Summary
Mesophyll protoplasts isolated fromPetunia hybrida were subjected to UV radiation (280–360 nm) in an attempt to assess whether (a) UV radiation has an effect on cortical microtubule organization, (b) UV radiation affects the progression of protoplasts through the cell cycle, and (c) there is a connection between the effect of UV radiation on cell division and the polymerization state of the microtubules. The proto plasts were irradiated with the following UV doses: 4, 8, 12, and 24mmol photons/m2, 30 min after isolation. Cell cycle analysis and immuno-localization of microtubules were carried out 0, 24, 48, and 72 h after irradiation. The length of cortical microtubules was determined after irradiation and in corresponding controls. We found that UV radiation induced breaks in cortical microtubules resulting in shorter fragments with increasing dose. Also, the protoplasts were delayed in their progression through the cell cycle, with G1 and G2 phases being affected as well as the S phase. The commencement of DNA synthesis in the irradiated protoplasts followed the re-establishment of a microtubule network. At 48 h after irradiation the protoplasts in all treatments, except for the 24 mmol/m2, had cortical microtubules of similar length, and at 72 h after irradiation only the protoplasts that had received 24 mmol photons/m2 had not started dividing.
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Abbreviations
- BSA:
-
bovine serum albumin
- DMSO:
-
dimethyl sulfoxide
- FDA:
-
fluorescein diacetate
- MT:
-
microtubules
- MTSB:
-
microtubule stabilizing buffer
- PAR:
-
photosynthetically active radiation (400–700 nm)
- PBS:
-
phosphate buffered saline
- UV:
-
ultraviolet
References
Bergounioux C, Perennes C, Brown SC, Gadal P (1988 a) Cytometric analysis of growth-regulator-dependent transcription and cellcycle progression inPetunia protoplast cultures. Planta 175: 500–505
— — —, Sarda C, Gadal P (1988 b) Relationship between protoplast division, cell cycle stage and nuclear chromatin structure. Protoplasma 142: 127–136
Bootsma D, Humphrey RM (1968) The progression of mammalian cells through the division cycle following ultraviolet irradiation. Mutation Res 5: 289–298
Bornman JF (1989) Target sites of UV-B radiation in photosynthesis of higher plants. J Photochem Photobiol [B] 4: 145–158
Carlson JG (1976 a) Mitotic effects of monochromatic ultraviolet radiation at 225, 265 and 280 nm on eleven stages of the cell cycle of the grasshopper neuroblast in culture. 1. Overall retardation from the stage irradiated to nuclear membrane breakdown. Radiat Res 68: 57–74
— (1976 b) Mitotic effects of monochromatic ultraviolet radiation at 225, 265, and 280 nm on eleven stages of the cell cycle of the grasshopper neuroblast in culture. 2. Changes in the progression rate and cell sequence between the stage irradiated and nuclear membrane breakdown. Radiat Res 68: 75–83
Conia J, Bergounioux C, Perennes C, Muller P, Brown SC, Gadal P (1987) Flow cytometric analysis and sorting of plant chromosomes fromPetunia hybrida protoplasts. Cytometry 8: 500–508
Djordjevic B, Tolmach LJ (1967) Responses of synchronous populations of HeLa cells to ultraviolet irradiation at selected stages of the generation cycle. Radiat Res 32: 327–346
Domon M, Rauth AM (1968) Ultraviolet irradiation of mouse L cells: effects on DNA synthesis and progression through the cell cycle. Radiat Res 35: 350–368
Hahne G, Hoffman F (1984) Dimethyl sulphoxide can initiate cell divisions of arrested callus protoplasts by promoting cortical microtubule assembly. Proc Natl Acad Sci USA 81: 5449–5453
Hughes K, Forer A, Wilson P, Leggiardo C (1988) Ultraviolet microbeam irradiation of microtubules in vitro. The action spectrum for local depolymerization of marginal band microtubules in vitro matches that for reducing birefringence of chromosomal spindle fibres in vivo. J Cell Sci 91: 469–478
Olsen WM, Kirkhus B (1989) The epidermal cell kinetic response to ultraviolet B irradiation combines regenerative proliferation and carcinogen associated cell cycle delay. Photochem Photobiol 50: 391–397
Sillers PJ, Forer A (1983) Action spectrum for changes in spindle fibre birefringence after ultraviolet microbeam irradiations of single chromosomal spindle fibres in crane-fly spermatocytes. J Cell Sci 62: 1–25
Sokal RR, Rohlf FJ (1982) Biometry. W. H. Freeman, New York
Wick SM, Seagull RW, Osborn M, Weber K, Gunning BS (1981) Immunofluorescence of organized microtubule arrays in structurally stabilized meristematic plant cells. J Cell Biol 89: 685–690
Wilson PJ, Forer A (1988) Ultraviolet microbeam irradiation of chromosomal spindle fibres shears microtubules and permits study of the new free ends in vivo. J Cell Sci 91: 455–486
Zamansky GB, Chou I-N (1987) Environmental wavelengths of ultraviolet light induce cytoskeletal damage. J Invest Dermatol 89: 603–606
Zaremba TG, LeBon TR, Millar DB, Smejkal RM, Hawley RJ (1984) Effects of ultraviolet light on the in vitro assembly of microtubules. Biochemistry 23: 1073–1080
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Staxén, I., Bergounioux, C. & Bornman, J.F. Effect of ultraviolet radiation on cell division and microtubule organization inPetunia hybrida protoplasts. Protoplasma 173, 70–76 (1993). https://doi.org/10.1007/BF01378863
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DOI: https://doi.org/10.1007/BF01378863