Abstract
Aminopterin-resistant cell lines of maize were isolated by two different procedures of callus selection and by plating suspension cultures on drugcontaining medium after mutagen treatment. Efficiencies of different methods of variant selection were compared. Four aminopterin-resistant cell lines were shown to be 10–40 times more resistant than the parental cell line, and they were also resistant to another folate analog, methotrexate. The results suggest that alterations in at least three different cell properties could be responsible for resistance; 1) increased dihydrofolate reductase activity, 2) altered aminopterin sensitivity of dihydrofolate reductase, and 3) reduced drug uptake. One of the resistant cell lines showed more than one alteration, but its resistance proved to be unstable. The results suggest that stable changes which may or may not be of genetic origin and also unstable physiological changes or a combination of both could lead to aminopterin resistance in maize cell cultures.
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Abbreviations
- AMPT:
-
aminopterin
- MTX:
-
methotrexate
- DHFR:
-
dihydrofolate reductase
- MNNG:
-
N-methyl-N′-nitro-N-nitrosoguanidine
- EMS:
-
ethylmethane sulfonate
References
Blakely, R.L. (1969) The biological reduction of pteridines, In: The biochemistry of folic acid and related compounds, pp. 139–187, Tatum, E.L., Neuberger, A., eds. John Wiley and Sons, New York
Brettell, R.I.S., Goddard, B.V.D., Ingram, D.S. (1979) Selection of Tms-cytoplasm maize tissue cultures resistant to Drechslera maydis T-toxin. Maydica 24, 203–213
Flintoff, W.F., Davidson, S.V., Simminovitch, L. (1976) Isolation and partial characterization of three methotrexate-resistant phenotypes from Chinese hamster ovary cells. Somat. Cell Genet. 2, 245–261
Gegenbach, B.G., Green, C.E., Donovan, C.M. (1977) Induction of selected pathotoxin resistance in maize plants regenerated from cell cultures. Proc. Natl. Acad. Sci. USA 74, 5113–5117
Gresshoff, G.M. (1979) Cycloheximide resistance in Daucus carota cell cultures. Theor. Appl. Genet. 54, 141–143
Hibberd, K.A., Walter, T., Green, C.E., Gengenbach, B.G. (1980) Selection and characterization of a feedback-insensitive tissue culture of maize. Planta 148, 183–187
Hillcoat, B.L., Blakely, R.L. (1966) Dihydrofolate reductase of Streptococcus faecalis. I. Purification and some properties of reductase from the wild strain and from strain A. J. Biol. Chem. 241, 2995–3001
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.L. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275
Maliga, P., Sz-Breznovitz, A., Marton, L. (1973) Streptomycinresistant plant from callus culture of haploid tobacco. Nature New Biol. 244, 29–30
Maliga, P., Lázár, G., Svab, Z., Nagy, F. (1976) Transient cycloheximide resistance in a tobacco cell line. Mol. Gen. Genet. 157, 291–296
Maliga, P. (1978) Resistant mutants and their use in genetic manipulation. In: Frontiers of plant tissue culture, pp. 381–392, Thorpe, R.A., ed. Univ. of Calgary, Canada, Int. Ass. for Plant Tissue Culture
Mastrangelo, I.A., Smith, H.H. (1977) Selection and differentiation of aminopterin resistant cells of Datura innoxia. Plant Sci Lett. 10, 171–179
Mathews, C.K., Huennekens, F.M. (1963) Further studies on dihydrofolate reductase. J. Biol. Chem. 238, 3436–3442
Müller, A.J., Grafe, R. (1978) Isolation and characterization of cell lines of Nicotiana tabacum lacking nitrate reductase. Mol. Gen. Genet. 161, 67–76
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Ohyama, K. (1976) A basis for bromodeoxyuridine resistance in plant cells. Environ. Exp. Bot. 16, 209–216
Reddy, V.A., Rao, N.J. (1976) Dihydrofolate reductase from soybean seedlings. Characterization of the enzyme purified by affinity chromatography. Arch. Biochem. Biophys. 174, 675–683
Schimke, R.T., Kaufman, R.J., Alt, F.W., Kellems, R.F. (1978) Gene amplification and drug resistance in cultured murine cells. Science 202, 1051–1055
Suzuki, N., Iwai, K. (1970) The occurrence and properties of dihydrofolate reductase in pea seedlings. Plant Cell Physiol. 11, 199–208
Widholm, J.M. (1974) Selection and characterization of biochemical mutants of cultured plant cells. In: Tissue culture and plant science, pp. 287–299, Street, H.E., ed. Academic Press, New York
Yurina, N.P., Qdintsova, M.S., Maliga, P. (1978) An altered chloroplast ribosomal protein in a streptomycin resistant tobacco mutant. Theor. Appl. Genet. 52, 125–128
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Research supported by the College of Agriculture and Life Sciences and by the Graduate School, University of Wisconsin Madison, Wis, USA
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Shimamoto, K., Nelson, O.E. Isolation and characterization of aminopterin-resistant cell lines in maize. Planta 153, 436–442 (1981). https://doi.org/10.1007/BF00394982
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DOI: https://doi.org/10.1007/BF00394982