Summary
DNA methylation has been associated with gene activity in differentiating and developing plant tissues. The objective of this study was to determine the involvement of methylation in the expression of a gene transferred into carrot (Daucus carota L.) tissues by particle bombardment. Expression of the Dc8-GUS gene construct in response to treatments with 5-azacytidine (S-azaC) and to in vitro methylation by methylases was investigated by histochemical assay of GUS activity. The 5-azaC treatment increased the frequency of Dc8-driven GUS expression in both calli and somatic embryos. The increase occurred with treatment either to E. coli containing the plasmid insert or to the carrot tissues before bombardment. GUS expression, increased by the 5-azaC treatment, was enhanced by ABA treatment of both calli and somatic embryos and was more prominent in the latter. Increased digestion of the 5-azaC-treated plasmid DNA with EcoRII suggested that demethylation had occurred. In vitro methylation of Dc8-GUS by methylases generally resulted in a lower frequency of GUS expression. SssI methylase completely inhibited GUS expression. The level of GUS expression was correlated with the extent of methylation of the plasmid.
Similar content being viewed by others
Abbreviations
- ABA:
-
Abscisic Acid
- 5-azaC:
-
5-azacytidine
- GUS:
-
β-glucuronidase
- Dc8:
-
carrot promoter
References
Brettell RIS, Dennis ES (1991) Mol Gen Genet 229:365–372
Creusot F, Acs G, Christman JK (1982) J Biol Chem 257:2041–2048
Goupil P, Hatzopoulos P, Franz G, Hempel FD, You R, Sung ZR (1992) Plant Mol Biol 18:1049–1063
Guiltinan MJ, Marcotte WR, Quatrano RS (1990) Science 250:267–271
Hatzopoulos P, Fong F, Sung ZR (1990) Plant Physiol 94:690–695
Jefferson RA, Kavanagh TA, Bevan MW (1987) EMBO J 6:3901–3907
Juttermann R, Li E, Jaenisch R (1991) Proc Natl Acad Sci USA 91:11797–11801
Kirchner G, Kinslow CJ, Bloom GC, Taylor DW (1993) Plant Mol Biol Rep 11:320–325
Klaas M, John MC, Crowell DN, Amasino RM (1989) Plant Mol Biol 12:413–423
Kunze R, Starlinger P, Schwarte D (1988) Mol Gen Genet 214:325–327
Murashige T, Skoog F (1962) Physiol Plant 15:473–497
Palmgren G, Mattson O, Okkels FT (1993) Plant Mol Biol 21:429–435
Santi DV, Garrett CE, Barr PJ (1983) Cell 33:9–10
Topfer R, Schell J, Steinbiss HH (1988) Nucleic Acids Res 16:8725
Van Slogteren GMS, Hooykaas PJJ, Schilperoort RA (1984) Plant Mol Biol 3:333–336
Vanyushin BF, Belozersky AN, Kokurina, NA, Kadirova DX (1968) Nature (London) 218:1066–1067
Wagner I, Capesius I (1981) Biochim Biophys Acta 654:52–56
Watson JC, Kaufman LS, Thompson WF (1987) J Mol Biol 193:15–26
Weber H, Graessmann A (1989) FEBS Lett 253:163–166
Author information
Authors and Affiliations
Additional information
Communicated by G. C. Phillips
Rights and permissions
About this article
Cite this article
Zhou, Y., Magill, J.M., Magill, C.W. et al. DNA methylation and Dc8-GUS transgene expression in carrot (Daucus carota L.). Plant Cell Reports 15, 815–818 (1996). https://doi.org/10.1007/BF00233146
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00233146