Molecular and General Genetics MGG

, Volume 251, Issue 3, pp 267–280 | Cite as

Identification and isolation of theFEEBLY gene from tomato by transposon tagging

  • Erik A. van der Biezen
  • Bas F. Brandwagt
  • Wessel van Leeuwen
  • H. John J. Nijkamp
  • Jacques Hille
Original Paper
Received:
Accepted:

Abstract

TheAc/Ds transposon system from maize was used for insertional mutagenesis in tomato. Marker genes were employed for the selection of plants carrying a total of 471 uniqueDs elements. Three mutants were obtained withDs insertions closely linked to recessive mutations:feebly (fb), yellow jim (yj) anddopey (dp). Thefb seedlings produced high anthocyanin levels, developed into small fragile plants, and were insensitive to the herbicide phosphinothricin. Theyj plants had yellow leaves as a result of reduced levels of chlorophyll. Thedp mutants completely or partially lacked inflorescences. Thefb andyj loci were genetically linked to theDs donor site on chromosome 3. Reactivation of theDs element in thefb mutants by crosses with anAc-containing line resulted in restoration of the wild-type phenotypes. Plant DNA fragments flanking both sides of theDs element in thefb mutant were isolated by the inverse polymerase chain reaction. Molecular analysis showed that phenotypic reversions offb were correlated with excisions ofDs. DNA sequence analysis ofFb reversion alleles showed the characteristicDs footprints. Northern and cDNA sequence analysis indicated that transcription of theFEEBLY (FB) gene was impeded by the insertion ofDs in an intron. Comparison of the predicted amino acid sequence of the FB protein with other database sequences indicated thatFB is a novel gene.

Key words

Transposon tagging Activator/Dissociation Lycopersicon esculentum Mutants 
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aarts MGM, Dirkse WG, Stiekema WJ, Pereira A (1993) Transposon tagging of a male sterility gene in Arabidopsis. Nature 363:715–717PubMedGoogle Scholar
  2. Altmann T, Felix G, Jessop A, Kauschmann A, Uwer U, Peña-Cortés H, Willmitzer L (1995)Ac/Ds transposon mutagenesis inArabidopsis thaliana: mutant spectrum and frequency ofDs insertion mutants. Mol Gen Genet 247:646–652PubMedGoogle Scholar
  3. Bancroft I, Jones JDG, Dean C (1993) Heterologous transposon tagging of theDRL1 locus in Arabidopsis. Plant Cell 5:631–638PubMedGoogle Scholar
  4. Carroll BJ, Klimyuk VI, Thomas CM, Bishop GJ, Harrison K, Scofield SR, Jones JDG (1995) Germinal transpositions of the maize elementDissociation from T-DNA loci in tomato. Genetics 139:407–420PubMedGoogle Scholar
  5. Chuck G, Robbins T, Nijjar C, Ralston E, Courtney-Gutterson N, Dooner HK (1993) Tagging and cloning of a petunia flower color gene with the maize transposable elementActivator. Plant Cell 5:371–378PubMedGoogle Scholar
  6. De Block M, Botterman J, Vandewiele M, Donckx J, Thoen C, Gossele V, RaoMovva N, Thompson C, Van Montagu M, Leemans J (1987) Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J 6:2513–2518Google Scholar
  7. Fedoroff NV, Furtek DB, Nelson OE (1984) Cloning of thebronze locus in maize by a simple and generalizable procedure using the transposable controlling elementActivator (Ac). Proc Natl Acad Sci USA 81:3825–3829Google Scholar
  8. Feldmann KA (1991) T-DNA insertion mutagenesis in Arabidopsis: mutational spectrum. Plant J 1:71–82Google Scholar
  9. Gierl A, Saedler H (1992) Plant transposable elements and transposon tagging. Plant Mol Biol 19:39–49PubMedGoogle Scholar
  10. Haring MA, Rommens CMT, Nijkamp HJJ, Hille J (1991) The use of transgenic plants to understand transposition mechanisms and to develop transposon tagging strategies. Plant Mol Biol 16:449–461PubMedGoogle Scholar
  11. James DW, Lim E, Keller J, Plooy I, Ralston E, Dooner HK (1995) Directed tagging of the ArabidopsisFATTY-ACID ELONGATION 1 (FAE1) gene with the maize transposonActivator. Plant Cell 7:309–319PubMedGoogle Scholar
  12. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions:β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907PubMedGoogle Scholar
  13. Jones DA, Thomas CM, Hammond-Kosack KE, Balint-Kurti PJ, Jones JDG (1994) Isolation of the tomatoCf-9 gene for resistance toCladosporium fulvum by transposon tagging. Science 266:789–793PubMedGoogle Scholar
  14. Jones JDG, Jones DA, Bishop GJ, Harrison K, Carroll BJ, Scofield SR (1993) Use of the maize transposonsActivator andDissociation to show that phosphinothricin and spectinomycin resistance genes act non-cell-autonomously in tobacco and tomato seedlings. Transgenic Res 2:63–78Google Scholar
  15. Knapp S, Larondelle Y, Rossberg M, Furtek D, Theres K (1994) Transgenic tomato lines containingDs elements at defined genomic positions as tools for targeted transposon tagging. Mol Gen Genet 243:666–673PubMedGoogle Scholar
  16. Kunze R, Stochaj U, Laufs J, Starlinger P (1987) Transcription of transposable elementActivator (Ac) ofZea mays L. EMBO J 6:1555–1563Google Scholar
  17. Lawrence GJ, Finnegan EJ, Ayliffe MA, Ellis JG (1995) TheL6 gene for flax rust resistance is related to the Arabidopsis bacterial resistance geneRPS2 and the tobacco viral resistance geneN. Plant Cell 7:1195–1206PubMedGoogle Scholar
  18. Long D, Swinburne J, Martin M, Wilson K, Sundberg E, Lee K, Coupland G (1993) The maize transposable element systemAc/Ds as a mutagen in Arabidopsis: identification of an albino mutation induced byDs insertion. Proc Natl Acad Sci USA 90:370–374Google Scholar
  19. Martin C, Carpenter R, Sommer H, Saedler H, Coen ES (1985) Molecular analysis of instability of flower pigmentation inAntirrhinum majus following isolation of thepallida locus by transposon tagging. EMBO J 4:1625–1630Google Scholar
  20. Overduin B, Hogenhout SA, Van der Biezen EA, Haring MA, Nijkamp HJJ, Hille J (1993) TheAsc locus for resistance to Alternaria stem canker in tomato does not encode the enzyme aspartate carbamoyltransferase. Mol Gen Genet 240:43–48PubMedGoogle Scholar
  21. Rabino I, Mancinelli AL (1986) Light, temperature and anthocyanin production. Plant Physiol 81:922–924Google Scholar
  22. Salisbury FB, Ross CW (1992) Plant physiology, 4th edn. Wadsworth, Belmont, Calif., USAGoogle Scholar
  23. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  24. Scofield SR, Harrison K, Nurrish SJ, Jones JDG (1992) Promoter fusions to theActivator transposase gene cause distinct patterns ofDissociation excision in tobacco cotyledons. Plant Cell 4:573–582PubMedGoogle Scholar
  25. Souer E, Quattrocchio F, De Vetten N, Mol J, Koes R (1995) A general method to isolate genes tagged by a high copy number transposable element. Plant J 7:677–685PubMedGoogle Scholar
  26. Stam P (1993) Construction of integrated genetic linkage maps by means of a new computer package: JoinMap. Plant J 3:739–744Google Scholar
  27. Stevens MA, Rick CM (1986) Genetics and breeding. In: Atherton JG, Rudich J (eds) The tomato crop, a scientific basis for crop improvement. Chapman and Hall, New York, pp 35–100Google Scholar
  28. Sutton WD, Gerlach WL, Schwartz D, Peacock WJ (1984) Molecular analysis ofDs controlling element mutations at theAdh1 locus of maize. Science 223:1265–1268Google Scholar
  29. Thomas CM, Jones DA, English JJ, Carroll BJ, Bennetzen JF, Harrison K, Burbidge A, Bishop GJ, Jones JDG (1994) Analysis of the chromosomal distribution of transposon-carrying T-DNAs in tomato using the inverse polymerase chain reaction. Mol Gen Genet 242:573–585PubMedGoogle Scholar
  30. Van der Biezen EA, Overduin B, Nijkamp HJJ, Hille J (1994a) Integrated genetic map of tomato chromosome 3. Tomato Genet Coop Rep 44:8–10Google Scholar
  31. Van der Biezen EA, Van Haaren MJJ, Overduin B, Nijkamp HJJ, Hille J (1994b) Heterologous transposon tagging as a tool for the isolation of plant genes. In: Gelvin SB, Schilperoort RA (eds) Plant molecular biology manual. Kluwer, Dordrecht, pp K2:1–16Google Scholar
  32. Van der Biezen EA, Glagotskaya T, Overduin B, Nijkamp HJJ, Hille J (1995) Inheritance and genetic mapping of resistance toAlternaria alternata f. sp.lycopersici fromLycopersicon pennellii. Mol Gen Genet 247:453–461PubMedGoogle Scholar
  33. Walbot V (1992) Strategies for mutagenesis and gene cloning using transposon tagging and T-DNA insertional mutagenesis. Annu Rev Plant Physiol Plant Mol Biol 43:49–82Google Scholar
  34. Watterson JC (1985) Tomato diseases. A practical guide for seedsmen, growers and agricultural advisors. Petoseed, Calif., USAGoogle Scholar
  35. Whitham S, Dinesh-Kumar SP, Choi D, Hehl R, Corr C, Baker B (1994) The product of the tobacco mosaic virus resistance geneN: similarity to Toll and Interleukin-1 receptor. Cell 78:1101–1115PubMedGoogle Scholar
  36. Yoder JI (1990) Rapid proliferation of the maize transposable elementActivator in transgenic tomato. Plant Cell 2:723–730PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Erik A. van der Biezen
    • 1
  • Bas F. Brandwagt
    • 1
  • Wessel van Leeuwen
    • 1
  • H. John J. Nijkamp
    • 1
  • Jacques Hille
    • 1
  1. 1.Department of Genetics, Institute for Molecular Biological Sciences (IMBW), BioCentrum AmsterdamVrije UniversiteitAmsterdamThe Netherlands

Personalised recommendations