小麦蔗糖非发酵相关蛋白激酶2基因TaSnRK2.9的功能分析

发布时间：2023-08-09 18:57

　　植物特异蛋白激酶SnRK2s是逆境胁迫应答信号传递的枢纽,不仅参与对各种逆境胁迫的应答,还参与植物的生长发育和养分利用。本研究从小麦(Triticum aestivum L.)中克隆了基因TaSnRK2.9s。利用qRT-PCR技术,分析小麦幼苗在逆境胁迫和ABA处理条件下TaSnRK2.9的表达模式;利用转基因水稻验证TaSnRK2.9功能;根据TaSnRK2.9-5A序列的SNPs(single nucleotide polymorphisms)开发了CAPS(cleaved amplified polymorphic sequence)和KASP(kompetitive allele specific PCR)标记,分析小麦群体的单倍型,以及其与农艺性状的关系。主要研究结果如下:i)基因表达模式分析发现,TaSnRK2.9参与小麦对干旱、高盐、冷胁迫及ABA处理的应答;ii)过量表达TaSnRK2.9能促进水稻幼苗根系生长,提高穗粒数,增强细胞膜稳定性,降低叶绿素a与叶绿素b的比值,提高抗旱性;iii)从小麦中克隆了TaSnRK2.9的三个拷贝,根据其染色体来源分别命名为TaSn...

【文章页数】：106 页

【学位级别】：博士

【文章目录】：
摘要
abstract
CHAPTER I - Introduction
    1.1 Environmental stresses
        1.1.1 Water stress
        1.1.2 Heat stress
        1.1.3 Other abiotic stresses
    1.2 Abscisic acid
    1.3 Protein kinases in abiotic stress signaling
        1.3.1 Calcium dependent protein kinases (CDPK)
        1.3.2 Mitogen-activated protein kinases (MAPK)
        1.3.3 Sucrose non-fermenting 1 (SNF1)-related protein kinase
    1.4 Rice Os SAPK
    1.5 DNA markers and their application in wheat breeding
    1.6 Marker assisted evaluation of germplasm resources for plant breeding
        1.6.1 Linkage and association analysis
        1.6.2 Populations for association analysis
    1.7 Cross genome marker studies
    1.8 Functional markers
        1.8.1 Functional marker development to isolate candidate gene from wheat genome
        1.8.2 Grain trait related genes and their FMs
    1.9 The purpose of the present study
CHAPTER II - Gene expression profiling and transgenic rice characterization
    2.1 Materials and methods
        2.1.1 Cloning Ta Sn RK2.9 and sequence analysis
        2.1.2 Relative expression studies
        2.1.3 Application of different stresses and stage specific sample collection
        2.1.4 RNA extraction protocol
        2.1.5 c DNA synthesis
        2.1.6 Quantitative real time PCR protocol
        2.1.7 Plant materials and growth conditions
        2.1.8 Drought tolerance assay
        2.1.9 Cell membrane stability
    2.2 Results
        2.2.1 Molecular characterization of Ta Sn RK2.9
        2.2.2 Dynamic expression of Ta Sn RK2.9-5A and Ta Sn RK2.9-5B
        2.2.3 Morphological characteristics of Ta Sn RK2.9 in transgenic rice
        2.2.4 Physiological traits of transgenic and WT rice
    2.3 Discussion
        2.3.1 Ta Sn RK2.9 contains the typical characteristics of Sn RK2 subfamily
        2.3.2 Ta Sn RK2.9 assists in increasing root numbers and grain number
        2.3.3 Physiological changes in transgenic Ta Sn RK2.9 plants under drought conditions
CHAPTER III - Ta Sn RK2.9-5A is associated with grain related traits in common wheat
    3.1 Materials and methods
        3.1.1. Leaf DNA extraction protocol
        3.1.2. PCR product purification protocol
        3.1.3 DNA ligation
        3.1.4 Transformation
        3.1.5 Analyzing positive clones
        3.1.6 Analyzing positive clones by restriction digestion
        3.1.7 Plasmid extraction protocol
        3.1.8 Sanger sequencing
        3.1.9 Marker designing
        3.1.10 Plant materials and phenotyping
        3.1.11 Chromosomal location
        3.1.12 Isolation and sequencing of Ta Sn RK2.9
        3.1.13 Functional marker development
        3.1.14 Association analysis
    3.2 Results
        3.2.1 Genetic characterization of Ta Sn RK2.9
        3.2.2 Sequence polymorphism assays and marker development
        3.2.3 Association analysis of Ta Sn RK2.9-5A haplotypes and agronomic traits
        3.2.4 Geographic distribution of Ta Sn RK2.9-5A haplotypes in China, Pakistan and Europe .
        3.2.5 Selection of Ta Sn RK2.9-5A haplotypes in China, Pakistan and European wheat breeding process
    3.3 Discussion
        3.3.1 Marker assisted evaluation of germplasm resources for crop breeding
        3.3.2 Ta Sn RK2.9-5A sequence polymorphism and chromosomal location
        3.3.3 Functional marker development and genotyping in wheat; a polyploid crop
        3.3.4 Ta Sn RK2.9 possess typical features of Sn RK2 subfamily
        3.3.5 Importance and association of TKW and GPS with Ta Sn RK2.9-5A haplotypes
CONCLUSION
REFERENCES
附件
ACKNOWLEDGEMENT
CURRICULUM VITAE



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