發(fā)布時(shí)間：2024-02-15 10:05

　　水稻(Oryza sativa L.)是世界上最重要的糧食作物之一,為世界一半以上的人口提供主要食物。然而,全球氣候的不斷變化給水稻生產(chǎn)帶來(lái)重大挑戰(zhàn),特別是干旱環(huán)境,這嚴(yán)重影響了世界經(jīng)濟(jì)和糧食安全。對(duì)水稻的抗逆性狀進(jìn)行遺傳改良是應(yīng)對(duì)這一狀況的主要策略之一。根據(jù)已有報(bào)道,水稻抗旱性狀是由多基因控制的,因此,多基因聚合的策略在理論上可以用于改善水稻的抗旱性。然而,目前這種策略仍然缺乏實(shí)驗(yàn)證據(jù)。在本研究中,我們通過(guò)MISSA(multiple-round in vivo sitespecific assembly)系統(tǒng)組裝了從水稻中分離的幾個(gè)干旱應(yīng)答基因,并且通過(guò)農(nóng)桿菌介導(dǎo)轉(zhuǎn)化法將聚合載體以及對(duì)應(yīng)的單基因超表達(dá)載體導(dǎo)入KY131水稻品種中。之后,我們鑒定了T1代轉(zhuǎn)基因植株中的目的基因轉(zhuǎn)錄水平和插入片段拷貝數(shù),并選取單拷貝、超量表達(dá)目的基因的轉(zhuǎn)基因家系和陰性轉(zhuǎn)基因材料(KY131-N)用于大田干旱脅迫的預(yù)實(shí)驗(yàn)和之后的進(jìn)一步實(shí)驗(yàn)中。在預(yù)實(shí)驗(yàn)期間,通過(guò)對(duì)葉片死亡程度和結(jié)實(shí)率的目測(cè),我們發(fā)現(xiàn)共表達(dá)兩個(gè)基因OsbZIP46CA1(編碼bZIP轉(zhuǎn)錄因子OsbZIP46的組成型激活形式)和SAPK6(編碼...

【文章頁(yè)數(shù)】：111 頁(yè)

【學(xué)位級(jí)別】：博士

【文章目錄】：
Abstract in English
Abstract in Chinese
List of Abbreviations
1 Introduction
    1.1 The origin of the research issue
    1.2 Literature review
        1.2.1 Responses of plant under abiotic stresses
            1.2.1.1 Responses and signal transduction of plants under abiotic stresses
            1.2.1.2 Regulatory network of gene expression under abiotic stresses
        1.2.2 The role of ABA on abiotic stress response in plant
        1.2.3 Genetic transformation in plants
        1.2.4 Progress of technology study for the introduction of multiple genes into transgenic plants
        1.2.5 Candidate genes for genetic transformation
            1.2.5.1 bZIP transcription factor-encoding genes
            1.2.5.2 SAPK protein kinase-encoding genes
    1.3 Research purpose and significance
2 Materials and methods
    2.1 Experimental materials
    2.2 Agrobacterium-mediated transformation
    2.3 DNA extraction and PCR positive test
    2.4 RNA extraction, reverse transcription and Real-time PCR
    2.5 Southern blot hybridization
    2.6 ABA sensitivity test of the transgenic plants
    2.7 Stress tolerance experiments of transgenic rice at the seedling stage
        2.7.1 Drought resistance experiment of transgenic rice at the seedling stage
        2.7.2 Temperature stress resistance experiments of transgenic rice at the seedling stage
    2.8 Drought resistance experiment of transgenic rice at the reproductive stage
    2.9 Determination of stress-related physiological indicators in rice
        2.9.1 Determination of water loss rate in rice leaves
        2.9.2 Determination of cell membrane permeability
        2.9.3 Measure content of malondialdehyde (MDA) in rice leaves
    2.10 RNA-Seq and analysis
3 Results
    3.1 Screening of candidate genes and stress resistance in the transgenic plants
        3.1.1 Identification the presence of candidate genes in the transgenic plants
        3.1.2 Expression level analysis of candidate genes in the transgenic plants
        3.1.3 Copy number analysis of the partial T1 transgenic plants
        3.1.4 Phenotypic pre-evaluation of transgenic materials in the field
    3.2 Increased ABA sensitivity of XL22 co-overexpression seedlings
    3.3 Enhanced drought resistance of the XL22 transgenic seedlings
    3.4 Enhanced drought resistance of the XL22 transgenic plants at the reproductive stage
    3.5 The XL22 transgenic lines improved heat and cold tolerance
    3.6 Enhanced oxidation resistance of XL22 plants under heat stress
    3.7 Transcriptome profiling of the XL22 transgenic plants
4 Discussion
    4.1 Co-overexpression of OsbZIP46CA1 and SAPK6 enhances drought tolerance
    4.2 Rice drought resistance evaluation indicators
    4.3 Co-overexpression of OsbZIP46CA1 and SAPK6 improves tolerance to temperature stress
    4.4 Application of multi-gene assembly strategy in improving stress resistance of rice
References
Appendices
    Appendix 1. Supplemental figures
    Appendix 2. Supplemental datas
    Appendix 3. Protocols
        Protocol 1. Plasmid DNA extraction
        Protocol 2. Extraction of total DNA using Plant DNAzol reagent
        Protocol 3. Southern blot hybridization
Resume
Acknowledgement



本文編號(hào)：3899578