought tolerance in transgenic linesof Arabidopsis
2.2.3 Analysis of proline and chlorophyll content in GaMYB62L transgenic lines
2.2.4 Constitutive expression of 35S:GaMYB62L in Arabidopsis up-regulates tolerance toABA, salt and osmotic stresses
2.2.5 Reduced leaf Stomatal Density, size and stomatal opening in 35S:GaMYB62L
2.2.6 Cold response of GaMYB62L-overexpressing Arabidopsis plants
2.2.7 Transcripts investigation of abiotic stress-responsive genes
2.3 DISCUSSION
CHAPTER THREE GAMYB85, NOVEL COTTON R2R3 TF TRANSFORMED IN ARABIDOPSIS HASIMPORTANT ROLE IN DROUGHT TOLERANCE
3.1 MATERIAL AND METHODS
3.1.1 Bioinformatic analysis of GaMYB85 gene
3.1.2 Transgenic plant construction and screening
3.1.3 Germination and root elongation assays
3.1.4 Drought and freezing tolerance assay
3.1.5 Measurements of transpirational WLR and RWC
3.1.6 Proline content and chlorophyll content
3.1.7 Stomata measurements and opening rates determination in response to Abscisic acidtreatment
3.1.8 Gene expression analysis of marker genes by qRT-PCR
3.1.9 Statistical analysis of Data
3.2 RESULTS
3.2.1 GaMYB85 characterization and phylogenetic analysis
3.2.2 Overexpression of 35S:GaMYB85 in plants confers ABA hypersensitivity for seedgermination and root elongation
3.2.3 Overexpression of 35S:GaMYB85 in plants confers enhanced salt and osmotictolerance
3.2.4 Enhanced GaMYB85 tolerance under drought and freezing stress
3.2.5 Reduced stomatal density, increased stomatal response, and increased stomata sizein GaMYB85 plants
3.2.6 Abiotic stress responsive marker genes transcript analysis
3.3 DISCUSSION
CHAPTER FOUR CONCLUSION
References
SUPPLEMENTARY DATA I
SUPPLEMENTARY DATA II
ACKNOWLEDGEMENT
RESUME
【参考文献】：
期刊论文
[1]Arabidopsis C3HC4-RING finger E3 ubiquitin ligase At AIRP4 positively regulates stress-responsive abscisic acid signaling[J]. Liang Yang,Qiaohong Liu,Zhibin Liu,Hao Yang,Jianmei Wang,Xufeng Li,Yi Yang. Journal of Integrative Plant Biology. 2016(01)
[2]Transgenic expression of MYB15 confers enhanced sensitivity to abscisic acid and improved drought tolerance in Arabidopsis thaliana[J]. Zhenhua Ding a, b, Shiming Li a, b, Xueli An a, Xin Liu a, Huanju Qin a, Daowen Wang a, a State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China b Graduate School of Chinese Academy of Sciences, Beijing 100039, China. 遗传学报. 2009(01)
[3]利用SSR标记技术研究棉属A、D染色体组的进化[J]. 郭旺珍,王凯,张天真. 遗传学报. 2003(02)
本文编号：3652859
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