水稻和玉米叶肉细胞以及维管束鞘细胞转录组分析

发布时间：2024-06-10 22:47

　　C₄植物的光合作用效率高于C₃途径的植物接近50%。为了将C₄途径引入C₃植物中,需要对调控光合作用的关键基因的分子作用机理进行研究,特别是了解这些基因叶肉细胞(Mc)和维管束鞘细胞(BSc)中差异表达。因此,在本实验中的研究目的是:⑴从水稻和玉米中获得均一Mc和BSc,以便通过利用LCM技术进行细胞特定RNA测序;⑵分析水稻作为C₃植物的特定Mc和BSc基因的差异表达;⑶分析玉米作为C₄植物的特定Mc和BSc基因的差异表达。水稻中已经鉴定了包括Mc和BSc总共35956个基因,其中BSc特有2068个,Mc特有5842个。参与光合作用的基因有:在Mc中特异性表达的OsSET13与OsSET26(Rubisco LS甲基转移酶底物结合域)、CAB2R/OsLhcb1.2与RCABP89/OsLhcb2(叶绿素A-B结合蛋白)、PsaH与PsbX(光系统组分)、未知功能CHP01589、K09122、OsPM8ATP基因,但在BSc中这些基因也往往存在微量表...

【文章页数】：86 页

【学位级别】：博士

【文章目录】：
摘要
abstract
Chapter1 INTRODUCTION
    1.1 Photosynthetic pathways
        1.1.1 C₃ pathway
        1.1.2 C₄ pathway
    1.2 Evolution of C₄ photosynthetic pathway
        1.2.1 Phylogenetic distribution
        1.2.2 Kranz anatomy
        1.2.3 Molecular regulation of C₄ pathways
    1.3 The C₄ Rice Project
        1.3.1 Rice
        1.3.2 The C₄ Rice Project – Overview
        1.3.3 Previous studies
    1.4 Laser capture microdissection technique
        1.4.1 LCM systems
        1.4.2 LCM applications on plants
    1.5 Aims
Chapter2 MATERIALS AND METHODS
    2.1 Plant materials
        2.1.1 Oryza sativa
        2.1.2 Zea mays
    2.2 LCM sample preparation
        2.2.1 Leaf sample collection and fixation
        2.2.2.Microwave processing
        2.2.3 Microtome sectioning
        2.2.4 LCM
    2.3 RNA Extraction and qualification
        2.3.1 RNA extraction
        2.3.2 RNA integrity testing
    2.4 RNA amplification and RNA-seq
        2.4.1 RNA amplification
        2.4.2 RNA-seq
Chapter3 Mc- and BSc-specific RNA of rice and maize
    3.1 Introduction
    3.2 Results and discussion
        3.2.1 Leaf collection
        3.2.2 Laser capture microdissection
        3.2.3 RNA Extraction and RNA Integrity Testing
    3.3 Discussion
Chapter4 Differential expression of rice Mc and BSc transcriptomes
    4.1 Introduction
    4.2 Results
        4.2.1 RNA-seq and alignment against the rice reference genome
        4.2.2 Differential expression of genes between rice BSc and Mc
    4.3 Discussion
        4.3.1 RNA-seq and alignment against the reference genome
        4.3.2. Differential expression of genes between BSc and Mc
Chapter5 Differential expression of maize Mc and BSc transcriptomes
    5.1 Introduction
    5.2 Results
        5.2.1 RNA-seq and alignment against the reference genome
        5.2.2 Differential expression of genes between BSc and Mc
    5.3 Discussion
        5.3.1 RNA-seq and alignment against the reference genome
        5.3.2 Differential expression of genes between BSc and Mc
CONCLUSION
References
致谢Acknowledgement
作者简历RESUME



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