【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(dá)特性,為探究其在植物抗逆中的調(diào)控機(jī)制提供理論依據(jù)�！痉椒ā靠寺aHSP22.5基因并構(gòu)建其表達(dá)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(duì)照組。對(duì)轉(zhuǎn)基因株系和野生型煙草進(jìn)行4℃6 h低溫處理,根據(jù)幼苗成活率測定CaHSP22.5基因表達(dá)對(duì)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?zé)晒?根據(jù)非光化學(xué)淬滅系數(shù)（NPQ）值測定CaHSP22.5基因表達(dá)對(duì)在低溫下植株光合作用的影響;使用過氧化氫（H2O2）鈦絡(luò)合物法對(duì)煙草葉片中H2O2進(jìn)行定量分析,通過測定活性氧（ROS）積累程度分析CaHSP22.5對(duì)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(duì)變性CS復(fù)性的影響測定CaHSP22.5的分子伴侶活性�！窘Y(jié)果】經(jīng)低溫處理后發(fā)現(xiàn)CaHSP22.5轉(zhuǎn)基因株系13號(hào)、24號(hào)和32號(hào)幼苗的平均存活... 

【文章來源】：南方農(nóng)業(yè)學(xué)報(bào). 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(hào)：3152837