| In vivo and in vitro 31P-NMR Study of the Phosphate Transport and Polyphosphate Metabolism in Hebeloma cylindrosporum in Response to Plant Roots Signals
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Author:
Date:
2018-08-20
[Abstract] We used in vivo and in vitro phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy to follow the change in transport, compartmentation and metabolism of phosphate in the ectomycorrhizal fungus Hebeloma cylindrosporum in response to root signals originating from host (Pinus pinaster) or non-host (Zea mays) plants. A device was developed for the in vivo studies allowing the circulation of a continuously oxygenated mineral solution in an NMR tube containing the mycelia. The in vitro studies were performed on fungal material after several consecutive treatment steps (freezing in liquid nitrogen; crushing with perchloric acid; elimination of perchloric acid; freeze-drying; dissolution in an appropriate liquid ...
[摘要] 我们使用体内和体外磷-31核磁共振（ 31 P-NMR）光谱来跟踪运输，分区和 外生菌根真菌 Hebeloma cylindrosporum 中的磷酸盐代谢响应来自宿主（ Pinus pinaster ）或非宿主（ Zea mays ）的根信号植物。 开发了一种用于体内研究的装置，其允许连续氧化的矿物质溶液在含有菌丝体的NMR管中循环。 在几个连续的处理步骤（在液氮中冷冻;用高氯酸压碎;消除高氯酸;冷冻干燥;在适当的液体培养基中溶解）后，对真菌材料进行体外研究。
【背景】 菌根真菌和植物之间的关联改善了宿主植物的P营养（Smith和Read，2008; Plassard和Dell，2010; Cairney，2011; Smith 等人，，2015）。这种积极效应主要归因于真菌菌丝对磷酸盐（Pi）的吸收，探测了在活跃吸收根周围的耗竭区以外的大量土壤（Smith和Read，2008; Cairney，2011; Smith et al。< em="">，2015）和真菌细胞分泌细胞外磷酸酶（Quiquampoix和Mousain，2005）。吸收的Pi部分地掺入磷酸化的代谢物，磷脂和核酸中，并且部分地浓缩成多磷酸盐（PolyP），其中它们构成液泡中的储存池（Ashford 等人，，1994）。该协议详述了一种装置，该装置允许通过 31 ...
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| 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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| Establishing a Symbiotic Interface between Cultured Ectomycorrhizal Fungi and Plants to Follow Fungal Phosphate Metabolism
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Author:
Date:
2017-10-20
[Abstract] In ectomycorrhizal plants, the fungal cells colonize the roots of their host plant to create new organs called ectomycorrhizae. In these new organs, the fungal cells colonize the walls of the cortical cells, bathing in the same apoplasm as the plant cells in a space named the ‘Hartig net’, where exchanges between the two partners take place. Finally, the efficiency of ectomycorrhizal fungi to improve the phosphorus nutrition of their host plants will depend on the regulation of phosphate transfer from the fungal cells to plant cells in the Hartig net through as yet unknown mechanisms. In order to investigate these mechanisms, we developed an in vitro experimental device mimicking the common apoplasm of the ectomycorrhizae (the Hartig net) to study the phosphorus metabolism in the ...
[摘要] 在外生菌根植物中，真菌细胞定植于其宿主植物的根部，以产生称为外生菌根的新器官。在这些新器官中，真菌细胞定居在皮质细胞的壁上，与称为“Hartig网”的空间中的植物细胞在同一质粒中沐浴，两地之间的交流发生。最后，外源菌根真菌提高其宿主植物的磷营养的效率将取决于通过尚未知的机制调节从真菌细胞到Hartig网中植物细胞的磷酸转移。为了研究这些机制，我们开发了一种体外实验装置，模拟外生菌根（Hartig网）的常见质粒来研究外生菌根真菌中的磷代谢。 当真菌细胞与寄主植物“Pinus pinaster”的植物细胞相关联时。该装置可用于监测从先前与 32 P-正磷酸盐一起培养的真菌的磷酸盐外排。
【背景】已知菌根真菌和植物之间的关联可以改善植物P营养（由Smith和Read，2008; Plassard和Dell，2010; Cairney，2011; ...
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