Hydrogen Peroxide Measurement in Arabidopsis Root Tissue Using Amplex Red
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Author:
Date:
2016-11-05
[Abstract] This protocol describes the measurement of hydrogen peroxide (H2O2) content in Arabidopsis root tissue by using the Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit. When root tissue is disrupted and resuspended in phosphate buffer, H2O2 is released from the cells. The obtained root extracts containing H2O2 can be mixed with a solution containing Amplex® Red reagent (10-acetyl-3,7-dihydrophenoxazine). In the presence of horseradish peroxidase, the Amplex® Red reagent reacts with H2O2 in a 1:1 stoichiometry. The resulting product is the red-fluorescent compound resorufin which can be detected fluorometrically or spectrophotometrically. Our protocol is based on ...
[摘要] 该方案描述了在拟南芥根组织中通过使用Amplex来测量过氧化氢(H 2 O 2 O 2)含量。 sup>红色过氧化氢/过氧化物酶测定试剂盒。当根组织被破坏并重新悬浮在磷酸盐缓冲液中时,H 2 O 2 O 2从细胞中释放。可以将获得的含有H 2 O 2 O 2的根提取物与含有Amplex Red试剂(10-乙酰基-3,7-二 - 。在辣根过氧化物酶的存在下,Amplex Red试剂与H 2 O 2以1:1的化学计量反应。所得产物是红色荧光化合物试卤灵,其可以通过荧光测定或分光光度法检测。我们的方案是基于Amplex ®红色过氧化氢/过氧化物酶测定试剂盒的手册,并描述了一步一步的程序,详细描述必要的控制和数据分析。我们还包括旨在帮助用户容易地用其自己的样品再现测定的方案,说明和实施例的修改。 [背景] 活性氧(ROS),例如H 2 O 2 O 2可以在该细胞是发育提示或应激条件的结果。在大量,ROS积累可能是有害的,通过造成细胞损伤。然而,增加的ROS产生也可以具有信号传导作用,并且作为控制下游细胞反应的第二信使。在植物中,对于许多非生物胁迫,例如干旱,盐度,温度胁迫和营养缺乏,已经显示ROS的信号传导作用(Mittler,2002; Mittler和Blumwald,2015; Xia等人)。 ,2015)。在我们最近的出版物Le ...
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Establishing a Biofilm Co-culture of Pseudomonas and Aspergillus for Metabolite Extraction
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Author:
Date:
2015-12-05
[Abstract] Filamentous fungi and bacteria form mixed-species biofilms in nature and diverse clinical contexts (Frey-Klett et al., 2011; Peleg et al., 2010). The interactions between fungi and bacteria, often mediated by secreted metabolites, have important ramifications for the biology of the interacting partners (Frey-Klett et al., 2011). This is particularly true for the bacterium Pseudomonas aeruginosa (P. aeruginosa) and the fungus Aspergillus fumigatus (A. fumigatus) which often reside in the same niche such as lungs of cystic fibrosis (CF) patients. Some studies have reported that co-infection with P. aeruginosa and A. fumigatus could lead to a decrease in lung function relative to their respective single species ...
[摘要] 丝状真菌和细菌在自然界和不同的临床背景中形成混合物种生物膜(Frey-Klett等人,2011; Peleg等人,2010)。真菌和细菌之间的相互作用,通常由分泌的代谢物介导,对相互作用的伴侣的生物学具有重要的影响(Frey-Klett等人,2011)。对于绿脓假单胞菌(铜绿假单胞菌)和真菌烟曲霉( A。fumigatus >),其常常位于相同的位置,例如囊性纤维化(CF)患者的肺。一些研究报道了与p的共感染。铜绿和 烟曲霉可能导致相对于它们各自的单一物种感染的肺功能降低(Amin等人,2010; Peleg等人,2010)。代谢物提取和分析允许表征多微生物生物膜中的特定微生物代谢物。该方案描述了如何在固体培养基上制备假单胞菌属 - 曲霉菌共培养生物膜以准备代谢物提取。
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