Summary
Plants regenerated from tissue cultures of maize were screened for variants of ADH1 and ADH2. Root extracts of 645 primary regenerant plants were tested, and one stable mutant of Adh1 was detected. The mutant gene (Adh1-Usv) produces a functional enzyme with a slower electrophoretic mobility than that of the progenitor Adh1-S allele, and is stably transmitted to progeny. The mutant was not present among four other plants derived from the same immature embryo, and therefore arose as a consequence of the culture procedure. The gene of Adh1-Usv was cloned and sequenced. A single base change in exon 6 was the only alteration found in the gene sequence. This would translate in the polypeptide sequence to a valine residue substituting for a glutamic acid residue, resulting in the loss of a negative charge and the production of a protein with slower electrophoretic mobility.
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Abbreviations
- kb:
-
kilobase pairs
- ADH:
-
alcohol dehydrogenase
References
Ahloowalia BS (1982) Plant regeneration from callus culture in wheat. Crop Sci 22:405–410
Armstrong KC, Nakamura C, Keller WA (1983) Karyotype instability in tissue culture regenerants of triticale (x Triticosecale Wittmack) cv ‘Welsh’ from 6-month-old callus cultures. Z Pflanzenzüchtg 91:233–245
Beckert M, Pollacsek M, Caenen M (1983) Etude de la variabilité génétique obtenue chez le maïs après callogenèse et régénération de plantes in vitro. Agronomie 3:9–18
Brettell RIS, Pallotta MA, Gustafson JP, Appels R (1986) Variation at the Nor loci in triticale derived from tissue culture. Theor Appl Genet (in press)
Davies PA, Pallotta MA, Ryan SA, Scowcroft WR, Larkin PJ (1986) Somaclonal variation in wheat: genetic and cytogenetic characterisation of alcohol dehydrogenase 1 mutants. Theor Appl Genet, in press
Dennis ES, Gerlach WL, Pryor AJ, Bennetzen JL, Inglis A, Llewellyn D, Sachs MM, Ferl RJ, Peacock WJ (1984) Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize. Nucleic Acids Res 12:3983–4000
Dennis ES, Sachs MM, Gerlach WL, Finnegan EJ, Peacock WJ (1985) Molecular analysis of the alcohol dehydrogenase 2 (Adh2) gene of maize. Nucleic Acids Res 13:727–743
Edallo S, Zucchinali C, Perenzin M, Salamini F (1981) Chromosomal variation and frequency of spontaneous mutation associated with in vitro culture and plant regeneration in maize. Maydica 26:39–56
Ferl RJ, Dlouhy SR, Schwartz D (1979) Analysis of maize alcohol dehydrogenase by native-SDS two dimensional electrophoresis and autoradiography. Mol Gen Genet 169:7–12
Frischauf A-M, Lehrach H, Poustka A, Murray N (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842
Green CE, Phillips RL (1975) Plant regeneration from tissue cultures of maize. Crop Sci 15:417–421
Hanson AD, Brown AHD (1984) Three alcohol dehydrogenase genes in wild and cultivated barley: characterisation of the products of variant alleles. Biochem Genet 22:495–515
Karp A, Maddock SE (1984) Chromosome variation in wheat plants regenerated from cultured immature embryos. Theor Appl Genet 67:249–255
Landsmann J, Uhrig H (1985) Somaclonal variation in Solanum tuberosum detected at the molecular level. Theor Appl Genet (in press)
Larkin PJ, Scowcroft WR (1981) Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214
Larkin PJ, Ryan SA, Brettell RIS, Scowcroft WR (1984) Heritable somaclonal variation in wheat. Theor Appl Genet 67:443–455
McCoy TJ, Phillips RL (1982) Chromosome stability in maize (Zea mays) tissue cultures and sectoring in some regenerated plants. Can J Genet Cytol 24:559–565
McCoy TJ, Phillips RL, Rines HW (1982) Cytogenetic analysis of plants regenerated from oat (Avena sativa) tissue cultures; high frequency of partial chromosome loss. Can J Genet Cytol 24:37–50
Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either strand of double-digest restriction fragments. Gene 19:269–276
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Sachs MM, Dennis ES, Gerlach WL, Peacock WJ (1986) Two alleles of maize Alcohol dehydrogenase 1 have 3′ structural and poly(A) addition polymorphism. Genetics, in press
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schwartz D (1966) The genetic control of alcohol dehydrogenase in maize: gene duplication and repression. Proc Natl Acad Sci USA 56:1431–1436
Schwartz D, Endo J (1966) Alcohol dehydrogenase polymorphism in maize—simple and compound loci. Genetics 53:709–715
Sutton WD, Gerlach WL, Schwartz D, Peacock WJ (1984) Molecular analysis of Ds controlling element mutations of the Adh1 locus of maize. Science 223:1265–1268
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Wahl GM, Stern M, Stark GR (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxy-methylpaper and rapid hybridisation by using dextran sulphate. Proc Natl Acad Sci USA 76:3683–3687
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Communicated by P. Starlinger
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Brettell, R.I.S., Dennis, E.S., Scowcroft, W.R. et al. Molecular analysis of a somaclonal mutant of maize alcohol dehydrogenase. Mol Gen Genet 202, 235–239 (1986). https://doi.org/10.1007/BF00331642
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DOI: https://doi.org/10.1007/BF00331642