MYB轉(zhuǎn)錄因子是非常著名的一類轉(zhuǎn)錄因子,它們主要參與脅迫響應(yīng)的基因互做和信號(hào)通路。MYB轉(zhuǎn)錄因子在氨基酸的N端含有一個(gè)保守的、具有53個(gè)氨基酸殘基的DNA結(jié)合結(jié)構(gòu)域—MYB結(jié)構(gòu)域,MYB結(jié)構(gòu)域可以折疊成螺旋-環(huán)-螺旋結(jié)構(gòu)。近期研究發(fā)現(xiàn)MYB結(jié)構(gòu)域也可位于MYB轉(zhuǎn)錄因子氨基酸的C端。依據(jù)DNA結(jié)合結(jié)構(gòu)域的不同,MYB蛋白可以進(jìn)一步分成四類:MYB-1R,R2R3-MYB,R1R2R3-MYB（MYB3R）和4-RMYB類蛋白。R1R2R3-MYB類蛋白廣泛的存在于單子葉植物和雙子葉植物中,棉花D基因組是目前為止含有R2R3-MYB基因最多的基因組（相較于擬南芥126個(gè)）,大約具有205個(gè)。R2R3-MYB基因參與許多脅迫響應(yīng),植物發(fā)育和細(xì)胞內(nèi)代謝過程。R2R3-MYB轉(zhuǎn)錄因子涉及植物對多種非生物脅迫的應(yīng)答,但是對棉花中R2R3-MYB基因參與干旱脅迫應(yīng)答的研究還比較少。而且,在干旱脅迫下的分子網(wǎng)絡(luò)和植物基因的功能仍然不是很清楚。在本研究中,我們克隆了兩個(gè)來自亞洲棉的R2R3-MYB基因GaMYB62L和GaMYB85,并將它們轉(zhuǎn)化于擬南芥中。這兩個(gè)基因的過表達(dá)擬南芥展現(xiàn)出耐受干旱的表型,... 

【文章頁數(shù)】：114 頁

【學(xué)位級(jí)別】：博士

【文章目錄】：
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


【參考文獻(xiàn)】：
期刊論文
[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.  遺傳學(xué)報(bào). 2009(01)
[3]利用SSR標(biāo)記技術(shù)研究棉屬A、D染色體組的進(jìn)化[J]. 郭旺珍,王凱,張?zhí)煺?  遺傳學(xué)報(bào). 2003(02)



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