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Isolation and characterization of a light-inducible, organ-specific gene from potato and analysis of its expression after tagging and transfer into tobacco and potato shoots

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Summary

The isolation and analysis of several cDNAs and of one genomic clone encoding ST-LS1, a single-copy gene fromSolanum tuberosum with leaf/stem-specific, light-inducible expression, is described. The structure of the gene was determined by sequencing several overlapping partial cDNA clones as well as the genomic clone and by determining the transcription start site by RNA protection experiments. A “tagged” derivative of the gene, obtained by exon modification, was reintroduced into potato and into tobacco shoots usingAgrobacterium/Ti-plasmid vector systems. The modified gene was expressed in both tobacco and potato shoots giving rise to an RNA of approximately 1,200 nucleotides which exceeded the length of the RNA made from the endogenous gene by the expected size of the “tag” (470 nucleotides). The level of expression of the modified gene varied substantially between independent transformants. A high proportion of the transformants (20%–40%) synthesized as much RNA from the added gene as from the resident gene. Expression of the transferred gene was light induced. Qualitatively and quantitatively the expression of the introduced gene was similar in a homologous (potato) and in a heterologous, but related, cellular background (tobacco).

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References

  • Beachy RN, Chen ZL, Horsch RB, Rogers SG, Hoffmann NJ, Fraley RT (1985) Accumulation and assembly of soybean β-conglycinin in seeds of transformed petunia plants. EMBO J 4:3047–3053

    PubMed  CAS  Google Scholar 

  • Bevan M, Flavell RB, Chilton MD (1983) A chimaeric antibiotic resistance gene as a selectable marker for plant cell transformation. Nature 304:184–187

    Article  CAS  Google Scholar 

  • Broglie R, Coruzzi G, Fraley RT, Rogers SG, Horsch RB, Niedermeyer JG, Fink CL, Flick JS, Chua NH (1984) Light-regulated expression of a pea ribulose-1,5-bisphosphate carboxylase small subunit gene in transformed plant cells. Science 224:838–843

    PubMed  CAS  Google Scholar 

  • De Block M, Herrera-Estrella L, Van Montagu M, Schell J, Zambryski P (1984) Expression of foreign genes in regenerated plants and in their progeny. EMBO J 3:1681–1684

    PubMed  Google Scholar 

  • Dean C, Tamaki S, Dunsmuir P, Favreau M, Katayama C, Dooner H, Bedbrook J (1986) mRNA transcripts of several plant genes are polyadenylated at multiple sites in vivo. Nucleic Acids Res 14:2229–2240

    PubMed  CAS  Google Scholar 

  • Eckes P, Schell J, Willmitzer L (1985) Organ-specific expression of threee leaf/stem specific cDNA's from potato is regulated by light and correlated with chloroplast development. Mol Gen Genet 199:216–224

    Article  CAS  Google Scholar 

  • Fraley RT, Rogers SG, Horsch RB, Sanders PR, Flick JS, Adams SP, Bittner ML, Brand LA, Fink CL, Fry JS, Galluppi GR, Goldberg SB, Hoffmann NL, Woo SC (1983) Expression of bacterial genes in plant cells. Proc Natl Acad Sci USA 80:4803–4807

    Article  PubMed  CAS  Google Scholar 

  • Frischauf AM, Lehrach H, Poustka A, Murray N (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842

    PubMed  CAS  Google Scholar 

  • Gielen J, De Beuckeleer M, Seurinck J, Deboeck F, De Greve H, Lemmers M, Van Montagu M, Schell J (1984) The complete nucleotide sequence of the TL-DNA of theAgrobacterium tumefaciens plasmid pTiAch5. EMBO J 3:835–846

    PubMed  CAS  Google Scholar 

  • Gillies SD, Morrison SL, Oi YT, Tonegawa S (1983) A tissue specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell 33:717–728

    Article  PubMed  CAS  Google Scholar 

  • Gorman CM, Moffat CF, Howard BA (1982) Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2:1044–1051

    PubMed  CAS  Google Scholar 

  • Herrera-Estrella L, Depicker A, Van Montagu M, Schell J (1983a) Expression of chimaeric genes transferred into plant cells using a Ti-plasmid derived vector. Nature 303:209–213

    Article  CAS  Google Scholar 

  • Herrera-Estrella L, De Block M, Messens E, Hernalsteens JP, Van Montagu M, Schell J (1983b) Chimaeric genes as dominant selectable markers in plant cells. EMBO J 2:987–995

    PubMed  CAS  Google Scholar 

  • Herrera-Estrella L, Van den Broeck G, Maenhaut R, Van Montagu M, Schell J, Timko M, Cashmore A (1984) Light-inducible and chloroplast-associated expression of a chimaeric gene introduced intoNicotiana tabacum using a Ti-plasmid vector. Nature 310:115–120

    Article  PubMed  CAS  Google Scholar 

  • Horsch RB, Fry FE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    CAS  Google Scholar 

  • Jones J, Dunsmuir P, Bedbrook J (1985) High level expression of introduced chimaeric genes in regenerated transformed plants. EMBO J 4:2411–2418

    PubMed  CAS  Google Scholar 

  • Joos H, Inzew D, Caplan A, Sormann M, Van Montagu M, Schell J (1983) Genetic analysis of T-DNA transcripts in nopaline crown galls. Cell 32:1057–1067

    Article  PubMed  CAS  Google Scholar 

  • Kaulen H, Schell J, Kreuzaler F (1986) Light-induced expression of the chimeric chalcone synthase NPTII gene in tobacco cells. EMBO J 5:1–8

    PubMed  CAS  Google Scholar 

  • Lamppa G, Nagy F, Chua NH (1985) Light-regulated and organ-specific expression of a wheat Cab gene in transgenic tobacco. Nature 316:750–752

    Article  PubMed  CAS  Google Scholar 

  • Maniatis T, Fritsch E, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Press, New York, USA

    Google Scholar 

  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR (1984) Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage sp6 promoter. Nucleic Acids Res 12:7035–7056

    PubMed  CAS  Google Scholar 

  • Messing J, Geraghty D, Heidecker G, Hu NT, Kridl J, Rubenstein J (1983) Plant gene structure. In: Kosuge T, Meredith CP, Hollaender A (eds) Genetic engineering of plants — an agricultural perspective. Plenum Press, NY, pp 211–227

    Google Scholar 

  • Morelli G, Nagy F, Fraley RT, Rogers RG, Chua NH (1985) A short conserved sequence is involved in the light-inducibility of a gene encoding ribulose-1,5-bisphosphate carboxylase small subunits of pea. Nature 315:200–204

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Odell JT, Nagy F, Chua NH (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35 S promoter. Nature 313:810–812

    Article  PubMed  CAS  Google Scholar 

  • Rosahl S, Eckes P, Schell J, Willmitzer L (1986a) Organ-specific gene expression in potato: isolation and characterization of tuber-specific cDNA sequences. Mol Gen Genet 202:368–373

    Article  CAS  Google Scholar 

  • Rosahl S, Schmidt R, Schell J, Willmitzer L (1986b) Isolation and characterization of a gene fromSolanum tuberosum encoding patatin, the major storage protein of potato tubers. Mol Gen Genet 203:214–220

    Article  CAS  Google Scholar 

  • Sanchez-Serrano J, Schmidt R, Schell J, Willmitzer L (1986) Nucleotide sequence of proteinase inhibitor II encoding cDNA of potato and its mode of expression. Mol Gen Genet 203:15–20

    Article  CAS  Google Scholar 

  • Schoeffl F, Baumann G (1985) Thermo-induced transcripts of a soybean heat shock gene after transfer into sunflower using a Ti-plasmid vector. EMBO J 4:1119–1124

    CAS  Google Scholar 

  • Sengupta-Gopalan C, Reichert NA, Barker RF, Hall TC, Kemp JD (1985) Developmentally regulated expression of the bean phaseolin gene in tobacco seed. Proc Natl Acad Sci USA 82:3320–3324

    Article  PubMed  CAS  Google Scholar 

  • Shaw CH, Carter GH, Watson MD, Shaw CH (1984) A functional map of the nopaline synthase promoter. Nucleic Acids Res 12:7831–7846

    PubMed  CAS  Google Scholar 

  • Simpson J, Timko MP, Cashmore AR, Schell J, Van Montagu M, Herrera-Estrella L (1985) Light-inducible and tissue-specific expression of a chimaeric gene under control of the 5′-flanking sequence of a pea chlorophyll a/b binding protein gene. EMBO J 4:2723–2729

    PubMed  CAS  Google Scholar 

  • Spradling AC, Rubin GM (1983) The effect of chromosome position on the expression of theDrosophila xanthine dehydrogenase gene. Cell 34:47–57

    Article  PubMed  CAS  Google Scholar 

  • Van Haute E, Joos H, Maes M, Warren G, Van Montagu M, Schell J (1983) Intergenic transfer and exchange recombination of restriction fragments cloned in pBR322: a novel strategy for the reversed genetics of the Ti-plasmid ofAgrobacterium tumefaciens. EMBO J 2:411–418

    PubMed  Google Scholar 

  • Weiher H, Koenig M, Gruss P (1983) Multiple point mutations affect the SV40 enhancer. Science 219:626–631

    PubMed  CAS  Google Scholar 

  • Werr W, Frommer WB, Maas C, Starlinger P (1985) Structure of the sucrose synthase gene on chromosome 9 ofZea mays L. EMBO J 4:1373–1380

    PubMed  CAS  Google Scholar 

  • Willmitzer L, Dhaese P, Schreier P, Schmalenbach W, Van Montagu M, Schell J (1983) Size, location and polarity of T-DNA encoded transcripts in nopaline crown gall tumors, common transcripts in octopine and nopaline tumors, Cell 32:1045–1056

    Article  PubMed  CAS  Google Scholar 

  • Zambryski P, Joos H, Genetello C, Leemans J, Van Montagu M, Schell J (1983) Ti-plasmid vector for the introduction of DNA into plant cells without alteration of their normal regeneration capacity. EMBO J 2:2143–2150

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Max Planck-Institut für Züchtungsforschung, D-5000, Köln 30, Federal Republic of Germany

    Peter Eckes, Sabine Rosahl, Jeff Schell & Lothar Willmitzer

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  1. Peter Eckes
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  2. Sabine Rosahl
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  3. Jeff Schell
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  4. Lothar Willmitzer
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Communicated by P. Starlinger

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Eckes, P., Rosahl, S., Schell, J. et al. Isolation and characterization of a light-inducible, organ-specific gene from potato and analysis of its expression after tagging and transfer into tobacco and potato shoots. Molec. Gen. Genet. 205, 14–22 (1986). https://doi.org/10.1007/BF02428027

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  • DOI: https://doi.org/10.1007/BF02428027

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