35S promoter-driven transgenes are variably expressed in different organs of Arabidopsis thaliana and in response to abiotic stress
35S promoter-driven transgenes are variably expressed in different organs of Arabidopsis thaliana and in response to abiotic stress
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DOI：
10.1007/s11033-021-06235-x
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发表时间：
2021-02-25
影响因子：
2.8
通讯作者：
Dubrovina, Alexandra S.
中科院分区：
文献类型：
--
作者：
Kiselev, Konstantin V.;Aleynova, Olga A.;Dubrovina, Alexandra S.
花椰菜花叶病毒(CaMV)35S启动子是植物生物技术中最常用的启动子。尽管CaMV 35S被广泛认为是一种具有较高转录活性的结构性启动子，但其转录稳定性还没有得到广泛的研究。本研究以模式植物拟南芥为材料，对两种广泛应用的转基因植物新霉素磷酸转移酶II(NPTII)和增强型绿色荧光蛋白(EGFP)进行了全面的表达分析，它们受CaMV 35S双启动子调控，依赖于器官类型、时间、植物年龄以及对非生物胁迫的响应。实时定量聚合酶链式反应(qRT-PCR)分析表明，在3个独立的转基因拟南芥品系中，NPTII和EGFP在子叶、幼叶和根中的转录水平显著高于花序、茎和成叶。NPTII和EGFP的表达在白天有变化，并随着植株年龄的增加而增加。干旱和低温胁迫对转基因的表达有显著影响，而高温、高盐度和伤害对转基因的表达没有显著影响。这项研究表明，由共同构成启动子驱动的转基因可以表现出显著的转录活性差异，这取决于植物的器官、生理条件和对非生物胁迫的响应。因此，在收获植物材料之前，应注意转基因植物材料和培养条件，以确保转基因活性的高和稳定。
The cauliflower mosaic virus (CaMV) 35S promoter is known as the most frequently used promoter in plant biotechnology. Although it is widely considered to be a strong constitutive promoter exhibiting high transcriptional activity, the transcriptional stability of CaMV 35S has not been extensively studied. Using the model plant species Arabidopsis thaliana, this study aimed for a comprehensive expression analysis of two widely used plant transgenes, neomycin phosphotransferase II (NPTII) and enhanced green fluorescent protein (EGFP), regulated by a double CaMV 35S promoter depending on the organ type, time of day, plant age, and in response to abiotic stress conditions. Quantitative real-time PCR (qRT-PCR) analysis revealed that the NPTII and EGFP transcript levels were markedly higher in the cotyledons, young leaves, and roots than in the inflorescences, stems, and adult leaves of three independent transgenic A. thaliana lines. The expression of NPTII and EGFP varied during the day and was elevated with the plant age. Drought and cold stress considerably affected the expression of the transgenes, while heat, high salinity, and wounding had no significant effect. This study shows that transgenes driven by a common constitutive promoter can exhibit marked variations in transcriptional activity depending on plant organ, physiological conditions, and in response to abiotic stress. Therefore, to ensure high and stable transgene activity, considerable attention should be given to the transgenic plant material and incubation conditions before harvesting the plant material.