| A Method for Radioactive Labelling of Hebeloma cylindrosporum to Study Plant-fungus Interactions
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Author:
Date:
2017-10-20
[Abstract] In order to quantify P accumulation and P efflux in the ectomycorrhizal basidiomycete fungus Hebeloma cylindrosporum, we supplied 32P to mycelia previously grown in vitro in liquid medium. The culture had four main steps that are 1) growing the mycelium on complete medium with P, 2) transfer the mycelia into new culture solution with or without P, 3) adding a solution containing 32P and 4) rinsing the mycelia before incubation with or without plant. The main point is to rinse very carefully the mycelia after 32P supply in order to avoid overestimation of 32P efflux into the medium.
[摘要] 为了量化外生菌根担囊菌真菌Hebeloma cylindrosporum中的P积累和P流出，我们向以前在体外生长的菌液提供了 32 P 中。 培养物有四个主要步骤：1）在具有P的完全培养基上培养菌丝体，2）将菌丝体转移到具有或不具有P的新培养溶液中，3）加入含有32和32的溶液 ）在与或不与植物孵育之前冲洗菌丝体。 要点是在 32 P供应之后非常仔细地冲洗菌丝体，以避免过高估计P P。
【背景】众所周知，菌根真菌和植物之间的关联改善了宿主植物的P营养（由Smith和Read，2008; Plassard和Dell，2010; Cairney，2011; Smith等人，2015）。这种积极的作用主要是由于真菌细胞探索远离根部的土壤的磷酸盐（Pi）吸收，允许探索大量的土壤超过主要吸收根部周围形成的耗尽区（Smith and Read，2008; Cairney，2011; Smith ，2015）。然而，为了受益于宿主植物，吸收的Pi必须从探测土壤的真菌细胞转移到与宿主细胞紧密接触的细胞中。在外生菌根共生中，这些交流被认为发生在外生菌根内的“Hartig网”领域（Smith and Read，2008; ...
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| Determination of Rate of [3H-methyl]-choline Incorporation into Cellular Lipids and Non-lipid Metabolites
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Author:
Date:
2016-11-20
[Abstract] The choline-containing phospholipid, phosphatidylcholine (PtdCho) is the most common mammalian phospholipid found in cell membrane (Ide et al., 2013). It is also a component of intracellular signalling pathways (Cui and Houweling, 2002). Herein is described a measure of the rate of accumulation of choline by lipid soluble PtdCho and lyso-Ptdcho which can further be discriminated by chromatographic analysis (Smith and Phyu, 2016). Determination of the accumulation of [3H-methyl]-choline into water-soluble components is also described. The procedure could be used to measure the effect of drugs and other factors on choline incorporation into phospholipids. After exposure of cells to test conditions (e.g., drugs) adherent cells in tissue culture flasks are ...
[摘要] 含胆碱的磷脂，磷脂酰胆碱（PtdCho）是在细胞膜中发现的最常见的哺乳动物磷脂（Ide等人，2013）。它也是细胞内信号传导途径的组成部分（Cui和Houweling，2002）。本文描述了脂质可溶性PtdCho和溶血磷脂酰胆碱积累胆碱的速率的量度，其可通过色谱分析进一步区分（Smith和Phyu，2016）。还描述了[3 H] - 甲基] - 胆碱在水溶性组分中的积累的测定。该程序可用于测量药物和其它因素对胆碱掺入磷脂的影响。在将细胞暴露于测试条件（例如，药物）后，将组织培养瓶中的贴壁细胞与放射性标记的[3 H] - 甲基] - 胆碱在培养基中温育15分钟（脉冲）。然后快速除去[3 H] - 甲基] - 胆碱，并在非放射性培养基（Chase）存在下继续孵育。然后通过细胞分级和测量脂质和非脂质细胞组分中的放射性来测定[3 H] - 甲基的细胞分布。
[背景] 磷脂代谢在细胞膜形成中是必需的（Ide等人，2013）和细胞信号传导（Cui和Houweling， 2002）。包含胆碱的代谢物的形成和胆碱在脂质中的积累是细胞增殖过程中的关键过程。磷脂代谢紊乱与癌症和其他疾病有关（Gibellini和Smith，2010）。这些过程的测量对于理解医学成像模态至关重要，所述医学成像模式使用[11 C] - ...
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