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itzhak itzhak
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Posts: 1
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8 years ago
Hey all,

I am doing research on fungal relationships in the north east united states. In order to properly continue my research, I am attempting to collect soil samples and stain ectomycorrhizae.
My questions are:
What is the most effective and easy stain to use? In what way can I use these stained slides to quantify the volume of ectomycorrhizae? To clarify, I will be collecting multiple samples from separate plots under different conditions and comparing.

Thanks
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wrote...
Staff Member
7 months ago
Staining Ectomycorrhizae involves specific techniques to visualize and study these fascinating fungal-root symbiotic structures. Here are some methods commonly used for staining and studying ectomycorrhizae:

Fluorescence Microscopy:

Early stages of ectomycorrhizal (EM) development in roots can be detected using fluorescence microscopy. This technique allows researchers to observe the intricate interactions between fungal hyphae and plant roots.

By using fluorescent dyes or probes, you can highlight specific structures within the EM, such as the Hartig net (a complex hyphal branching network)

Isolation Techniques:

EM fungi can be isolated from various sources, including roots, sclerotia, rhizospheres, and sexual spores (basidiospores).
To isolate EM fungi, researchers typically grow them on synthetic agar media. Nutrient agar plates, sterile distilled water, or freeze-drying of vegetative mycelium are common methods for maintaining EM cultures.
Aniline Blue Staining:

Aniline blue is a fluorescent dye that binds to chitin, a component of fungal cell walls.
By staining EM structures with aniline blue, you can visualize fungal hyphae and their interactions with plant roots under a microscope.

Confocal Microscopy:


Confocal microscopy provides high-resolution, three-dimensional images of EM structures.
It allows researchers to explore the spatial arrangement of fungal hyphae within the root tissues and assess their colonization patterns.

Remember that studying ectomycorrhizae is essential for understanding their ecological roles and interactions with plants. These techniques help uncover the hidden world of symbiosis beneath the soil!
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