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DOTAP,一种脂质体转染试剂,引发拟南芥植物防御反应。

DOTAP, a lipidic transfection reagent, triggers Arabidopsis plant defense responses.

机构信息

Instituto de Biología Molecular Y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas (IBR-CONICET) and Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Ocampo y Esmeralda, 2000, Rosario, Argentina.

Laboratory of Structure and Function of Biological Membranes, Université Libre de Bruxelles, Boulevard du Triomphe, 1050, Brussels, Belgium.

出版信息

Planta. 2019 Feb;249(2):469-480. doi: 10.1007/s00425-018-3014-7. Epub 2018 Sep 25.

Abstract

DOTAP triggers Arabidopsis thaliana immunity and by priming the defense response is able to reduce bacterial pathogen attack. DOTAP is a cationic lipid widely used as a liposomal transfection reagent and it has recently been identified as a strong activator of the innate immune system in animal cells. Plants are sessile organisms and unlike mammals, that have innate and acquired immunity, plants possess only innate immunity. A key feature of plant immunity is the ability to sense potentially dangerous signals, as it is the case for microbe-associated, pathogen-associated or damage-associated molecular patterns and by doing so, trigger an active defense response to cope with the perturbing stimulus. Here, we evaluated the effect of DOTAP in plant basal innate immunity. An initial plant defense response was induced by the cationic lipid DOTAP in the model plant Arabidopsis thaliana, assessed by callose deposition, reactive oxygen species production, and plant cell death. In addition, a proteomic analysis revealed that these responses are mirrored by changes in the plant proteome, such as up-regulation of proteins related to defense responses, including proteins involved in photorespiration, cysteine and oxylipin synthesis, and oxidative stress response; and down-regulation of enzymes related to photosynthesis. Furthermore, DOTAP was able to prime the defense response for later pathogenic challenges as in the case of the virulent bacterial pathogen Pseudomonas syringae pv. tomato. Disease outcome was diminished in DOTAP-pre-treated leaves and bacterial growth was reduced 100 times compared to mock leaves. Therefore, DOTAP may be considered a good candidate as an elicitor for the study of plant immunity.

摘要

DOTAP 触发拟南芥的免疫反应,并通过启动防御反应,能够减少细菌病原体的攻击。DOTAP 是一种阳离子脂质,广泛用作脂质体转染试剂,最近它被确定为动物细胞固有免疫系统的一种强激活剂。植物是固着生物,与具有先天和获得性免疫的哺乳动物不同,植物只具有先天免疫。植物免疫的一个关键特征是能够感知潜在的危险信号,就像微生物相关、病原体相关或损伤相关的分子模式一样,通过这种方式,触发主动防御反应来应对干扰刺激。在这里,我们评估了 DOTAP 在植物基础先天免疫中的作用。通过阳离子脂质 DOTAP 在模式植物拟南芥中诱导初始植物防御反应,通过胼胝质沉积、活性氧产生和植物细胞死亡来评估。此外,蛋白质组学分析表明,这些反应与植物蛋白质组的变化相吻合,例如与防御反应相关的蛋白质上调,包括参与光呼吸、半胱氨酸和氧化脂素合成以及氧化应激反应的蛋白质;以及与光合作用相关的酶下调。此外,DOTAP 能够为后来的致病性挑战(如毒力细菌病原体丁香假单胞菌 pv.番茄)引发防御反应。与对照叶片相比,在 DOTAP 预处理叶片中,疾病结局减少,细菌生长减少了 100 倍。因此,DOTAP 可以被认为是研究植物免疫的一个很好的诱导剂候选物。

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