Skip to main content
Log in

Genetic and molecular characterization of an Adh-1 null mutant in tomato

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Treatment of tomato seeds with ethyl methanesulphonate (EMS) followed by allyl alcohol selection of M2 seeds has led to the identification of one plant (B15-1) heterozygous for an alcohol dehydrogenase (Adh) null mutation. Genetic analysis and expression studies indicated that the mutation corresponded to the structural gene of the Adh-1 locus on chromosome 4. Homozygous Adh-1 null mutants lacked ADH-1 activity in both pollen and seeds. Using an antiserum directed against ADH from Arabidopsis thaliana, which crossreacts with ADH-1 and ADH-2 proteins from tomato, no ADH-1 protein was detected in seeds of the null mutant. Northern blot analysis showed that Adh-1 mRNA was synthesized at wild-type levels in immature seeds of the null mutant, but dropped to 25% in mature seeds. Expression of the Adh-2 gene on chromosome 6 was unaffected. The potential use of the Adh-1 null mutant in selecting rare transposon insertion mutations in a cross with “mutable” Adh-1 + tomato lines is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bicsak TA, Kann LR, Reiter A, Chase T (1982) Tomato alcohol dehydrogenase: purification and substrate specificity. Arch Biochem Biophys 216:605–615

    Google Scholar 

  • Bonas U, Sommer H, Harrison BJ, Saedler H (1984) The transposable element Tam 1 of Anthirrhinum majus is 17 kb long. Mol Gen Genet 194:138–143

    Google Scholar 

  • Chase T, Williams BG (1986) Characterization of complete cDNA for tomato Adh-2. Abstract Tomato Biotechnology Symposium, University of California, Davis, August 20–22, 1986

  • Chourey PS, Nelson OE (1976) The enzymatic deficiency conditioned by the shrunken-1 mutations in maize. Biochem Genet 14:1041–1055

    Google Scholar 

  • 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

    Google Scholar 

  • De Vries S, Hoge H, Bisseling T (1988) Isolation of total and polysomal RNA from plant tissues. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Döring HP, Freeling M, Hake S, Johns MA, Kunze P, Merckelbach A, Salamini F and Starlinger P (1984a) A Ds-mutation of the Adh-1 gene in Zea mays L. Mol Gen Genet 193:199–204

    Google Scholar 

  • Döring HP, Tillmann E, Starlinger P (1984b) DNA sequence of the maize transposable element Dissociation. Nature 307:127–130

    Google Scholar 

  • Fedoroff N, Wessler S, Shure M (1983) Isolation of the transposable maize controlling elements Ac and Ds. Cell 35:235–242

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radio-labeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–10

    Google Scholar 

  • Freeling M (1976) Intragenic recombination in maize: pollen analysis methods and the effect of the parental Adh-1 + isoalleles. Genetics 83:707–717

    Google Scholar 

  • Freeling M, Birchler JA (1981) Mutants and variants of the alcohol dehydrogenase-1 gene in maize. In: Setlow JK, Hollaender A (eds) Genetic engineering. Principles and methods (3). Plenum, New York London, pp 223–264

    Google Scholar 

  • Freeling M, Cheng DSK (1978) Radiation-induced alcohol dehydrogenase mutants in maize following allyl alcohol selection of pollen. Genet Res 31:107

    Google Scholar 

  • Geiser M, Week E, Döring HP, Courage-Tebbe U, Tillmann E, Starlinger P (1982) Genomic clones of wild type allele and a transposable element of the sucrose synthase gene of Zea mays L. EMBO J 1:1455–1460

    Google Scholar 

  • Gerlach WL, Pryor AJ, Dennis ES, Ferl RJ, Sachs MM, Peacock WJ (1982) DNA cloning and induction of the alcohol dehydrogenase gene (Adh-1) of maize. Proc Natl Acad Sci USA 79:2981–2985

    Google Scholar 

  • Grandbastien M, Spielman A, Caboche M (1989) Tntl, a mobile retroviral-like transposable element of tobacco isolated by plant cell genetics. Nature 337:376–380

    Google Scholar 

  • Jacobs M, Dolferus R, Van den Bosche D (1988) Isolation and biochemical analysis of ethyl methanesulfonate-induced alcohol dehydrogenase null mutants of Arabidopsis thaliana (L.) Heynh. Biochem Genet 26:105–122

    Google Scholar 

  • Khush GS, Rick CM (1967) Studies on the linkage map of chromosome 4 and on the transmission of induced deficiencies. Genetics 38:74–94

    Google Scholar 

  • Kuckuck H (1936) Über vier neue Serien multipler Allele bei Antirrhinum majus. Z Inductive Abstammungs- und Vererbungslehre 71:429–440

    Google Scholar 

  • Llewellyn DJ, Finnegan EJ, Ellis JG, Dennis ES, Peacock WJ (1987) Structure and expression of an alcohol dehydrogenase gene from Pisum sativum. J Mol Biol 195:115–123

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Martin P, Place AR, Pentz E, Sofer W (1985) UGA nonsense mutation in the alcohol dehydrogenase gene of Drosophila melanogaster. J Mol Biol 184:221–229

    Google Scholar 

  • Moffatt B, Somerville C (1988) Positive selection for male-sterile mutants of Arabidopsis lacking adenine phosphoribosyl transferase activity. Plant Physiol 86:1150–1154

    Google Scholar 

  • Ramanna MS, Hille J, Zabel P (1985) Chromosome breakage-fusion-bridge-cycle and phenotypic instability in isochromosome lines of tomato. Theor Appl Genet 71:145–152

    Google Scholar 

  • Rousselin P, Lipingle A, Faure J-D, Bitouin R, Caloche M (1990) Ethanol-resistant mutants of Nicotiana plumbaginifolia are deficient in the expression of pollen and seed alcohol dehydrogenase activity. Mol Gen Genet 222:409–415

    Google Scholar 

  • Schwartz D, Osterman J (1976) A pollen selection system for alcohol dehydrogenase-negative mutants in plants. Genetics 83:63–65

    Google Scholar 

  • Schwartz-Sommer Zs, Gierl A, Klösgen RB, Wienand U, Peterson PA, Saedler H (1984) The Spin (En) transposable element controls the excision of a 2-kb DNA insert at the wxm-8 allele of Zea mays. EMBO J 3:1021–1028

    Google Scholar 

  • Selander RK, Smith HH, Yang SY, Johnson WE, Gentry JB (1971) Biochemical polymorphisms and systematics in the genus Peromyscus. I. Variation in the old field mouse (Peromyscus polionotus). Univ Texas Publ 7103:49–90

    Google Scholar 

  • Sommer H, Saedler H (1986) Structure of the chalcone synthase gene of Antirrhinum majus. Mol Gen Genet 202:429–434

    Google Scholar 

  • Sprague GT, Brimhall B, Hixon RM (1943) Some effects of the waxy gene in corn on the properties of the endosperm starch. J Am Soc Agron 35:817–822

    Google Scholar 

  • Staraci LC, Thomas BR, Wainwright CA, Marsh E, Rejda JM, Steffens JC, Genez AL, Williams BG (1987) Stress adaptations in cultured tomato cells. In: Nevins DJ, Jones RA (eds) Tomato Biotechnology. Liss, New York, pp 99–108

    Google Scholar 

  • Sutton WD, Gerlach WL, Schwarz D, Peacock WJ (1984) Molecular analysis of Ds controlling element mutations at the Adh-1 locus of maize. Science 223:1265–1268

    Google Scholar 

  • Tanksley SD (1979) Linkage, chromosomal association, and expression of Adh and Pgm-2 in tomato. Biochem Genet 17:1159–1167

    Google Scholar 

  • Tanksley SD, Jones RA (1981) Effects of OZ stress on tomato alcohol dehydrogenase activity: description of a second ADH coding gene. Biochem Genet 19:397–409

    Google Scholar 

  • Weinand U, Sommer H, Schwarz Zs, Shepherd N, Saedler H, Kreuzaler F, Ragg H, Fautz E, Hahlbrock K, Harrison BJ, Peterson PA (1982) A general method to identify plant structural genes among genomic DNA clones using transposable element induced mutations. Mol Gen Genet 187:195–201

    Google Scholar 

  • Widholm JM, Kishinami I (1988) Allyl alcohol selection for lower alcohol dehydrogenase activity in Nicotiana plumbaginifolia cultured cells. Plant Physiol 86:266–269

    Google Scholar 

  • Xie Y, Wu R (1989) Molecular analysis of an alcohol dehydrogenase-encoding clone (adh2) from rice. Gene 87:185–191

    Google Scholar 

  • Zabel P, Moerman M, V. Straaten F, Goldbach R, V. Kammen A (1982) Antibodies against the genome-linked protein VPg of cowpea mosaic virus recognize a 60,000-dalton precursor polypeptide. J Virol 41:1083–1088

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H. Saedler

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wisman, E., Koornneef, M., Chase, T. et al. Genetic and molecular characterization of an Adh-1 null mutant in tomato. Mol Gen Genet 226, 120–128 (1991). https://doi.org/10.1007/BF00273595

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00273595

Key words

Navigation