棉花苹果酸脱氢酶基因家族的分子鉴定及其特征

发布时间：2021-11-26 04:38

　　苹果酸脱氢酶（MDH,EC 1.1.1.37）属于A类脱氢酶,它可以组成具有高度保守性的NAD（P）⁺依赖性的基因家族,可催化草酰乙酸盐和苹果酸盐的可逆转换。而苹果酸是植物细胞中重要的中间代谢物,在乙醛酸循环、三羧酸循环、葡萄糖合成、氨基酸合成以及氧化还原稳定性等细胞代谢途径中具有多种生物学功能。目前,关于棉花MDH基因家族少有研究。本研究对棉花MDH基因家族进行了全基因组鉴定及比较分析,阐明了该基因家族在棉花基因组内的组成以及在棉纤维不同生长时期的表达特征,为该基因家族在棉纤维细胞发育过程中的功能研究奠定了基础。在雷蒙德氏棉、亚洲棉和陆地棉基因组中共鉴定到51个MDH基因。多重序列比对结果显示棉花MDH基因可以分为五个亚组:线粒体MDH,过氧化物酶体MDH,质体MDH,叶绿体MDH和细胞质MDH。相同亚组内的MDH基因在基因结构、氨基酸序列以及功能结构域的保守基序方面具有相似的特征。染色体定位分析暗示基因片段复制事件在棉花MDH基因家族扩张过程中发挥了主要作用,而纯化选择在棉花MDH基因家族随后的进化过程中也起到了重要作用。进一步的表达分析结果显示陆地棉MDH基... 

【文章来源】：清华大学北京市 211工程院校 985工程院校 教育部直属院校

【文章页数】：108 页

【学位级别】：博士

【文章目录】：
摘要
Abstract
Chapter 1 Introduction
    1.1 Malate dehydrogenase gene families in plants
        1.1.1 Plant malate dehydrogenase gene families
        1.1.2 Plant malate dehydrogenase functions
        1.1.3 Research progresses of cotton malate dehydrogenase
    1.2 Cotton fibers
        1.2.1 Cotton fiber development
        1.2.2 Cotton fiber elongation mechanism
        1.2.3 Importance of malate dehydrogenase during fiber elongation
    1.3 Cotton genomes
        1.3.1 Molecular evolution of cotton genome
        1.3.2 Sequence date of cotton genome
    1.4 Research objectives and proposed significance
        1.4.1 Key problems
        1.4.2 Research purposes and contents
        1.4.3 Proposed significance
Chapter 2 Material and methods
    2.1 Biomaterial
    2.2 Bioinformatics analysis
    2.3 RACE PCR
    2.4 Real-time quantitative and semi-quantitative RT-PCR
    2.5 Subcellular localization analysis
    2.6 Analysis of protein expression
    2.7 MDH activity assay and determination of malate content
    2.8 Prokaryotic expression and purification of proteins
Chapter 3 Identification of cotton MDH gene family and their sequence features
    3.1 Introduction
    3.2 Identification of MDH gene family in G. raimondii and G. arboretum genomes
    3.3 Identification of MDH gene family in G. hirsutum genome
    3.4 Sequence characteristics of cotton MDH genes
    3.5 Conserved domain analysis of cotton MDH genes
    3.6 Summary
Chapter 4 Evolutionary relationship and expression profiles of MDH gene family
    4.1 Introduction
    4.2 Phylogenetic relationships of cotton MDH genes
    4.3 Chromosomal location and duplication analysis of cotton MDH genes
    4.4 Expression pattern of MDH genes in different tissues of allotetraploid cotton
    4.5 Putative regulatory cis-elements in MDH gene promoters
    4.6 Summary
Chapter 5 Molecular characterization of GhcMDH1 gene
    5.1 Introduction
    5.2 Molecular cloning of a GhcMDH1 gene
    5.3 GhcMDH1 sequence features and phylogenetic analysis
    5.4 Tissue expression and subcellular localization of GhcMDH1
    5.5 MDH enzymatic activity and malate content in cotton fiber cells
    5.6 Summary
Chapter 6 Discussion and conclusions
    6.1 Discussion
    6.2 Main conclusions
    6.3 Future prospects
References
Acknowledgement
Appendix I
Appendix II
Personal resume and published papers


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期刊论文
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