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Procedure for whole mount fluorescence in situ hybridization of interphase nuclei on Arabidopsis thaliana
Author(s) -
Bauwens Serge,
Katsanis Katerina,
Montagu Marc,
Oostveldt Patrick,
Engler Gilbert
Publication year - 1994
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1994.6010123.x
Subject(s) - arabidopsis thaliana , biology , fluorescence in situ hybridization , in situ , in situ hybridization , fluorescence , labelling , botany , chemistry , chromosome , genetics , biochemistry , optics , messenger rna , organic chemistry , physics , gene , mutant
Summary A procedure for whole mount fluorescence in situ hybridization (FISH) on plant tissue is reported. The technique was demonstrated on seedlings and flowers of Arabidopsis thaliana L. with rDNA as a probe, labelled, both for direct and indirect detection. It was found that fixation in 1% formaldehyde yielded the best results with respect to morphology and hybridization efficiency. The combination of whole mount FISH and confocal scanning laser microscopy allowed the nuclear localization of the rDNA loci in all tissues of both seedlings and flowers. Direct labelling yielded the best signal‐to‐noise ratio, especially in the apical zones of the seedlings. The technique was further illustrated on seedlings of A. thaliana in double labelling experiments with rDNA and a tandemly repeated, 500 bp sequence of A. thaliana . Although nuclei in all tissues in the seedling exhibited both signals, hybridization efficiency for both signals was reduced in the dense, apical zones as compared with single labelling experiments with rDNA.