Abstract
DNA-binding fluorochromes are often used for vital staining of plant cell nuclei. However, it is not always sure whether the cells after staining still remain in living state. We chose several criteria to estimate the validity of real vital staining for sexual cell nuclei. These were: the cytoplasmic streaming in pollen tubes whose nuclei were stained, the simultaneous visualization of fluorochromatic reaction and nucleus staining in isolated generative cells, and the capability of isolated, prestained generative or sperm cells to fuse with other protoplasts. The results confirmed that 4′,6-diamidino-2-phenylindole (DAPI), Hoechst 33258 and mithramycin could be used as real vital stains, though their efficiency varied from case to case; among them DAPI showed best effect. The fluorescent vital staining technique offered a useful means foridentification and selection of heterokaryons in gametoplast manipulation studies.
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This study is supported by the National Natural Sciences Foundation of China.
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Appendices
Plate 1
Fluorescence vital staining of pollen tube nuclei in Z. graniflora. × 240. g. Generative cell. v. Vegetative nucleus. See Fig 1, Fig 2, Fig 3, Fig 4, Fig 5, Fig 6
Plate 2
Fluorescence vital staining of isolated generative cells in H. minor. × 1000. See Fig 7, Fig 8, Fig 9, Fig 10, Fig 11, Fig 12
Plate 3
Detection of DAPI- prestained gametoplasts during protoplast fusion. g. Generative cell or its nucleus, m. Microspore protoplast or its nucleus, p. Petal protoplast or its nucleus, s. Sperm cells. See Fig 13, Fig 14, Fig 15, Fig 16, Fig 17, Fig 18, Fig 19, Fig 20
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Yang, H., Wu, X., Mo, Y. et al. Fluorescent vital staining of plant sexual cell nuclei with DNA-specific fluorochromes and its application in gametoplast fusion. Cell Res 3, 121–130 (1993). https://doi.org/10.1038/cr.1993.13
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DOI: https://doi.org/10.1038/cr.1993.13
