bHLH转录因子ABP7-Like在玉米叶片和籽粒发育中的功能鉴定和分子机制分析

发布时间：2021-07-11 23:07

　　玉米（Zea mays L.）是世界上仅次于小麦和水稻的第三大谷类作物。玉米具有多种用途,如用作人类食品,动物饲料,制药以及工业产品。由于世界人口日益增加,但由于耕地面积有限,粮食短缺正成为目前的主要问题。因此,为了满足未来对玉米的需求,我们必须提高粮食作物的产量。叶片会通过光合作用影响籽粒产量,而玉米的最终产量取决于籽粒性状,因此鉴定与叶片和籽粒发育相关的因素对于提高玉米产量非常重要。本实验室利用酵母单杂的方法筛选玉米cDNA文库分离了能与ABRE（G-Box）顺式元件结合的bHLH转录因子ABP7。在拟南芥中过表达ABP7导致角果变大、种子大小和种子重量的增加。通过生物信息学分析,我们发现了ABP7有一个具有89%DNA和97%氨基酸序列相似性的同源基因,并将新鉴定的基因命名为ABP7-Like。本研究的重点是ABP7-Like在叶片和籽粒发育过程中的功能和分子机制解析。通过与其他已知bHLH转录因子的序列比较,发现ABP7-Like是含有保守bHLH区域的bHLH转录因子。研究表明ABP7-Like是一种转录激活因子,其能与ABRE顺式元件结合,并能够与ABP7形成异二聚体。AB... 

【文章来源】：中国农业科学院北京市

【文章页数】：131 页

【学位级别】：博士

【文章目录】：
摘要
abstract
LIST OF ABBRIVATION
CHAPTER 1 INTRODUCTION
    1.1 EARLY LEAF DEVELOPMENTAL EVENTS
        1.1.1 Hormone Signaling and Transcription Factor Networks
        1.1.2 Definition of the Dorso-Ventral Axis of the Leaf
        1.1.3 Control of leaf growth by Cell expansion
    1.2 FACTORS EFFECTING LEAF SIZE
    1.3 LEAF SENESCENCE
    1.4 PROGRAMMED CELL DEATH DURING LEAF SENESCENCE
    1.5 TRANSCRIPTIONAL REGULATION OF PLANT LEAF SENESCENCE
    1.6 HORMONE SIGNALING DURING LEAF SENESCENCE
    1.7 ROS AND LEAF SENESCENCE
    1.8 ROLE OF BHLH TRANSCRIPTION FACTORS IN LEAF SENESCENCE
    1.9 STRUCTURE OF MAIZE KERNEL
        1.9.1 Embryo
        1.9.2 Coleorhiza
    1.10 MAIZE KERNEL DEVELOPMENTAL STAGES
    1.11 CORN GRAIN FILL STAGES
    1.12 FACTORS AFFECTING ON SEED SIZE
        1.12.1 Transcription factors in early maize kernel development stages
        1.12.2 The bHLH Transcription factor Family and their role in maize kernel development
    1.13 SCOPE AND OUTLINE OF THIS THESIS
CHAPTER 2 MATERIALS AND METHODS
    2.1 MATERIALS
        2.1.1 Plant materials
        2.1.2 Bacterial and yeast strains
        2.1.3 Constructs used for this thesis(Stored in our Lab)
        2.1.4 Instruments used for this study
        2.1.5 Biochemicals
    2.2 MEDIA AND SOLUTIONS
    2.3 BUFFERS AND REAGENTS
    2.4 PROTOPLAST TRANSFORMATION ASSAY PROTOCOL
    2.5 YEAST ONE-HYBRID PROTOCOL
    2.6 CLONING OF GENE AND VECTOR CONSTRUCTION
    2.7 KERNEL PHENOTYPING
    2.8 GRAIN FILLING RATE MEASUREMENT
    2.9 GUS STAINING METHOD
    2.10 GUS FLUORESCENT QUANTIFICATION
    2.11 PARAFFIN TECHNIQUE FOR HISTOLOGICAL STUDY OF THE GUS STAINED TISSUES
    2.12 SAMPLE PREPARATION FOR RNA-SEQ
    2.13 TRANSCRIPTOME SEQUENCING AND ANALYSIS
    2.14 SDS-PAGE
    2.15 PROTOCOL
    2.16 WESTERN BLOT
    2.17 CHIP-SEQ
    2.18 LEAF CHLOROPHYLL ANALYSIS
    2.19 3,3-DIAMINOBENZIDINE(DAB),NITROBLUE TETRAZOLIUM CHLORIDE(NBT)STAINING
CHAPTER 3 RESULTS
    3.1 IDENTIFICATION AND CHARACTERIZATION OF ABP7-LIKE
        3.1.1 Identification of ABP7-Like
        3.1.2 Transcriptional activation activity analysis of ABP7-Like
        3.1.3 DNA binding activity analysis of ABP7-Like
    3.2 EXPRESSION PATTERN ANALYSIS OF ABP7-LIKE DURING THE VEGETATIVE AND REPRODUCTIVE STAGES
    3.3 FUNCTIONAL AND MOLECULAR MECHANISM ANALYSIS OF ABP7-LIKE TRANSCRIPTION FACTOR IN LEAF DEVELOPMENT AT VEGETATIVE STAGE
        3.3.1 Phenotypes of the transgenic plants during vegetative growth
        3.3.2 Changes in ROS accumulation and chlorophyll content in the transgenic plants during vegetative growth
        3.3.3 Transcriptome analysis of ABP7-Like-OE,ABP7-Like-RNAi,ABP7-Like-SRDX and ABP7-Like-VP64 leaves
    3.4 FUNCTIONAL AND MOLECULAR MECHANISM ANALYSIS OF ABP7-LIKE TRANSCRIPTION FACTOR AT REPRODUCTIVE STAGE
        3.4.1 The effect of ABP7-Like on kernel traits
        3.4.2 The effect of ABP7-Like on grain filling rate
        3.4.3 Transcriptome analysis of ABP7-Like-OE,ABP7-Like-RNAi,ABP7-Like-SRDX kernels
        3.4.4 ChIP-Seq Analysis of genome-wide DNA-binding sites of ABP7-Like
        3.4.5 Validation of potential direct target genes of ABP7-Like
CHAPTER 4 DISCUSSION
CHAPTER 5 CONCLUSION AND PROSPECT
    5.1 CONCLUSION
    5.2 PROSPECT
REFERENCES
APPENDICES
ACKNOWLEDGEMENT
RESUME


【参考文献】：
期刊论文
[1]Regulation of Leaf Senescence and Crop Genetic Improvement[J]. Xiao-Yuan Wu,Ben-Ke Kuai,Ji-Zeng Jia,Hai-Chun Jing.  Journal of Integrative Plant Biology. 2012(12)



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