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Microchambers and video-enhanced light microscopy for monitoring cellularevents in living hyphae of arbuscular mycorrhizal fungi

机译：微腔室和视频增强光学显微镜，用于监测丛枝菌根真菌活菌丝中的细胞事件

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摘要

Mycelial elongation and protoplasmic flow rate in vitro were monitored for germinated spores of Gigaspora rosea and Glomus caledonium respectively, growing on membranes in microchambers, by using a combination of time-lapse and video-enhanced light microscopy and image analysis. The microchambers allowed continuous observation of living mycelium over a period of several hours during which protoplasm flow and bidirectional movements of cellular organelles and particles were monitored in individual hyphae. Growth rate of G. rosea hyphae, calculated 8 days after germination, was 2.64 mum/min. Protoplasmic flow rate, measured on the basis of the movement of particles, ranged from 2.98 to 4.27 mum/s in living hyphae of G. caledonium. We showed that G. rosea, when growing in axenic culture in the absence of the host, ceased growth within 8 days of germination and underwent a process of protoplasm retraction from hyphal tips, leading to the formation of empty mycelial segments. A process of resource reallocation was inferred in spores of G. rosea showing multiple germination. Detailed developmental studies of living hyphae by using microchambers could provide useful information on spatio-temporal dimensions of cellular events occurring in arbuscular mycorrhizal fungi.
机译：通过使用延时和视频增强光学显微镜以及图像分析的组合，分别监测了玫瑰长臂猿（Gigaspora rosea）和Glomus caledonium的萌发孢子在体外的菌丝体伸长和原生质流速，这些孢子在微腔室的膜上生长。微腔室可以连续观察几个小时内的活菌丝体，在此期间可以监测单个菌丝的原生质流以及细胞器和颗粒的双向运动。发芽后8天计算的玫瑰蔷薇菌丝的生长速率为2.64mum / min。根据颗粒的运动测得的原生质流，在加里登霉的活菌丝中为2.98至4.27 mum / s。我们表明，玫瑰木在无宿主的情况下在无性培养中生长时，在发芽后8天内停止生长，并经历了从菌丝尖端撤回原生质的过程，导致形成了空的菌丝体节段。在显示出多重发芽的玫瑰花孢子孢子中推断出资源重新分配的过程。使用微腔室进行的详细的活菌丝发育研究可以提供有关丛枝菌根真菌中发生的细胞事件的时空维度的有用信息。

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