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Bi‐fluorescence imaging for estimating accurately the nuclear condition of Rhizoctonia spp.
Author(s) -
Guermache F.,
RodierGoud M.,
Caesar A.,
Héraud C.,
Bon M.C.
Publication year - 2012
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2012.03245.x
Subject(s) - dapi , rhizoctonia , fluorescence , autofluorescence , fluorescence lifetime imaging microscopy , rhizoctonia solani , biology , botany , optics , physics , genetics , staining
Aims: To simplify the determination of the nuclear condition of the pathogenic Rhizoctonia , which currently needs to be performed either using two fluorescent dyes, thus more costly and time‐consuming, or using only one fluorescent dye, thus less accurate. Methods and Results: A red primary fluorescence (autofluorescence) of the hyphal cell walls and septa of Rhizoctonia spp. with green excitation is evidenced in Rhizoctonia spp. This property is exploited and combined for the first time with a conventional DAPI fluorescence to accurately determine the nuclear condition of Rhizoctonia . This bi‐fluorescence imaging strategy depicted the nuclear condition in Rhizoctonia spp. more accurately than the conventional DAPI fluorescence used alone and was validated against isolates previously genotyped by DNA sequencing. Conclusions: We demonstrated that the bi‐fluorescence imaging strategy was safe, accurate and simple to perform and interpret. Significance and Impact of the Study: The developed bi‐fluorescence imaging strategy provides a sensitive tool for determining the nuclear condition of Rhizoctonia strains. Its simplicity is a key advantage when there are numerous cultures to be examined.