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Differential expression of conglycinin α′ and β subunit genes in transgenic plants

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Abstract

β-conglycinin or 7S protein, one of the two major storage proteins of soybean (Glycine max), is comprised of three subunits, α′, α and β. Genomic clones encoding the α′ and β subunit genes were ligated together in intermediate plasmids, either in direct orientation (← ←) or in divergent orientation (← →). In the latter instance the transcriptional promoters of the two genes were approximately 2.0 kb apart. The DNAs were introduced into leaf disks of Petunia hybrida on a disarmed Ti plasmid in Agrobacterium tumefaciens and transgenic plants were regenerated. Expression of the introduced genes, limited to specific stages in developing seeds, was analyzed by quantitating the accumulated mRNA and protein, and compared with expression in transgenic plants that contained either gene alone. Development time course studies showed that the α′ subunit mRNA and protein were first detected at 10 days after pollination while accumulation of the β subunit protein ensued at day 14 regardless of the orientation of the two genes. The temporally regulated expression of these two genes in transgenic plants mimics their temporal regulation during soybean embryo development. In transgenic plants that contain the α′ and β genes in divergent orientation, expression of the β gene was enhanced several-fold, while the expression of the α′ gene was slightly decreased. These results suggest that (i) sequences of the α′ gene enhance the level of expression of a nearby storage protein gene and that (ii) the signal(s) operating in the temporal regulation of seed storage protein genes is (are) common to soybean and petunia, and are different from those that govern the final amounts of mRNA and protein.

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References

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

    Google Scholar 

  2. Bennett MD, Smith JB, Heslop-Harrison JS: Nuclear DNA amounts in angiosperms. Proc R Soc London Ser B 216: 179–199 (1982).

    Google Scholar 

  3. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).

    Google Scholar 

  4. Bray EA, Beachy RN: Regulation by ABA of β-conglycinin expression in cultured developing soybean cotyledons. Plant Physiol 79: 746–750 (1985).

    Google Scholar 

  5. Bray EA, Naito S, Pan N-S, Anderson E, Dubé P, Beachy RN: Expression of the β-subunit of β-conglycinin in seeds of transgenic plants. Planta 172: 364–370 (1987).

    Google Scholar 

  6. Chen Z-L, Schuler MA, Beachy RN: Functional analysis of regulatory elements in a plant embryo-specific gene. Proc Natl Acad Sci USA 83: 8560–8564 (1986).

    Google Scholar 

  7. Chen Z-L, Pan N-S, Beachy RN: A DNA sequence element that confers seed-specific enhancement to a constitutive promoter. EMBO J 7: 297–302 (1988).

    Google Scholar 

  8. Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).

    Google Scholar 

  9. Deom CM, Oliver MJ, Beachy RN. The 30-kilodalton gene product of tobacco mosaic virus potentiates virus movement. Science 237: 389–394 (1987).

    Google Scholar 

  10. Derbyshire E, Wright DJ, Boulter D: Legumin and vicilin, storage proteins of legume seeds. Phytochemistry (Oxf.) 15: 3–24 (1976).

    Google Scholar 

  11. Doyle JJ, Schuler MA, Godette WD, Zenger V, Beachy RN, Slightom JL: The glycosylated seed storage proteins of Glycine max and Phaseolus vulgaris. J Biol Chem 261: 9228–9238 (1986).

    Google Scholar 

  12. Fluhr R, Kuhlemeier C, Nagy F, Chua N-H: Organ-specific and light-induced expression of plant genes. Science 232: 1106–1112 (1986).

    Google Scholar 

  13. Fraley RT, Rogers SG, Horsch RB, Eichholtz DA, Flick JS, Fink CL, Hoffmann NL, Sanders PR: The SEV system: a new disarmed Ti-plasmid vector system for plant transformation. Biotechnology 3: 629–635 (1985).

    Google Scholar 

  14. Gayler KR, Sykes GE: β-conglycinins in developing soybean seeds. Plant Physiol 67: 958–961 (1981).

    Google Scholar 

  15. Gayler KR: Effects of nutritional stress on the storage proteins of soybeans. Plant Physiol 78: 582–585 (1985).

    Google Scholar 

  16. Goldberg RB, Hoscheck G, Ditta GS, Breidenbach RW: Developmental regulation of cloned superabundant embryo mRNAs in soybean. Dev Biol 83: 218–231 (1981).

    Google Scholar 

  17. Holowach LP, Thompson JF, Madison JT: Effects of exogenous methionine on storage protein composition of soybean cotyledons cultured in vitro. Plant Physiol 74: 576–583 (1984).

    Google Scholar 

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

    Google Scholar 

  19. Khoury G, Gruss P: Enhancer elements. Cell 33: 313–314 (1983).

    Google Scholar 

  20. Ladin BF, Tierney ML, Meinke DW, Hosangadi P, Veith M, Beachy RN: Developmental regulation of β-conglycinin in soybean axes and cotyledons. Plant Physiol 84: 35–41 (1987).

    Google Scholar 

  21. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  22. Meinke DW, Chen J, Beachy RN: Expression of storage-protein genes during soybean seed development. Planta 153: 130–139 (1981).

    Google Scholar 

  23. Nagy F, Morelli G, Fraley RT, Rogers SG, Chua N-H: Photoregulated expression of a pea rbcS gene in leaves of transgenic plants. EMBO J 4: 3063–3068 (1985).

    Google Scholar 

  24. Okamura JK, Jofuku KD, Goldberg RB: Soybean seed lectin gene and flanking nonseed protein genes are developmentally regulated in transformed tobacco plants. Proc Natl Acad Sci USA 83: 8240–8244 (1986).

    Google Scholar 

  25. Powell Abel P, Nelson RS, De B, Hoffmann N, Rogers SG, Fraley RT, Beachy RN: Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232: 738–743 (1986).

    Google Scholar 

  26. Rogers SG, Bisaro DM, Horsch RB, Fraley RT, Hoffmann NL, Brand L, Elmer JS, Lloyd AM: Tomato golden mosaic virus: A component DNA replicates autonomously in transgenic plants. Cell 45: 593–600 (1986).

    Google Scholar 

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

    Google Scholar 

  28. Schuler MA, Schmitt ES, Beachy RN: Closely related families of genes code for the α and α′ subunits of the soybean 7S storage protein complex. Nucl Acids Res 10: 8225–8244 (1985).

    Google Scholar 

  29. Schuttuck-Eidens DM, Beachy RN. 1985. Degradation of β-conglycinin in early stages of soybean embryogenesis. Plant Physiol 78: 895–898 (1985).

    Google Scholar 

  30. Tierney ML, Bray EA, Allen RD, Slightom JL, Drong RF, Yu Ma, Beachy RN: Isolation and characterization of a genomic clone encoding the β-subunit of β-conglycinin. Planta 172: 356–363 (1987).

    Google Scholar 

  31. Tumer NE, O'Connell KM, Nelson RS, Sanders PR, Beachy RN: Expression of alfalfa mosaic virus coat protein gene confers cross-protection in transgenic tobacco and tomato plants. EMBO J 6: 1181–1188 (1987).

    Google Scholar 

  32. Walling L, Drews GN, Goldberg RB: Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels. Proc Natl Acad Sci USA 83: 2123–2127 (1986).

    Google Scholar 

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Naito, S., Dubé, P.H. & Beachy, R.N. Differential expression of conglycinin α′ and β subunit genes in transgenic plants. Plant Mol Biol 11, 109–123 (1988). https://doi.org/10.1007/BF00015664

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  • DOI: https://doi.org/10.1007/BF00015664

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