Quantification of the Composition Dynamics of a Maize Root-associated Simplified Bacterial Community and Evaluation of Its Biological Control Effect
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Author:
Date:
2018-06-20
[Abstract] Besides analyzing the composition and dynamics of microbial communities, plant microbiome research aims to understanding the mechanism of plant microbiota assembly and their biological functions. Here, we describe procedures to investigate the role of bacterial interspecies interactions in root microbiome assembly and the beneficial effects of the root microbiota on hosts by using a maize root-associated simplified seven-species (Stenotrophomonas maltophilia, Ochrobactrum pituitosum, Curtobacterium pusillum, Enterobacter cloacae, Chryseobacterium ...
[摘要] 除了分析微生物群落的组成和动态外,植物微生物群落研究的目的在于了解植物微生物群落组装的机制及其生物学功能。在这里,我们描述了通过使用与玉米根相关的简化的七种物种(嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia),)来研究细菌种间相互作用在根微生物群组装中的作用以及根菌群对宿主的有益影响的程序。 Ochrobactrum pituitosum ,Curtobacterium pusillum ,阴沟肠杆菌, Chryseobacterium indologenes ,<在我们以前的工作中描述过的假单胞菌(pseudomonas putida))合成细菌群落。表面灭菌的玉米种子在含有多种细菌的悬浮液中浸泡后,在基于双管生长室的非生物体系中生长。通过对7种菌株中的每种菌株选择性培养基的培养依赖性方法追踪定殖在玉米根部的细菌群组成的动态。通过监测社区结构的变化来评估细菌相互作用对社区组装的影响。通过量化(1)真菌植物病原体,种子表面上的fusarium="" verticillioides和(2)幼苗枯萎的严重程度来评估简化的7种群落的植物保护效果真菌引起的疾病,在细菌群落的存在和不存在的情况下。我们的方案将作为研究实验室条件下植物="" -=""> 在我们以前的工作中描述过的假单胞菌(pseudomonas> ...
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RNA Cap Methyltransferase Activity Assay
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Author:
Date:
2018-03-20
[Abstract] Methyltransferases that methylate the guanine-N7 position of the mRNA 5’ cap structure are ubiquitous among eukaryotes and commonly encoded by viruses. Here we provide a detailed protocol for the biochemical analysis of RNA cap methyltransferase activity of biological samples. This assay involves incubation of cap-methyltransferase-containing samples with a [32P]G-capped RNA substrate and S-adenosylmethionine (SAM) to produce RNAs with N7-methylated caps. The extent of cap methylation is then determined by P1 nuclease digestion, thin-layer chromatography (TLC), and phosphorimaging. ...
[摘要] 甲基化mRNA 5'帽结构的鸟嘌呤-N7位置的甲基转移酶在真核生物中普遍存在并且通常由病毒编码。这里我们提供生物样品的RNA帽甲基转移酶活性的生化分析的详细方案。该测定包括将含有帽 - 甲基转移酶的样品与[32 P] G-加帽的RNA底物和S-腺苷甲硫氨酸(SAM)温育以产生具有N7-甲基化帽的RNA。然后通过P1核酸酶消化,薄层色谱(TLC)和磷成像确定帽甲基化的程度。此处描述的方案包括用于产生[32 P] G-加帽的RNA底物和用于从哺乳动物细胞制备核和细胞质提取物的附加步骤。该分析也适用于分析其他生物样品(包括重组蛋白制剂和来自分析分离和免疫沉淀/下拉实验的级分)的帽甲基转移酶活性。
【背景】mRNA的5'端的N7-甲基鸟苷帽是适当的真核mRNA加工,定位和翻译所必需的修饰。 ...
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Fatty Acid Content and Composition of Triacylglycerols of Chlorella kessleri
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Author:
Date:
2018-01-05
[Abstract] Triacylglycerols (TAGs) are esters formed from one glycerol and three fatty acids. TAGs are induced to accumulate in algal cells under environmental stress conditions including nutrient-limitation, hyperosmosis, and low temperature, for the storage of metabolic energy and carbon, and also for the consumption of excess energy (e.g., Hirai et al., 2016; Hayashi et al., 2017). Beside their physiological significance, the commercial utilization of algal TAG has been expected for the production of biodiesel, the methyl esters of fatty acids, from the aspect of ...
[摘要] 三酰甘油(TAG)是由一种甘油和三种脂肪酸形成的酯。在包括营养限制,高渗和低温在内的环境胁迫条件下,TAG被诱导积累在藻细胞中,用于代谢能和碳的储存以及多余能量的消耗(例如, Hirai等人,2016; Hayashi等人,2017)。除了它们的生理意义之外,从碳中性概念的角度来看,藻类TAG的商业利用已经被期望用于生产生物柴油,即脂肪酸的甲酯。 TAG的量可以通过定量测定其构成脂肪酸来确定。该方案由以下三部分组成:首先是根据Bligh和Dyer(1959)的方法,使用有机溶剂,氯仿和甲醇从藻细胞中提取总脂质,其次是TAG的分离通过薄层色谱法(TLC)从其他脂质类别中分离出来,第三种是产生其组成型脂肪酸的甲基酯化衍生物,随后通过毛细管气相色谱法(GLC)对其进行定量。该协议改编自Sato和Tsuzuki(2011),用于绿藻的TAG分析, Chlorella kessleri 。
【背景】已经使用几种方法测定TAG的脂肪酸含量。简单而方便的方案包括用脂肪酶处理将TAG转化成甘油,随后通过酶产生与产生颜色或荧光的探针反应的产物测量甘油含量(McGowan等人,1983; ...
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