Summary
We have characterized a gene, 9612, that is expressed predominantly in the styles of tomato pistils according to a tightly regulated temporal program. 9612 RNA levels were maximal in mature pistils from flowers at anthesis, with transcripts undetectable in pistils from flowers collected 5–7 days prior to anthesis. In situ localization of mRNA in tissue sections showed that expression of the gene is confined in the pistil to the outer five cell layers of the strands of transmitting tissue within the upper two-thirds of the style. The maximal levels of 9612 RNA detected in anthers and vegetative organs were more than 50-fold and 250-fold lower than the level in pistils, respectively. A homolog to the 9612 gene was detected in tobacco and was also found to be expressed predominantly in the style. The ability of the 5′ flanking region of the tomato gene to appropriately regulate expression of a heterologous coding sequence was examined in transformed tomato and tobacco plants. In contrast to results with previously described regulated genes, the 9612 promoter functions correctly in the pistils of tomato plants, but fails to direct correct expression in tobacco plants. The sequence of the 9612 cDNA includes an open reading frame encoding a polypeptide of 404 amino acids with a highly hydrophobic amino-terminal region that may represent a signal peptide.
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Anderson MA, Cornish EC, Mau S-L, Williams EG, Hoggart R, Atkinson A, Bonig I, Grego B, Simpson R, Roche PJ, Haley JD, Penschow JD, Niall HD, Tregear GW, Coghlan JP, Crawford RJ, Clarke AE (1986) Cloning of a cDNA for a stylar glycoprotein associated with expression of self-incompatibility in Nicotiana alata. Nature 321:38–44
Benfey PN, Chua N-H (1989) Regulated genes in transgenic plants. Science 244:174–181
Breathnach R, Chambon P (1981) Organization and expression of eukaryotic split genes coding for proteins. Annu Rev Biochem 50:349–383
Buratowski S, Hahn S, Sharp PA, Guarente L (1988) Function of a yeast TATA element-binding protein in a mammalian transcription system. Nature 334:37–42
Chodosh LA, Baldwin AS, Carthew RW, Sharp PA (1988) Human CCAAT-binding proteins have heterologous subunits. Cell 53:11–24
Cresti M, van Went JL, Pacini E, Willemse MTM (1976) Ultrastructure of transmitting tissue of Lycopersicon peruvianum style: Development and histochemistry. Planta 132:305–312
Crouse GF, Frischauf A, Lehrach H (1983) An integrated and simplified approach to cloning into plasmids and single-stranded phage. Methods Enzymol 101:78–89
Davis RW, Botstein D, Roth JR (1980) Advanced Bacterial Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Day ML, Schwartz D, Wiegand RC, Stockman PT, Brunnert SR, Tolunay HE, Currie MG, Standaert DG, Needleman P (1987) Ventricular atriopeptin: Unmasking of messenger RNA and peptide synthesis by hypertrophy or dexamethasone. Hypertension 9:485–491
Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
Ebert PR, Anderson MA, Bernatzky R, Altschuler M, Clarke AE (1989) Genetic polymorphism of self-incompatibility in flowering plants. Cell 56:255–262
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Fluhr R, Chua N-H (1986) Developmental regulation of two genes encoding ribulose-bisphosphate carboxylase small subunit in pea and transgenic petunia plants: Phytochrome response and blue-light induction. Proc Natl Acad Sci USA 83:2358–2362
Fraley RT, Rogers SG, Horsch RB, Eichholtz DA, Flick JS, Fink CL, Hoffmann NL, Sanders PR (1985) The SEV system: A new disarmed Ti plasmid vector system for plant transformation. Biotechnology 3:629–635
Gasser CS, Smith AG, Budelier KA, Hinchee MA, McCormick S, Horsch RB, Shah DM, Fraley RT (1988) Isolation of differentially expressed genes from tomato flowers. In: Verma DPS, Goldberg RB (eds) Plant gene research, vol 5: Temporal and spatial regulation of plant genes. Springer-Verlag, Wien, New York, pp 83–96
Gasser CS, Budelier KA, Smith AG, Shah DM, Fraley RT (1989) Isolation of tissue-specific cDNAs from tomato pistils. Plant Cell 1:15–24
Goldberg RB (1988) Plants: Novel developmental processes. Science 240:1460–1467
Hanson DD, Hamilton DA, Travis JL, Bashe DM, Mascarenhas JP (1989) Characterization of a pollen-specific cDNA clone from Zea mays and its expression. Plant Cell 1:173–179
Heslop-Harrison J (1983) Self-incompatibility: phenomenology and physiology. Proc R Soc Lond [Biol] 218:371–395
Hinchee MAW, Connor-Ward DV, Newell CA, McDonnell RE, Sato SJ, Gasser CS, Fischhoff DA, Re DB, Fraley RT, Horsch RB (1988) Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer. Biotechnology 6:915–922
Horsch RB, Fry JE, Hoffmann NL, Wallroth M, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring cloned 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
Kamalay JC, Goldberg RB (1980) Regulation of structural gene expression in tobacco. Cell 19:935–946
Kislev N, Rubenstein I (1980) Utility of ethidium bromide in the extraction from whole plants of high molecular weight maize DNA. Plant Physiol 66:1140–1143
Kunkel TA (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132
Lalonde BA, Nasrallah ME, Dwyer KG, Chen C-H, Barlow B, Nasrallah JB (1989) A highly conserved Brassica gene with homology to the S-locus-specific glycoprotein structural gene. Plant Cell 1:249–258
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Mau S-L, Williams EG, Atkinson A, Anderson MA, Cornish EC, Grego B, Simpson RJ, Kheyr-Pour A, Clarke AE (1986) Style proteins of a wild tomato (Lycopersicon peruvianum) associated with expression of self-incompatibility. Planta 169:184–191
McCormick S, Niedermeyer J, Fry J, Barnason A, Horsch R, Fraley R (1986) Leaf disc transformation of cultivated tomato (L. esculentum) using Agrobacterium tumefaciens. Plant Cell Rep 5:81–84
McCormick S, Smith A, Gasser C, Sachs K, Hinchee M, Horsch R, Fraley R (1987) Identification of genes specifically expressed in reproductive organs of tomato. In: Niven DJ, Jones RA (eds) Tomato Biotechnology. Alan R Liss, New York, pp 255–265
Nasrallah JB, Kao T-H, Goldberg ML, Nasrallah ME (1985) A cDNA clone encoding an S-locus specific glycoprotein from Brassica oleracea. Nature 318:263–267
Sanders LC, Lord EM (1989) Directed movement of latex particles in the gynoecia of three species of flowering plants. Science 243:1606–1608
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schernthaner JP, Matzke MA, Matzke AJM (1988) Endospermspecific activity of a zein gene promoter in transgenic tobacco plants. EMBO J 7:1249–1255
Smith AG, Hinchee MA, Horsch RB (1987) Cell and tissue specific expression localized by in situ RNA hybridization in floral tissues. Plant Mol Biol Rep 5:237–241
Smith AG, Gasser CS, Budelier KA, Fraley RT (1990) Isolation of cDNAs preferentially expressed in tomato stamens. Mol Gen Genet 222:9–16
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Stinson JR, Eisenberg AJ, Willing RP, Pe ME, Hanson DD, Mascarenhas JP (1987) Genes expressed in the male gametophyte of flowering plants and their isolation. Plant Physiol 83:442–447
Ursin VM, Yamaguchi J, McCormick S (1989) Gametophytic and sporophytic expression of anther-specific genes in developing tomato anthers. Plant Cell 1:727–736
Vieira J, Messing J (1987) Production of single stranded DNA. Methods Enzymol 153:3–11
Voelker T, Sturm A, Chrispeels MJ (1987) Differences in expression between two seed lectin alleles obtained from normal and lectin-deficient beans are maintained in transgenic tobacco. EMBO J 6:3571–3577
von Heijne G (1986) A new method for predicting signal peptide cleavage sites. Nucleic Acids Res 14:4683–4690
von Heijne G, Abrahmsén L (1989) Species-specific variation in signal peptide design. FEBS Lett 244:439–446
Wagh PV, Bahl OP (1981) Sugar residues on proteins. CRC Crit Rev Biochem 10:307–377
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
Young RA, Davis RW (1983) Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci USA 80:1194–1198
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Communicated by E. Meyerowitz
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Budelier, K.A., Smith, A.G. & Gasser, C.S. Regulation of a stylar transmitting tissue-specific gene in wild-type and transgenic tomato and tobacco. Molec. Gen. Genet. 224, 183–192 (1990). https://doi.org/10.1007/BF00271551
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DOI: https://doi.org/10.1007/BF00271551