Quantification of Salicylic Acid (SA) and SA-glucosides in Arabidopsis thaliana
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Author:
Date:
2018-05-20
[Abstract] Homeostasis between the cytoplasmic plant hormone salicylic acid (SA) and its’ inactive, vacuolar storage forms, SA-2-O-β-D-glucoside (SAG) and SA-β-D-Glucose Ester (SGE), regulates the fine-tuning of defense responses to biotrophic pathogens in Arabidopsis thaliana. This protocol describes a simplified, optimized procedure to extract and quantify free SA and total hydrolyzable SA in plant tissues using a classical HPLC-based method.
[摘要] 细胞质植物激素水杨酸(SA)与其无活性的液泡形式,SA-2-葡萄糖苷(SAG)和SA-β-D-葡萄糖酯之间的稳态 (SGE)规定了拟南芥中对营养性病原体防御反应的微调。 该协议描述了一种简化的优化程序,使用传统的基于HPLC的方法提取和定量植物组织中的游离SA和总可水解SA。
【背景】SA(2-羟基苯甲酸)是植物激素,其在叶绿体中响应于病原体攻击而合成。然后输出到细胞质中,在细胞质中建立局部和系统获得性抗性(SAR)。在广义方案中,植物对生物营养性病原体的抗性被认为是通过SA信号传导介导的,而对于坏死性病原体的抗性受茉莉酸(JA)和乙烯(ET)控制。 SA和JA / ET信号通路相互作用。 SA积累到高浓度是有毒的并导致细胞和组织损伤。因此,大多数病原体诱导的SA被UDP-葡糖基转移酶(UGT)糖基化以形成亲水的,无毒的SAG和SGE(Noutoshi等人,2012; George Thompson等人, 2017)。然后将SAG和SGE隔离在液泡中,在那里它们形成水解成活性SA的可重复使用的来源。因此,植物组织中增加的总SA(SA + SAG / ...
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Analysis of Malondialdehyde, Chlorophyll Proline, Soluble Sugar, and Glutathione Content in Arabidopsis seedling
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Author:
Date:
2013-07-20
[Abstract] The protocol has four sub-protocols, which are about the measurement of malondialdehyde, chlorophyll proline, soluble sugar, and glutathione content, respectively, in Arabidopsis seedling by using spectrophotometer. These methods are simple, effective and reproducible, which will help the researchers who are not familiar with these approaches, quickly get reliable results.
[摘要] 该方案具有四个子协议,其分别是使用分光光度计在拟南芥苗中测量丙二醛,叶绿素脯氨酸,可溶性糖和谷胱甘肽含量。 这些方法简单,有效和可重复,这将有助于不熟悉这些方法的研究人员快速获得可靠的结果。
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Macromolecular Biosynthesis Assay for Evaluation of Influence of an Antimicrobial on the Synthesis of Macromolecules
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Author:
Date:
2013-06-20
[Abstract] One of the most compelling approaches in the discovery of novel antimicrobials is screening of natural sources. In our publication we report on the activity of a compound 8-hydroxyserrulat-14-en-19-oic acid (EN4), a diterpene isolated from the Australian plant Eremophila neglecta. We evaluate its applicability for treatment of implant-associated infections. A comprehensive analysis of the mechanism of action of EN4 against staphylococci revealed its membranolytic properties and a general inhibition of macromolecular biosynthesis, which was confirmed in a macromolecular biosynthesis assay and suggested a multitarget activity. The method used to investigate an influence of EN4 on the synthesis of peptidoglycan, RNA, DNA and proteins is based on precipitation of macromolecules with ...
[摘要] 在发现新的抗微生物剂中最引人注目的方法之一是筛选天然来源。在我们的出版物中,我们报告了一种化合物8-羟基胆固醇-14-烯-19-酸(EN4)(一种从澳大利亚植物中分离的二萜烯)的活性。我们评估其适用于植入物相关感染的治疗。对EN4对葡萄球菌的作用机制的综合分析揭示其膜分解性质和大分子生物合成的一般抑制,其在大分子生物合成测定中被证实并提出多靶点活性。用于研究EN4对肽聚糖,RNA,DNA和蛋白质的合成的影响的方法是基于大分子与三氯乙酸的沉淀。这些大分子由相应的[3 H] - 标记的前体合成。测量掺入和不掺入抗微生物剂的放射性,并反映测试化合物的作用模式。使用具有已知作用机制的抗生素作为对照。
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