Skip to main content
Log in

Molecular and genetic analysis of an embryonic gene, DC 8, from Daucus carota L.

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

To understand the morphogenetic and physiological processes occurring during plant embryogenesis, we isolated cDNA clones homologous to genes preferentially expressed during somatic embryogenesis. One of these cDNA clones detected an embryo-specific mRNA species with a corresponding protein of 66 kDa. The expression pattern of the mRNA is similar between somatic and zygotic embryos of carrots. To characterize the gene encoding this mRNA, we isolated the corresponding genomic clones. Molecular analysis of the DNA from several haploid and diploid carrots showed that the mRNA was encoded by a single copy gene, named DC 8. DNA sequence analysis showed that the gene consisted of three exons and coded for a hydrophilic protein with a central region composed of 17 repeats. At the NH2-terminus no typical signal sequence was found. Immunocytochemical analysis localized the protein primarily in the vacuoles and protein bodies of zygotic embryos; the cytoplasm showed some antibody staining. The protein was also found in cell walls of endosperm tissue. The amount of DC 8 protein was too low for it to be categorized as a seed storage protein; its role in embryogenesis remains to be determined.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Aquadro CF, Desse SF, Bland MM, Langley CH, Laurie-Ahlberg CC (1986) Molecular population genetics of the alcohol dehydrogenase gene region of Drosophila melanogaster. Genetics 114:1165–1190

    Google Scholar 

  • Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acidcellulose. Proc Natl Acad Sci USA 69:1408–1412

    Google Scholar 

  • Baker J, Steele C, Dure III L (1988) Sequence and characterization of 6 Lea proteins and their genes from cotton. Plant Mol Biol, in press

  • Biggin MD, Gibson TJ, Hong GF (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80:3963–3965

    Google Scholar 

  • Borkird C, Choi JH, Jin ZH, Franz G, Hatzopoulos P, Chorneau R, Bonas U, Pelegri F, Sung ZR (1988) Developmental regulation of embryogenic genes in plants. Proc Natl Acad Sci USA 85:6399–6403

    Google Scholar 

  • Breathnach R Chambon P (1981) Organization and expression of eukaryotic split genes coding for proteins. Annu Rev Biochem 50:349–383

    Google Scholar 

  • Choi JH, Liu LS, Borkird C, Sung ZR (1987) Cloning of genes developmentally regulated during plant embryogenesis. Proc Natl Acad Sci USA 84:1906–1910

    Google Scholar 

  • Chou PY, Fasman GD (1974) Prediction of protein conformation. Biochemistry 13:222–245

    Google Scholar 

  • Church G, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995

    Google Scholar 

  • Dente L, Cesareni G, Cortese R (1983) pEMBL: a new family of single stranded plasmids. Nucleic Acids Res 11:1645–1655

    Google Scholar 

  • Foster AS, Gifford EM (1974) Comparative morphology of vascular plants. Freeman W. H. San Francisco

    Google Scholar 

  • Galau GA, Hughes DW (1987) Coordinate accumulation of homologous transcripts of seven cotton Lea gene families during embryogenesis and germination. Dev Biol 123:213–221

    Google Scholar 

  • Gamborg OL, Miller RA, Ohuama K (1968) Nutritional requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Google Scholar 

  • Gray D, Ward JA, Stechkel JRA (1984) Endosperm and embryo development in Daucus carota L. J Exp Bot 35:459–465

    Google Scholar 

  • Hultmark D, Klemenz R, Gehring W (1986) Translational and transcriptional control elements in the untranslated leader of the heatshock gene hsp22. Cell 44:429–438

    Google Scholar 

  • Huynh TV, Young RA, Davis RW (1985) Construction and screening of cDNA libraries in lambda gt10 and lambda gt11. In: Glover D (ed) DNA cloning: A practical guide. IRL Press, Oxford, England, pp 49–78

    Google Scholar 

  • Joshi CP (1987) An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucleic Acids Res 15:6643–6653

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Langley CH, Shrimpton AE, Yamazaki T, Miyashita N, Matsuo Y, Aquadro CF (1988) Naturally occurring variation in the restriction map of the amy region of Drosophila melanogaster. Genetics 119:619–629

    Google Scholar 

  • Lipman DJ, Pearson WR (1985) Rapid and sensitive protein similarity searches. Science 227:1435–1441

    Google Scholar 

  • Liu L, Borkird C, Sung ZR (1985) Molecular cloning of cDNAs complementary to poly(A)+ mRNA differentially expressed during somatic embryogenesis. In: Terzi M, Pitto L, Sung ZR (eds) Proc somatic embryogenesis pp 70–76 IPRA. Roma, Italy

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Miyashita N, Langley CH (1988) Molecular and phenotypic variation of the white locus region in Drosophila melanogaster. Genetics 120:199–212

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479

    Google Scholar 

  • Pikaard CS, Brusca JS, Haunapel DJ, Park WD (1987) The two classes of genes for the major potato tuber protein, potatin, are differentially expressed in tubers and roots. Nucleic Acids Res 15:1979–1994

    Google Scholar 

  • Proudfoot NJ, Brownlee GG (1976) 3′ Non-coding region sequences in eukaryotic messenger RNA. Nature 263:211–214

    Google Scholar 

  • Pustell J, Kafatos FC (1984) A convenient and adaptable package of computer programs for DNA and protein sequence management, analysis and homology determination. Nucleic Acids Res 12:643–655

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sharon N, Lis H (1979) Comparative biochemistry of plant glycoproteins. Biochem Soc Trans 7:783–799

    Google Scholar 

  • Smith J, Furtner I, Sung ZR (1981) Nutritional and karyotypic characterization of a haploid cell culture of Daucus carota. In Vitro 17:315–321

    Google Scholar 

  • Smith JA, Krauss MR, Borkird C, Sung ZR (1988) A nuclear protein associated with cell divisions in plants. Planta 174:462–472

    Google Scholar 

  • Steward FC, Mapes MO, Smith J (1958) Growth and organized development of cultured cells. I. Growth and division of freely suspended cells. Am J Bot 45:693–703

    Google Scholar 

  • Sung ZR (1976) Turbidimetric determination of plant cell culture growth. Plant Physiol 57:460–462

    Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets; procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • von Heijne G (1985) Signal sequences The limits of variation. J Mol Biol 184:99–105

    Google Scholar 

  • von Heijne G (1986) A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 14:4683–4690

    Google Scholar 

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

    Google Scholar 

  • Watson MEE (1984) Compilation of published signal sequences. Nucleic Acids Res 12:5145–5164

    Google Scholar 

  • Wienbauer K, Herrero JJ, Filipowicz W (1988) Nuclear pre-mRNA in plants: distinct modes of 3′-splice-site selection in plants and animals. Mol Cell Biol 8:2042–2051

    Google Scholar 

  • Young RA, Davis RW (1983) Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci USA 80:1194–1198

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by J. Schell

Rights and permissions

Reprints and permissions

About this article

Cite this article

Franz, G., Hatzopoulos, P., Jones, T.J. et al. Molecular and genetic analysis of an embryonic gene, DC 8, from Daucus carota L.. Mol Gen Genet 218, 143–151 (1989). https://doi.org/10.1007/BF00330577

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330577

Key words

Navigation