A clearing technique for detecting the fungal endophyte Acremonium sp. in grasses. 1993

K W Hignight, and G A Muilenburg, and A J van Wijk
Research Department, Advanta Seeds West Inc., Albany, Oregon.

Leaf tissue of tall fescue Festuca arundinacea Schreb., hard fescue Festuca ovina L., red fescue Festuca rubra L. and perennial ryegrass Lolium perenne L. was stained with rose Bengal or aniline blue to detect the presence of the fungal endophyte Acremonium sp.. Specimens were cleared using methyl salicylate, an optical clearing agent, and viewed using bright field microscopy. Tissue was preserved as dried tissue or stored in 70% aqueous ethyl alcohol before staining and clearing. Tissue was observed at 2, 4 and 12 weeks following clearing to check for stain retention. Staining with rose Bengal was inferior to aniline blue when followed by the clearing agent methyl salicylate. Fungal mycelia stained lighter with rose Bengal and were more difficult to detect than mycelia stained with aniline blue. The results illustrate the usefulness of combining staining and methyl salicylate clearing for detecting fungal endophytes.

UI MeSH Term Description Entries
D006109 Poaceae A large family of narrow-leaved herbaceous grasses of the order Cyperales, subclass Commelinidae, class Liliopsida (monocotyledons). Food grains (EDIBLE GRAIN) come from members of this family. RHINITIS, ALLERGIC, SEASONAL can be induced by POLLEN of many of the grasses. Alopecurus,Arundo,Gramineae,Grasses,Imperata,Grass,Imperata cylindrica
D000164 Acremonium A mitosporic fungal genus with many reported ascomycetous teleomorphs. Cephalosporin antibiotics are derived from this genus. Cephalosporium,Acremoniums,Cephalosporiums
D013194 Staining and Labeling The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts. Histological Labeling,Staining,Histological Labelings,Labeling and Staining,Labeling, Histological,Labelings, Histological,Stainings

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