Abstract
The maize transposable element, Activator (Ac), is being used to develop a transposon mutagenesis system in lettuce, Lactuca sativa. In this paper, we describe somatic and germinal transactivation of Ds by chimeric transposase genes in whole plants. Constructs containing either the Ds element or the Ac transposase open reading frame (ORF) were introduced into lettue. The Ds element was located between either the 35S or the Nos promoter and a chimeric spectinomycin resistance gene (which included a transit peptide), preventing expression of spectinomycin resistance. The genomic coding region of the Ac transposase was expressed from the 35S promoter. Crosses were made between 104 independent R1 plants containing Ds and three independent R1 plants expressing transposase. The excision of Ds in F1 progenies was monitored using a phenotypic assay on spectinomycin-containing medium. Green sectors in one-third of the F1 families indicated transactivation of Ds by the transposase at different developmental stages and at different frequencies in lettuce plants. Excision was confirmed using PCR and by Southern analysis. The lack of green sectors in the majority of F1 families suggests that the majority of T-DNA insertion sites are not conducive to excision. In subsequent experiments, the F1 plants containing both Ds and the transposase were grown to maturity and the F2 seeds screened on medium containing spectinomycin. Somatic excision was again observed in several F2 progeny; however, evidence for germinal excision was observed in only one F2 family.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Baker B, Schell J, Loerz H, Fedoroff N (1986) Transposition of the maize controlling element “Activator” in tobacco. Proc Natl Acad Sci USA 83:4844–4848
Bancroft I, Bhatt AM, Sjodin C, Scofield S, Jones JDG, Dean C (1992) Development of an efficient two-element transposon tagging system in Arabidopsis thaliana. Mol Gen Genet 233:449–461
Belzile F, Lassner MW, Tong Y, Khush R, Yoder JI (1989) Sexual transmission of transposed Activator elements in transgenic tomato. Genetics 123:181–189
Brink RA, Williams E (1973) Mutable R-Navajo alleles of cyclic origin in maize. Genetics 73:273–296
Dean C, Sjodin C, Page T, Jones J, Lister C (1992) Behavior of the maize transposable element Ac in Arabidopsis thaliana. Plant J 2:69–82
Dooner HK, Belachew A (1989) Transposition pattern of the maize element Ac from the bz-m2(Ac) allele. Genetics 122:447–457
Dooner HK, Keller J, Harper E, Ralston E (1991) Variable patterns of transposition of the maize element Activator in tobacco. Plant Cell 3:473–482
Fedoroff N (1989) Maize transposable elements. In: Berg DE, Howe MM (eds) Mobile DNA, American Society for Microbiology, Washington DC, pp 375–411
Hake S, Vollbrecht E, Freeling M (1989) Cloning Knotted, the dominant morphological mutant in maize using Ds2 as a transposon tag. EMBO J 8:15–22
Haring MA, Goa J, Volbeda T, Rommens C, Nijkamp J, Hille J (1989) A comparative study of Tam3 and Ac transposition in transgenic tobacco and petunia plants. Plant Mol Biol 13:189–201
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 16:449–461
Hehl R, Baker B (1989) Induced transposition of Ds by a stable Ac in crosses of transgenic tobacco plants. Mol Gen Genet 217:53–59
Hehl R, Baker B (1990) Properties of the maize transposable element Activator in transgenic tobacco plants: a versatile interspecies genetic tool. Plant Cell 2:709–921
Heinlein M, Starlinger P (1991) Variegation patterns caused by transposable element Ac. Maydica 36:309–316
Izawa T, Miyazaki C, Yamamoto M, Terada R, Iida S, Shimamoto K (1991) Introduction and transposition of the maize transposable element Ac in rice (Oryza sativa L.). Mol Gen Genet 227:391–396
Jones JDG, Carland FM, Maliga P, Dooner HK (1989) Visual detection of transposition of the maize element Activator (Ac) in tobacco seedlings. Science 244:204–207
Jones JDG, Carland F, Lim E, Ralston E, Dooner H (1990) Preferential transposition of the maize element Activator (Ac) to linked chromosomal locations in tobacco. Plant Cell 2:701–707
Jones JDG, Harper E, Carland F, Ralston E, Dooner HK (1991) Reversion and altered variegation of an SPT: Ac allele in tobacco. Maydica 36:329–335
Jones JDG, Shlumukov L, Carland F, English J, Scofield SR, Bishop GJ, Harrison K (1992) Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants. Transgenic Res 1:285–297
Knapp S, Coupland G, Uhrig H, Starlinger P, Salamini F (1988) Transposition of the maize transposable element Ac in Solanum tuberosum. Mol Gen Genet 213:285–290
Lassner MW, Palys JM, Yoder JI (1989) Genetic transactivation of Dissociation elements in transgenic tomato plants. Mol Gen Genet 218:25–32
Martin C, Carpenter R, Sommer H, Saedler H, Coen ES (1985) Molecular analysis of instability in flower pigmentation of Antirrhinum majus, following isolation of the pallida locus by transposon tagging. EMBO J 4:1625–1630
Masterson RV, Furtek DB, Grevelding C, Schell J (1989) A maize Ds transposable element containing a dihydrofelate reductase gene transposes in Nicotiana tabacum and Arabidopsis thaliana. Mol Gen Genet 219:461–466
McClintock B (1951) Chromosome organization and genic expression. Cold Spring Harbor Symp Quant Biol 16:13–47
McClintock B (1956) Mutation in maize. Carnegie Inst Washington Yearb 55:323–332
Michelmore RW, Marsh E, Seeley D, Landry BS (1987) Transformation of lettuce mediated by Agrobacterium tumefaciens. Plant Cell Rep 6:439–442
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325
Paz-Ares J, Wienand U, Peterson PA, Saedler H (1986) Molecular cloning of the c locus of Zea mays: a locus regulating the anthocyanin pathway. EMBO J 5:829–833
Schmidt R, Willmitzer L (1989) The maize autonomous element Activator (Ac) shows a minimal germinal excision frequency of 0.2%–0.5% in transgenic Arabidopsis thaliana plants. Mol Gen Genet 220:17–24
Scofield SR, Harrison K, Nurrish SJ, Jones JDG (1992) Promoter fusion to the Activator transposase gene confer distinct patterns of Dissociation excision in tobacco cotyledons. Plant Cell 4:573–582
Swinburne J, Balcells L, Scofield S, Jones J, Coupland G (1992) Elevated levels of Activator transposase mRNA are associated with high frequencies of Dissociation excision in Arabidopsis. Plant Cell 4:583–595
Theres N, Scheele T, Starlinger P (1987) Cloning of the Bz2 locus of Zea mays using the transposable element Ds as a gene tag. Mol Gen Genet 209:193–197
Van Sluys MA, Tempe J (1989) Behavior of the maize transposable element Ac in Daaucus carota. Mol Gen Genet 219:313–319
Van Sluys MA, Tempe J, Fedoroff N (1987) Studies on the introduction and mobility of the maize Activator element in Arabadopsis thaliana and Daucus carota. EMBO J 6:3881–3889
Yang C-H (1992) Behavior of the maize transposable element Ac in lettuce (Lactuca sativa). Ph D Thesis, University of California, Davis
Yang C-H, Ellis J, Michelmore RW (1993) Infrequent transposition of Ac in lettuce (Lactuca sativa). Plant Mol Biol, in press
Yoder JI, Belzile F, Alpert K, Palys J, Michelmore RW (1987) Mobilization of the maize transposable element Ac in tomato. In: Tomato Biotechnology, Plant Biology Series, Vol 4. Eds Nevins DJ and Jones RA. Alan Liss Inc., N.Y. pp 189–198
Zhou JH, Athcrly AG (1990) In situ detection of transposition of the maize controlling element (Ac) in transgenic soybean tissues. Plant Cell Rep 8:542–545
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
Rights and permissions
About this article
Cite this article
Yang, CH., Carroll, B., Scofield, S. et al. Transactivation of Ds elements in plants of lettuce (Lactuca sativa). Molec. Gen. Genet. 241, 389–398 (1993). https://doi.org/10.1007/BF00284692
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00284692