Notes1. Caution: DAPI is labeled as a nontoxic product, but the toxicological properties of this reagent have not been fully investigated according to its Material Saferty Data Sheet (http://www.sigmaaldrich.com). We recommend using gloves while handling your samples.2. We recommend taking samples wich are less affected by disease, preferably from areas that do not show fully developed symp-toms, with the aim of avoiding false positives due to secondary infections by other microorganisms in the affested tissues.3. this step reduces the dehydration of the samples.4. We recommend having one or two layers of cells for a better fit of the samples on the slide after the DAPI stain.5. We recommend keeping the samples in dark to extend the stain's life in the prepared sample, because light reduces the quality of the samples and tends to remove the stain from them. For further observations, it is recommended to seal the samples, e.g., using a layer of vaseline. if you do not have a special slide sealer, nail polish works very well as a sealer.6. If more information about the fluorescence microscopy proce-dure is required, we reco,,ended reading variouns reviews, e.g.(24). Samples that contain phytoplasmas show hight DAPI dluorescence in the sieve tubes due to the specifis combination of this stain with DnA, whereas no fluorescence is seen in healthy plants (Fig,2). Generally, small points and aggregations near the cell wall of the phloem tissue cells are observed. This same procedure can be used to quantify phytoplasma-like bodies in phoem cell of infected plants (Fig.3).
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