Summary
Alcohol dehydrogenase (ADH) activity is expressed in Arabidopsis seeds and tissue cultures. During the germination process, ADH activity declines rapidly and is no longer detectable in 9- to 10-day-old seedlings. The synthesis of ADH could be demonstrated in seedlings submitted to anaerobiosis by 35S-methionine incorporation studies.
Callus, induced from seeds or leaves on a 2,4-dichlorophenoxyacetic acid (2,4-D)-containing medium, and cell suspension cultures are characterized by a high level of ADH activity. The incorporation of 35S-methionine and two-dimensional electrophoresis indicated that ADH induction was due to de novo synthesis of the polypeptides. In vitro translation of total poly (A)+-RNA from seedlings and callus showed that only callus mRNA was able to direct the synthesis of ADH polypeptides. This demonstrates the de novo synthesis of ADH mRNA during callus induction.
Northern blot hybridization, using in vitro labelled ADH1-F DNA from maize as a probe, revealed sequence homology at the mRNA level between Arabidopsis and maize.
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Aerts M, Jacobs M, Hernalsteens J-P, Van Montagu M, Schell J (1979) Induction and in vitro culture of Arabidopsis thaliana crown gall tumours. Plant Sci Lett 17:43–50
Aviv H, Leder P (1972) Purification of biologically active globin mRNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69:1408–1412
Balazs E, Lebeurier G (1981) Arabidopsis is a host of Cauliflower mosaic virus. Arab Inf Serv 18:130–134
Bennet MD, Smith JB (1976) Nuclear DNA amounts in angiosperms Proc R Soc Lond [Biol] 274:227–274
Bevan M, Northcote DH (1981) Subculture-induced protein synthesis in tissue cultures of Glycine max and Phaseolus vulgaris. Planta 152:24–35
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bidning. Anal Biochem 72:248–254
Brooker JD, Tomaszewski M, Marcus A (1978) Preformed messenger RNAs and early wheat embryo germination. Plant Physiol 61:145–149
Ciriacy M (1975) Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. I. Isolation and genetic analysis of adh mutants. Mutat Res 29:315–326
Chamberlain JP (1979) Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem 98:132–135
Danno G (1977) Isoelectric focusing of proteins separated by SDS-polyacrylamide gel electrophoresis. Anal Biochem 83:189–193
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
Dolferus R, Jacobs M (1984) Polymorphism of alcohol dehydrogenase in Arabidopsis thaliana (L) Heynh.: genetic and biochemical characterization. Biochem Genet 22:817–838
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
Ferl RJ, Brennan MD, Schwartz D (1980) In vitro translation of maize ADH: evidence for the anaerobic induction of mRNA. Biochem Genet 18:681–691
Freeling M (1973) Simultaneous induction by anaerobiosis or 2,4-D of multiple enzymes specified by two unlinked genes: differential Adh1 and Adh2 expression in maize. Mol Gen Genet 127:215–227
Freeling M, Schwartz D (1973) Genetic relationships between the multiple alcohol dehydrogenases of maize. Biochem Genet 8:27–36
Freeling M, Cheng DSK (1978) Radiation-induced alcohol dehydrogenase mutants in maize following allyl alcohol selection of pollen. Genet Res 31:107–129
Freeling M, Birchler JA (1981) Mutants and variants of the alcohol dehydrogenase-1 gene in maize. In: Setlow JW, Hollaender A (eds). Genetic engineering, principles and methods, vol 3. Plenum Press, New York, p 223
Gerlach WL, Pryor AJ, Dennis ES, Ferl RJ, Sachs MM, Peacock WJ (1982) cDNA cloning and induction of the alcohol dehydrogenase gene (Adh1) of maize. Proc Natl Acad Sci USA 79:2981–2985
Giles AB, Grierson D, Smith H (1977) In vitro translation of mRNA from developing bean leaves. Evidence for the existence of stored mRNA and its light-induced mobilisation into polysomes. Planta 136:31–36
Goldberg DA (1980) Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene. Proc Natl Acad Sci USA 77:5794–5798
Hake S, Kelley P, Taylor WC, Freeling M (1984) Molecular characterization of maize anaerobic genes and identification of aldolase. In preparation
Jacobs M, Dolferus R (1983) Analysis of the ADH system of Arabidopsis and its potentialities for transformation. Abstracts of the Fifteenth Miami Winter Symposium: Advances in gene technology, p 83
Jenkins NA, Kaumeyer JF, Young EM, Raff RA (1978) A test for masked message: the template activity of messenger ribonucleoprotein particles isolated from sea urchin eggs. Dev Biol 63:279–298
Lehrach H, Diamond D, Wozney JM, Boedtker H (1977) RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16:4743–4751
Leutwiler LS, Hough-Evans BR, Meyerowitz EM (1984) The DNA of Arabidopsis thaliana. Mol Gen Genet 194:15–23
Lutstorf U, Megnet R (1968) Multiple forms of alcohol dehydrogenase in Saccharomyces cerevisiae. I. Physiological control of ADH-2 and properties of ADH-2 and ADH-4. Arch Biochem Biophys 126:933–944
Martin C, Northcote DH (1981) Qualitative and quantitative changes in mRNA of castor beans during the initial stages of germination. Planta 151:189–197
Mesnard JM, Lebeurier G (1983) Arabidopsis thaliana cDNA bank. Research for genes coding for enzymes involved in pyrimidine metabolism. Arb Inf Serv 20:79–87
Negrutiu I, Beeftink F, Jacobs M (1975) Arabidopsis thaliana as a model system in somatic cell genetics I: cell and tissue culture. Plant Sci Lett 5:293–304
Negrutiu I, Jacobs M (1977) Arabidopsis as a model system in somatic cell genetics II: cell suspension cultures. Plant Sci Lett 8:7–15
Nokin P, Burny A, Cleuter Y, Huez G, Marbaix G, Chantrenne H (1975) Isolation and characterization of hichly purified globin mRNA from anaemic rabbit spleen. Eur J Biochem 53:83–90
O'Farrell P (1975) High resolution two dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
Pelham HRB, Jackson RJ (1976) An efficient mRNA dependent translation system from reticulocyte lysates. Eur J Biochem 67:247–256
Redei GP (1974) Arabidopsis thaliana. In: King RC (ed) Handbook of genetics, vol 2. Plenum Press, New York, p 151
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Röbbelen G (1975) The Laibach standard collections of natural races. Arab Inf Serv 2:36–47
Sachs MM, Freeling M (1978) Selective synthesis of alcohol dehydrogenase during anaerobic treatment of maize. Mol Gen Genet 161:111–115
Sachs MM, Freeling M, Okimoto R (1980) The anaerobic proteins of maize. Cell 20:761–767
Schwartz D, Osterman J (1976) A pollen selection system for alcohol dehydrogenase — negative mutants in plants. Genetics 83:63–65
Shimamoto K, King PJ (1983) Adh1-O: A selectable marker in Zea mays cell culture. Mol Gen Genet 191:271–275
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Taneja SR, Sachar RC (1976) Enzyme synthesis by conserved messengers in germinating wheat embryos. Phytochemistry 15:1589–1594
Thomas DS, Murashige T (1979a) Volatile emissions of plant tissue cultures. I. Identification of the major components. In Vitro 15:654–658
Thomas DS, Murashige T (1979b) Volatile emissions of plant tissue cultures. II. Effects of the auxin 2,4-D on production of volatiles in callus cultures. In Vitro 15:659–663
Verma DPS, Gordon AM, Byrne H, Ewings D (1975) Regulation and in vitro translation of messenger ribonucleic acid for cellulase from auxin-treated pea epicotyls. J Biol Chem 250:1019–1026
Werner M, Crabeel M, Glansdorff N, Pierard A (1982) Attempts at cloning Arabidopsis thaliana genes by transformation of yeast auxotrophic mutants. Arab Inf Serv 19:1–10
Zurfluh LL, Guilfoyle TJ (1982) Auxin-induced changes in the population of translatable messenger RNA in elongating sections of soybean hypocotyl. Plant Physiol 69:332–340
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Communicated by J. Schell
Dedicated to professor Georg Melchers to celebrate his 50-year association with the journal
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Dolferus, R., Marbaix, G. & Jacobs, M. Alcohol dehydrogenase in Arabidopsis: analysis of the induction phenomenon in plantlets and tissue cultures. Molec Gen Genet 199, 256–264 (1985). https://doi.org/10.1007/BF00330267
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DOI: https://doi.org/10.1007/BF00330267