Abstract
Tissue-specific expression of the gene coding for trypsin inhibitor BTI-CMe in barley (Itr1) occurs during the first half of endosperm development. In transgenic tobacco, theItr1 promoter drives expression of the β-glucuronidase reporter gene not only in developing endosperm but also in embryo, cotyledons and the meristematic intercotyledonary zone of germinating seedlings. A promoter fragment extending 343 bp upstream of the translation initiation ATG codon was sufficient for full transgene expression, whereas, the proximal 83 bp segment of the promoter was inactive. Possible reasons for the differences in expression patterns are discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bevan M (1984) BinaryAgrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721
Cameron-Mills V, Brandt A (1988) A γ-hordein gene. Plant Mol Biol 11:449–461
Colot V, Robert LS, Kavanagh TA, Bevan MW, Thompson RD (1987) Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco. EMBO J 6:3559–3564
DeBlock M, Debrouwer D (1992) In situ enzyme histochemistry on plastic-embedded plant material. The development of an artefact-free β-glucuronidase assay. Plant J 2:261–266
Díaz I, Royo J, Carbonero P (1993) The promoter of barley trypsin-inhibitor BTI-CMe discriminates between wheat and barley endosperm protoplasts in transient expression assays. Plant Cell Rep 12:698–701
Doll H, Køie B, Eggum BO (1974) Induced high lysine mutants in barley. Radiation Bot 14:73–80
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Fernández JA, Moreno M, Carmona MJ, Castagnaro A, Garcia-Olmedo F (1993) The barley α-thionin promoter is rich in negative regulatory motifs and directs tissue-specific expression of a reporter gene in tobacco. Biochim Biophys Acta 1172:346–348
Forde BG, Heyworth A, Pywell J, Kreis M (1985) Nucleotide sequence of a β-1 hordein gene and the identification of possible upstream regulatory elements in endosperm storage protein genes from barley, wheat and maize. Nucleic Acids Res 13:7327–7339
Gallusci P, Salamini F, Thompson RD (1994) Differences in cell type-specific expression of the geneOpaque2 in maize and transgenic tobacco. Mol Gen Genet 244:391–400
Goldberg RB, Barker SJ, Pérez-Grau L (1989) Regulation of expression during plant embryogenesis. Cell 56:149–160
Hoekema A, Hirsch PR, Hooykass PJJ, Schilperoot RA (1983) A binary plant vector strategy based on the separation of vir- and T-region of theAgrobacterium tumefaciens Ti plasmid. Nature 303:179–180
Hopp HE, Rasmussen SK, Brandt A (1983) Organization and transcription of B1 hordein genes in high lysine mutants of barley. Carlsberg Res Commun 48:201–210
Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Kreis M, Shewry PR, Forde BG, Rahman S, Miffin BJ (1984) Molecular analysis of the effects of the mutantlys 3a gene on the expression ofHor loci in developing endosperms of barley (Hordeum vulgare). Cell 34:161–167
Kreis M, Williamson M, Buxton B, Pywell J, Heijgaard J, Svendsen I (1987) Primary structure and differential expression of β-amylase in normal and mutant barleys. Eur J Biochem 169:517–525
Lagrimini LM, Burkhart W, Moyer M, Rothstein S (1987) Molecular cloning of complementary DNA encoding the lignin forming peroxidase from tobacco: molecular analysis and tissue-specific expression. Proc Natl Acadl Sci USA 84:7542–7546
Lazaro A, Barber D, Salcedo G, Mendez E, Garcia-Olmedo F (1985) Differential effects of high lysine mutations on the accumulation of individual members of a group of proteins encoded by a disperse multigene family in the endosperm of barley (Hordeum vulgare L). Eur J Biochem 149:617–623
Lopes MA, Larkins BA (1993) Endosperm origin, development and function. Plant Cell 5:1383–1399
Maier UG, Grasser KD, Haass MM, Feix G (1990) Multiple proteins bind to the P2 promoter region of the zein pMS1 of maize. Mol Gen Genet 221:164–170
Marris C, Gallois P, Copley J, Kreis M (1989) The 5′ flanking region of a barley B hordein gene controls tissue and developmental specific CAT expression in tobacco plants. Plant Mol Biol 10:359–366
Matzke AJM, Stöger EM, Schrenthaner JP, Matzke MA (1990) Deletion analysis of a zein gene promoter in transgenic tobacco plants. Plant Mol Biol 14:323–332
Mikola J, Soulinna EM (1969) Purification and properties of a trypsin inhibitor from barley. Eur J Biochem 9:555–560
Motto M, DiFonzo N, Hartings H, Maddaloni M, Salamini F, Soave C, Thompson RD (1989) Regulatory genes affecting maize storage protein synthesis. In: Shewry PR (ed) Oxford surveys plant molecular and cell biology, vol 6. IRL Press, Oxford, 87–144
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Quayle T, Feix G (1992) Functional analysis of the — 300 region of maize zein genes. Mol Gen Genet 231:369–374
Reeves CD, Okita TW (1987) Analysis of α/β-type gliadin genes from diploid and hexaploid wheats. Gene 52:257–266
Roberts LS, Thompson RD, Flavell RB (1989) Tissue-specific expression of a wheat high molecular weight glutenin gene in transgenic tobacco. Plant Cell 1:569–578
Rodriguez-Palenzuela P, Royo J, Gómez L, Sánchez-Monge R, Salcedo G, Molina-Cano JL, Garcia-Olmedo F, Carbonero P (1989) The gene for trypsin inhibitor CMe is regulated in trans by thelys 3a locus in the endosperm of barley (Hordeum vulgare L). Mol Gen Genet 219:474–479
Salcedo G, Fra-Mon P, Molina-Cano JL, Aragoncillo C, Garcia-Olmedo F (1984) Genetics of CM-proteins (A-hordeins) in barley. Theor Appl Genet 68:53–59
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Schernthaner JP, Matzke MA, Matzke AJM (1988) Endosperm specific activity of a zein gene promoter in transgenic tobacco plants. EMBO J 7:1249–1255
Schmidt RJ, Burr FA, Aukerman MJ, Burr B (1990) Maize regulatory geneopaque2 encodes a protein with a “leuzine-zipper” motif that binds to zein DNA. Proc Natl Acad Sci USA 87:46–50
Schubert R, Panitz R, Manteuffel R, Nagy I, Wobus U, Bäumlein H (1994) Tissue-specific expression of an oat 12S seed globulin gene in developing tobacco seeds: differential mRNA and protein accumulation. Plant Mol Biol 26:203–210
Thompson GA, Boston RS, Lyznik LA, Hodges TK, Larkins BA (1990) Analysis of promoter activity from an α-zein gene 5′ flanking sequence in transient expression assays. Plant Mol Biol 15:755–764
Ueng P, Gallili G, Sapanara V, Goldsbrough PB, Dube P, Beachy RN (1988) Expression of a maize storage protein gene in petunia plants is not restricted to seeds. Plant Physiol 86:1281–1285
Author information
Authors and Affiliations
Additional information
Communicated by H. Saedler
These authors have contributed equally to this work
Rights and permissions
About this article
Cite this article
Diaz, I., Royo, J., O'Connor, A. et al. The promoter of the geneItr1 from barley confers a different tissue specificity in transgenic tobacco. Molec. Gen. Genet. 248, 592–598 (1995). https://doi.org/10.1007/BF02423455
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02423455