AI Article Synopsis
- Fluorescent dyes were tested for their effectiveness in staining the viable hyphae of Neotyphodium lolii and N. coenophialum endophytes associated with certain grasses.
- Among the dyes used, CMFDA showed the best staining intensity and contrast with plant cells, making it particularly effective in visualizing the fungi.
- CMFDA also demonstrated resistance to photo bleaching, maintaining fluorescence for up to 2 hours after being excited, while none of the dyes managed to stain hyphae within seeds.
Article Abstract
Fluorescent dyes were assessed for their ability to stain viable hyphae of the fungi Neotyphodium lolii and N. coenophialum, symbiotic endophytes of the Pooideae grasses Lolium perenne and Festuca arundinacea, respectively. The fluorescein-based fluorophores; fluorescein diacetate (FDA), 5(6)-carboxy-fluorescein diacetate (CFDA), 5-chloromethylfluorescein diacetate (CMFDA) and the chitin-binding stain, Calcofluor while M2R, were assessed for staining of endophyte hyphae in vitro from axenic fungal cultures and in planta, including epidermal leaf sheath peels, nodes, ovaries, embryos and meristems. CMFDA produced the greatest intensity of staining of fungal hyphae and gave excellent contrast in planta compared to the plant cells. Compared to the other dyes, CMFDA was also the least affected by photo bleaching and continued to fluoresce up to 2 h after initial excitation. None of the fluorescent dyes stained fungal hyphae in seed.
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| http://dx.doi.org/10.3852/12-062 | DOI Listing |
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