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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