棉花AAI基因家族分析和陆地棉GhAAI66参与BR信号通路调控早花和生物量的研究

发布时间：2021-06-19 12:53

　　植物生长发育是从营养生长到诱导花器官发育生殖生长的过程。在植物中,由含有AAI（α淀粉酶抑制剂）结构域基因形成的基因家族尚未见报道。本研究中,我们从9个不同物种中共鉴定到336个AAI基因,其中陆地棉（Gossypium hirsutum L.）中包含122个。在整个植物界,AAI基因家族成员的氨基酸序列在进化过程中均高度保守。并且,伴随着显著的多倍体化和基因组重复现象,系统发育分析将AAI基因分为五个主要分支。本研究中,我们分别鉴定到由于部分或者全基因组重复产生的42个旁系同源基因对和216个直系同源基因对,说明基因重复在棉花AAI基因家族扩增中发挥重要作用。其中,GhAAI66在花器官中特异表达,并且响应植物激素处理。并且,拟南芥中异源过表达GhAAI66,以及VIGS沉默棉花GhAAI66结果显示,GhAAI66能诱发植物生长发育时期的转变,进而出现早花现象。另外,RNA-seq数据的GO和KEGG分析以及qRT-PCR验证结果表明,GhAAI66通过参与包括GA、JA和开花调控过程在内的多重开花信号途径诱导拟南芥早花的级联反应。因此,AAI家族基因的鉴定分析和功能验证为棉花育种... 

【文章来源】：中国农业科学院北京市

【文章页数】：83 页

【学位级别】：博士

【文章目录】：
摘要
ABSTRACT
ABBREVIATIONS
CHAPTER ONE INTRODUCTION
    1.1 Cotton
    1.2 AAI gene family:
    1.3 Brassinosteroids(BRs)
    1.4 Early Flowering
    1.5 Biomass
    1.6 Hypothesis
    1.7 Research Objectives
CHAPTER TWO Genome-wide identification of GhAAI genes and GhAAI66 triggers a phase transition to induce early flowering
    2.1 MATERIALS AND METHODS
        2.1.1 Gene identification and analysis of conserved residues
        2.1.2 Phylogenetic analysis and determination of protein motif distribution and gene structure
        2.1.3 Chromosomal location,gene duplication,and synteny analysis
        2.1.4 Vector construction and generation of transgenic lines
        2.1.5 RNA-seq,KEGG,and GO analysis
        2.1.6 Plant material,hormone treatments,and qRT-PCR analysis
    2.2 RESULTS
        2.2.1 Identification and analysis of conserved amino acid residues in AAIs
        2.2.2 Phylogenetic analysis of AAI genes
        2.2.3 Protein motifs and gene structure analysis
        2.2.4 Chromosomal location,gene duplication,and synteny analysis of GhAAI genes
        2.2.5 Tissue specific expression pattern and responses to phytohormone treatment
        2.2.6 Ectopic expression of GhAAI66 regulates phase transition to induce early flowering
        2.2.7 GhAAI66 integrates multiple flower signaling pathways to induce early flowering
    2.3 DISCUSSION
        2.3.1 AAI genes were highly conserved during evolution
        2.3.2 Expansion and duplication of GhAAI gene family during evolution
        2.3.3 GhAAI genes have ubiquitous expression in tissues and regulated hormone treatments
        2.3.4 GhAAI66 regulates early flowering in Arabidopsis
CHAPTER THREE NGhAAI66 mediates BR signaling and increase biomass
    3.1 MATERIALS AND METHODS
        3.1.1 Plant material and Fox hunting for bri1-5 suppressor
        3.1.2 Construction of expression vectors
        3.1.3 Plant treatment and hypocotyl measurement
        3.1.4 Generation of promoter-GUS transgenic plants
        3.1.5 Biomass analysis
        3.1.6 Gene expression by qRT-PCR and statistical analysis
    3.2 RESULTS
        3.2.1 Identification of GhAAI66 gene
        3.2.2 GhAAI66 expressed ubiquitously
        3.2.3 GhAAI66 is a dominant genetic suppressor of bri1-
        3.2.4 GhAAI66 mediate multiple aspects of plant growth and development in Arabidopsis
        3.2.5 GhAAI66 plays positive role in BR signaling pathway
        3.2.6 GhAAI66 is the direct target of GhBES
        3.2.7 GhAAI66 enhance vegetative biomass and seed size in Arabidopsis
    3.3 DISCUSSION
        3.3.1 GhAAI66 is a dominant genetic suppressor of bri1-5 and act downstream of BRI1 in BR signaling
        3.3.2 A functional BRI1 is required for GhAAI66 to mediates plant growth and development
        3.3.3 GhAAI66 functions through GhBES1 to regulate BR signaling pathway
        3.3.4 GhAAI66 enhance vegetative capacity and seed size in Arabidopsis
CHAPTER FOUR CONCLUSION
REFERENCES
SUPPLEMENTARY DATA
ACKNOWLEDGEMENT
CURRICULUM VITAE


【参考文献】：
期刊论文
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