The use of DNA-specific dyes, i. e. DAPI, is extensively reported for phytoplasma detection in fresh plant materials. However, fluorescence-based microscopy and imaging of fresh tissues often evidences technical limitations which are more significant in infected tissues, because phenolic and other defense-related compounds accumulate in the cell wall and in the vacuole making difficult sample preparation. In this paper we describe a method based on the use of epifluorescence microscopy and the DNA probes DAPI and SYTO13® for phytoplasma visualization in resin-embedded plant tissues. The method allows detection of phytoplasmas and it is recommended for tissues that are recalcitrant to conventional imaging.
Epifluorescence microscopy imaging of phytoplasmas in embedded leaf tissues using DAPI and SYTO13 fluorochromes
PAGLIARI, Laura;MUSETTI, Rita
2016
Abstract
The use of DNA-specific dyes, i. e. DAPI, is extensively reported for phytoplasma detection in fresh plant materials. However, fluorescence-based microscopy and imaging of fresh tissues often evidences technical limitations which are more significant in infected tissues, because phenolic and other defense-related compounds accumulate in the cell wall and in the vacuole making difficult sample preparation. In this paper we describe a method based on the use of epifluorescence microscopy and the DNA probes DAPI and SYTO13® for phytoplasma visualization in resin-embedded plant tissues. The method allows detection of phytoplasmas and it is recommended for tissues that are recalcitrant to conventional imaging.Pubblicazioni consigliate
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