NDLI: Microwave-assisted technology for the clearing and staining of arbuscular mycorrhizal fungi in roots

Content Provider	Springer Nature Link
Author	Dalpé, Yolande Séguin, Sylvie Marie
Copyright Year	2013
Abstract	The use of microwave irradiation as a source of energy to clear and stain intra-radical arbuscular mycorrhizal fungi propagules has been tested on a variety of indigenous and cultivated herbaceous plants. The aim of the study was to evaluate the efficiency of microwave irradiation on root softening, fungi tissue staining, and preservation of DNA integrity for subsequent molecular analyses. The proposed methodology has been adapted from the standard procedures used to detect and quantify mycorrhizal root colonization levels. Using a domestic microwave oven, tissue clearing and staining required together between 30 s and 1.5 min of microwave treatment to be completed, depending the diameter size of the roots. The well-performing chemical stains tested were acid fuchsin, trypan blue, and aniline blue. The acid fuchsin clearing and staining processes, as performed, were also demonstrated to preserve DNA integrity for further molecular analyses. Irradiation by microwaves has been used with success in our laboratory within the frame of several studies. It offers considerable time saving over traditional method, reducing processing times from several hours to a few minutes while decreasing considerably the amount of chemicals and energy required to perform analyses.
Starting Page	333
Ending Page	340
Page Count	8
File Format	PDF
ISSN	09406360
Journal	Mycorrhiza
Volume Number	23
Issue Number	4
e-ISSN	14321890
Language	English
Publisher	Springer-Verlag
Publisher Date	2013-01-23
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Irradiation Root colonization Root clearing Root staining Microbiology Plant Sciences Ecology Agriculture Forestry
Content Type	Text
Resource Type	Article
Subject	Genetics Plant Science Medicine Molecular Biology Ecology, Evolution, Behavior and Systematics

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