I need guidance to know the method of isolation of mycorrhizal fungi from root tissues and soil from the workers who performed the same methods practically.
@ Gangwar, Phillips and Hayman (1970) developed the method to visualize AM fungi in roots using trypan blue in lactophenol, but the use of phenol is not discouraged (Koske and Gemma, 1989). The KOH is used as cleaning agent for non-pigmented roots and H2O2 or NaOCl may be used for pigmented roots. The colonized root length may be estimated by gridline-intersect method (Giovannetti and Mosse, 1980).
For estimation of number of spores in soil you may follow the method of Hyman (1984) or Schenck and Perez 1990), to my experience the wet sieving method is the best.
Dear G. P. Gangwar
You can isolate spores of mycorrhizal fungi from rhizosphere soil by wet sieving and decanting method. Good luck
Jawad
Please see:
Article Methods for culturing and isolating arbuscular mycorrhizal fungi
Article Isolation and characterisation of Arbuscular mycorrhizal (AM...
https://aem.asm.org/content/aem/57/2/434.full.pdf
Article Isolation of Native Arbuscular Mycorrhizal Fungi within Youn...
https://res.mdpi.com/d_attachment/agronomy/agronomy-10-00106/article_deploy/agronomy-10-00106.pdf
Book Working with Mycorrhizas in Forestry and Agriculture
Gonzalo Galileo Rivas Platero , Jawad K. Abood Al-Janabi Carine Temegne Nono J. C. Tarafdar Thanks to you all.....
We had success plating dispersed samples of soils in PDA agar plates supplemented with chloramphenicol (100 ug mL-1) and Rose Bengal (0.05 g L-1).
The question and its justification in replies was very helpful for my understanding as i am new to this field.
Through interpersonal channels (Facebook, Telluride Mushroom Festival), I can try to cross connect people as necessary:
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I initiated a conversation with Tradd Cotter of Mushroom Mountain via the Tellutide Mushroom Festival regarding fungi that might help process waste pinestraw (needles). This is to record some brief comments and links. BTW, what he knows of [fungi] has no edibility so their utility would be solely in waste remediation unless other info/ options exist.
Some notes I've recently collected via another source:
A technique/model for the propagation of native AM fungi https://rodaleinstitute.org/science/articles/how-to-innoculate-arbuscular-mycorrhizal-fungi-on-the-farm-part-1/
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This source was mentioned in the book Mycorrhizal Planet by Michael Philips, 2017, Chelsea Green publishing. https://www.chelseagreen.com/product/mycorrhizal-planet/
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I've been looking for a method (someone else's or my own) to propagate/ enhance and inoculate (PEI) forest ArM fungi taken from a healthy forest site (or "archived" from the subject site) into another with depleted or effectively "absent" fungal communities due to tree thinning, severe burns, climate change and other factors. Finding this excited me in that other researchers are already looking at and developing technologies for similar ecological needs.
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From the Rodale web article: >
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Also: >
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OK, this survey *article* has a number of references to what appear to be journal studies which might serve as sources for my journal article review as well as tracking a topic of interest in potential future work of my own.
>
Examining these journal articles is work TBD.
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Part 2 of the original article follows at https://rodaleinstitute.org/science/articles/how-to-innoculate-arbuscular-mycorrhizal-fungi-on-the-farm-part-2/
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Part 2 informally describes an experiment in PEI. It utilizes soil taken from 5 different types of native sites (8-10 samples each) on the farm site. This could augment the diversity of fungi being inoculated and could conceivably be modified for a forest mycorestoration site.
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At the end of Part 2: >
This is another source to follow up on.
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Dear G. P. Gangwar please take a look at INVAM website: https://invam.wvu.edu/
Wet sieving and decanting method is good method to isolate mycorrhizal fungi from roots and soil.
With regards!
Chapter 16 Wet-sieving and Decanting Techniques for the Extraction o...
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