| Extraction, Purification and Quantification of Diffusible Signal Factor Family Quorum-sensing Signal Molecules in Xanthomonas oryzae pv. oryzae
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Author:
Date:
2017-03-20
[Abstract] Bacteria use quorum-sensing (QS) systems to monitor and regulate their population density. Bacterial QS involves small molecules that act as signals for bacterial communication. Many Gram-negative bacterial pathogens use a class of widely conserved molecules, called diffusible signal factor (DSF) family QS signals. The measurement of DSF family signal molecules is essential for understanding DSF metabolic pathways, signaling networks, as well as regulatory roles. Here, we describe a method for the extraction of DSF family signal molecules from Xanthomonas oryzae pv. oryzae (Xoo) cell pellets and Xoo culture supernatant. We determined the levels of DSF family signals using ultra-performance liquid chromatographic system (UPLC) coupled with accurate mass ...
[摘要] 细菌使用群体感知（QS）系统监测和调节其人口密度。细菌QS涉及作为细菌通信信号的小分子。许多革兰氏阴性细菌病原体使用一类广泛保守的分子，称为扩散信号因子（DSF）家族QS信号。 DSF家族信号分子的测量对于了解DSF代谢途径，信号网络以及调节作用至关重要。在这里，我们描述了从Xanthomonas oryzae pv提取DSF家族信号分子的方法。 （ Xoo ）细胞沉淀和Xoo 培养上清液。我们使用超高效液相色谱系统（UPLC）与准确的质量飞行时间质谱仪（TOF-MS）联合测定DSF家族信号的水平。在UPLC / MS系统的帮助下，DSF的检测限低至1μM，大大提高了在细菌培养和反应混合物中检测DSF DSF家族信号分子的能力。
背景 黄单胞菌属 oryzae pv。米糠X ae））））））））））y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y y十二碳烯酸（DSF），顺式-2-十二碳烯酸（BDSF），顺式-10-甲基-2-十二碳烯酸（IDSF）和顺式<顺式 -11-甲基十二烷-2,5-二烯酸（CDSF），以调节毒力因子产生（图1）。 DSF家族信号的生物合成，感知和周转需要Xoo 中的rpf ...
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| Detection of Nitric Oxide and Determination of Nitrite Concentrations in Arabidopsis thaliana and Azospirilum brasilense
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Author:
Date:
2016-03-20
[Abstract] There is now general agreement that nitric oxide (NO) is an important and almost ubiquitous signal in plants. Nevertheless, there are still many controversial observations and differing opinions on the importance and functions of NO in plants. Partly, this may be due to the difficulties in detecting and quantifying NO. Here, we summarize protocols for detecting NO and quantifying nitrite concentration in Arabidopsis seedlings. We also present a method to measure NO in biofilms formed by the plant growth promoting rhizobacteria Azospirillum brasilense (A. brasilense). NO in oxygen-containing aqueous solutions has a short half-life that is often attributed to a rapid oxidation to nitrite. Here we detail the use of the fluorescent probe DAF-FM DA and the ...
[摘要] 现在普遍认为一氧化氮（NO）是植物中重要的和几乎普遍存在的信号。然而，对于植物中NO的重要性和功能仍有许多有争议的观察和不同的观点。部分地，这可能是由于检测中的困难以及甚至更多的在定量NO。在这里，我们总结了在拟南芥幼苗中检测NO和定量亚硝酸盐浓度的方案以及在由植物生长促进根瘤菌（Azospirillum brasilense）形成的生物膜中的NO实时测量（ > A。brasilense ）。含氧水溶液中的NO具有短的半衰期，其通常归因于快速氧化成亚硝酸盐。在这里我们详细的使用荧光探针DAF-FM DA和电化学方法分别直接检测和定量NO，Griess试剂通过其氧化的亚硝酸盐形式间接检测NO。这些方案可用于多种细胞类型和植物的不同组织以及微生物。
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