Abstract
Two amino-acid auxotrophic cell clones ofHyoscyamus muticus, VA5 (His-) and VIIIB9 (Trp-), isolated in a previous experiment have been characterised quantitatively. Studies of growth in the presence and absence of histidine and tryptophan and an examination of dose-response relationships for the two amino acids confirmed the strict auxotrophy of both cell lines. No revertants to prototrophy were detected in either cell line after more than two years in culture. N-methyl-N′-nitro-nitrosoguanidine did not induce reversion in VA5 cell populations. Wild-type aggregates mixed in various combinations with VA5 and VIIIB9 cells could be recovered after plating in selective conditions. No cross-feeding was detected, either between wild-type and auxotrophic cells or between the auxotrophic lines themselves. Both variants were recloned by protoplast culture. All protoplast-derived clones were auxotrophic. The auxotrophic phenotypes behaved as recessive traits in protoplast-fusion experiments.
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Abbreviations
- MNNG:
-
N-methyl-N′-nitro-nitrosoguanidine
References
Beadle, G.W., Tatum, E.L. (1945)Neurospora-II. Methods of producing and detecting mutations concerned with nutritional requirements. Am. J. Bot.32, 678–686
Gavazzi, G., Nava-Racchi, M., Tonelli, C. (1975) A mutation causing proline requirement inZea mays. Theor. Appl. Genet.46, 339–345
Gebhardt, C., Schnebli, V., King, P.J. (1981) Isolation of biochemical mutants using haploid mesophyll protoplasts ofHyoscyamus muticus. II. Auxotrophic and temperature-sensitive clones. Planta153, 81–89
Horsch, R.B., King, J. (1982) Reconstitution/counter-selection for auxotroph isolation fromDatura cell suspension. In: Proceedings of the Vth International Congress for Plant Tissue Culture, pp. 449–450, Fujiwara, A., ed. Maruzen, Tokyo
Müller, A.J., Grafe, R. (1978) Isolation and characterisation of cell lines ofNicotiana 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
Nagata, T., Takebe, I. (1971) Plating of isolated tobacco mesophyll protoplasts on organic medium. Planta99, 12–20
Potrykus, I., Harms, C.T., Lörz, H. (1979) Callus formation from cell culture protoplasts of corn (Zea mays L.). Theor. Appl. Genet.54, 209–214
Shimamoto, K., King, P.J. (1982) Isolation of a histidine auxotroph ofHyoscyamus muticus during attempts to apply BUdR enrichment. Mol. Gen. Genet.189, 69–72
Sidorov, V.A., Menczel, L., Maliga, P. (1981) Isoleucine requiringNicotiana plant deficient in threonine deaminase. Nature (London)294, 87–88
Strauss, A., King, P.J. (1981) Analysis of plating technique for viability and drug-sensitivity determinations usingRosa cells. Physiol. Plant.51, 123–129
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For the previous paper in this series see Gebhardt et al. 1981
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Gebhardt, C., Shimamoto, K., Lázár, G. et al. Isolation of biochemical mutants using haploid mesophyll protoplasts ofHyoscyamus muticus . Planta 159, 18–24 (1983). https://doi.org/10.1007/BF00998809
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DOI: https://doi.org/10.1007/BF00998809