Preliminary screening of photosynthetically related cold resistance genes in Mandragora chinghaiensis
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[目的]挖掘高海拔、低温度下生长的青海茄参(Mandragora chinghaiensis Kuang et A.M.Lu)中的光合相关抗寒基因.[方法]以0 ℃低温胁迫青海茄参为试验材料,通过光合生理试验寻找关键时间点,进而进行转录组学分析,筛选光合相关抗寒基因并进行实时荧光定量(qRT-PCR)分析.[结果]净光合速率、气孔导度、蒸腾速率、最大荧光、光系统Ⅱ最大光化学效率、光化学猝灭系数以及电子传递速率呈逐渐下降趋势,胞间CO2浓度、初始荧光和非光化学猝灭系数呈上升趋势,且根据光合参数及叶绿素荧光参数的变化综合分析出关键时间节点为12 h,根据转录组分析筛选出7个基因,通过qRT-PCR分析表明,CP43基因表达先上调后下调,整体呈上调趋势;CAB-8、CAB-11、CP24-10B、CH-02、FD-C2与PSI-D2基因表达先下调后上调,整体呈下调趋势.[结论]本研究结果为青海茄参筛选光合相关抗寒基因以及后续的转基因验证提供了理论支持,达到了筛选基因的主要目的.
[Objective]The present paper aimed to explore the photosynthesis-related cold-resistant genes in Mandragora chinghaiensis grown at high altitude and low temperature.[Method]Taking 0 ℃ low temperature stress Qinghai eggplant ginseng as the experimental material,the key time points were found through photosynthetic physiological experiments,and then transcriptomics analysis was carried out to screen photosynthetic related cold-resistant genes and perform real-time fluorescence quantitative analysis.[Result]The net photosynthetic rate,stomatal conductance,transpiration rate,maximum fluorescence,maximum photochemical efficiency of photosystem Ⅱ,photochemical quenching coefficient,and electron transfer rate showed a gradual downward trend,while the intercellular CO2 concentration,initial fluores-cence,and non-photochemical quenching coefficient showed an upward trend.Based on the comprehensive analysis of changes in photosyn-thetic and chlorophyll fluorescence parameters,the key time node was 12 hours.According to the analysis of transcriptome,seven genes were screened.Through qRT-PCR analysis,the expression of CP43 gene was up-regulated first and then down-regulated,showing an overall up-ward trend;The expression of CAB-8,CAB-11,CP24-10B,CH-02,FD-C2,and PS1-D2 genes was first down-regulated and then up-regu-lated,showing an overall downward trend.[Conclusion]The results of the study provided some theoretical support for the screening of photo-synthesis-related cold resistance genes and the subsequent transgenic verification,the main purpose of screening genes has been achieved.
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