植物特異蛋白激酶SnRK2s是逆境脅迫應(yīng)答信號(hào)傳遞的樞紐,不僅參與對(duì)各種逆境脅迫的應(yīng)答,還參與植物的生長發(fā)育和養(yǎng)分利用。本研究從小麥(Triticum aestivum L.)中克隆了基因TaSnRK2.9s。利用qRT-PCR技術(shù),分析小麥幼苗在逆境脅迫和ABA處理?xiàng)l件下TaSnRK2.9的表達(dá)模式;利用轉(zhuǎn)基因水稻驗(yàn)證TaSnRK2.9功能;根據(jù)TaSnRK2.9-5A序列的SNPs(single nucleotide polymorphisms)開發(fā)了CAPS(cleaved amplified polymorphic sequence)和KASP(kompetitive allele specific PCR)標(biāo)記,分析小麥群體的單倍型,以及其與農(nóng)藝性狀的關(guān)系。主要研究結(jié)果如下:i)基因表達(dá)模式分析發(fā)現(xiàn),TaSnRK2.9參與小麥對(duì)干旱、高鹽、冷脅迫及ABA處理的應(yīng)答;ii)過量表達(dá)TaSnRK2.9能促進(jìn)水稻幼苗根系生長,提高穗粒數(shù),增強(qiáng)細(xì)胞膜穩(wěn)定性,降低葉綠素a與葉綠素b的比值,提高抗旱性;iii)從小麥中克隆了TaSnRK2.9的三個(gè)拷貝,根據(jù)其染色體來源分別命名為TaSn...

【文章頁數(shù)】：106 頁

【學(xué)位級(jí)別】：博士

【文章目錄】：
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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