發(fā)布時間：2021-04-22 00:54

　　【目的】克隆甜椒內(nèi)質(zhì)網(wǎng)小分子熱激蛋白基因（CaHSP22.5）,并檢測其表達特性,為探究其在植物抗逆中的調(diào)控機制提供理論依據(jù)�！痉椒ā靠寺aHSP22.5基因并構(gòu)建其表達載體pBI121-CaHSP22.5,轉(zhuǎn)化農(nóng)桿菌LBA4404后通過葉盤法轉(zhuǎn)化野生型煙草,即獲得CaHSP22.5轉(zhuǎn)基因株系;以轉(zhuǎn)pBI121空載體的野生型煙草為對照組。對轉(zhuǎn)基因株系和野生型煙草進行4℃6 h低溫處理,根據(jù)幼苗成活率測定CaHSP22.5基因表達對植株耐寒性的影響。采用脈沖振幅調(diào)制葉室熒光儀Li-6400測量4℃處理10 h后野生型植株與CaHSP22.5轉(zhuǎn)基因株系葉綠素熒光,根據(jù)非光化學淬滅系數(shù)（NPQ）值測定CaHSP22.5基因表達對在低溫下植株光合作用的影響;使用過氧化氫（H2O2）鈦絡合物法對煙草葉片中H2O2進行定量分析,通過測定活性氧（ROS）積累程度分析CaHSP22.5對植物光合系統(tǒng)的影響;采用檢測檸檬酸合成酶（CS）活性的方法,在體外通過對變性CS復性的影響測定CaHSP22.5的分子伴侶活性。【結(jié)果】經(jīng)低溫處理后發(fā)現(xiàn)CaHSP22.5轉(zhuǎn)基因株系13號、24號和32號幼苗的平均存活... 

【文章來源】：南方農(nóng)業(yè)學報. 2020,51(05)北大核心CSCD

【文章頁數(shù)】：11 頁

【文章目錄】：
0 Introduction
1 Materials and methods
    1.1 Plant materials and growth conditions
    1.2 Isolation and sequencing of CaHSP22.5 gene from sweet pepper
    1.3 Vector construction and plant transformation
    1.4 Northern and western blotting analysis
    1.5 Seed germination
    1.6 Fluorescence measurements
    1.7 Analysis of H2O2and O
    1.8 Molecular chaperone activity assays
    1.9 Statistical analysis
2 Results and analysis
    2.1 Isolation and characterization of c DNA of Ca HSP22.5 gene from sweet pepper
    2.2 Expression of Ca HSP22.5 gene in sweet pepper during heat stress
    2.3 Enhanced chilling tolerance of transgenic tobacco over-expressing Ca HSP22.5 gene
    2.4 Enhancedphotosynthesisoftransgenictobacco overexpressing Ca HSP22.5 gene under chilling stress
    2.5 Reduced reactive oxygen species accumulation and alleviated lipid peroxidation in transgenic tobacco over-expressing Ca HSP22.5 gene under chilling stress
    2.6 Molecular chaperone activity of Ca HSP22.5in vitro
3 Discussion
4 Conclusion



本文編號：3152837