Staining and microscopy of mycorrhizal fungal colonization in preserved ericoid plant roots

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Kiheri , H , Heinonsalo , J & Timonen , S 2017 , ' Staining and microscopy of mycorrhizal fungal colonization in preserved ericoid plant roots ' , Journal of Berry Research , vol. 7 , no. 4 , pp. 231-237 . https://doi.org/10.3233/JBR-170160

Title: Staining and microscopy of mycorrhizal fungal colonization in preserved ericoid plant roots
Author: Kiheri, Heikki; Heinonsalo, Jussi; Timonen, Sari
Contributor organization: Department of Food and Nutrition
Department of Forest Sciences
Resarch Group of Annele Hatakka
Jussi Heinonsalo / Principal Investigator
Viikki Plant Science Centre (ViPS)
Ecosystem processes (INAR Forest Sciences)
Sari Timonen / Research Group
Forest Soil Science and Biogeochemistry
Teachers' Academy
Date: 2017-12
Language: eng
Number of pages: 7
Belongs to series: Journal of Berry Research
ISSN: 1878-5093
DOI: https://doi.org/10.3233/JBR-170160
URI: http://hdl.handle.net/10138/309890
Abstract: BACKGROUND: Visualization of ericoid mycorrhizal colonization using traditional methods relies on either fresh or KOH stored samples. Increasing interest in studying ericoid mycorrhization has highlighted a need for methods which can be used for preserved samples and are simple to implement with commonly available equipment. OBJECTIVE: The aim of this study was to improve on traditional techniques for staining ericoid mycorrhizal fungi and microscopically visualizing ericoid mycorrhizal roots which have been preserved. METHODS: Ericoid mycorrhizal roots were placed in KOH or frozen at -20 degrees C for long-term storage. Traditional Trypan Blue staining methods were modified to reduce damage to fine mycorrhizal hyphae and cortical cells. A high light-intensity dark-field microscopy technique was applied to clearly visualize stained mycorrhizae. The novel application was compared to other commonly used practices. RESULTS: Trypan Blue staining without KOH storage or clearing allowed for successful staining of ericoid mycorrhizal roots stored at -20 degrees C. The application of high light-intensity dark-field microscopy provided high contrast visualization of mycorrhizal structures. CONCLUSIONS: The modified Trypan Blue staining method was effective on frozen root samples, with dark-field microscopy being particularly effective at visualizing dark colored roots. Advantages to this method are lowcost and relatively fast application time. Therefore, this method is a realistic option for large scale analyses with many samples which require long-term preservation.
Subject: Ericoid
mycorrhizae
dark-field microscopy
Trypan Blue staining
mycorrhizal colonization
INOCULATION
1183 Plant biology, microbiology, virology
4112 Forestry
Peer reviewed: Yes
Rights: unspecified
Usage restriction: openAccess
Self-archived version: acceptedVersion


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