Abstract
A full-length cDNA for ECP31, an embryogenic cell protein from carrot (Daucus carota L.) with a M r of 31000 (Kiyosue T, Satoh S, Kamada H, Harada H (1991) Plant Physiol 95: 1077–1083), was isolated from a cDNA library prepared from embryogenic cells using PCR-amplified DNA as a probe. The genomic Southern blot analysis revealed that there are two or three genes for ECP31 in the carrot genome. The transcripts of ECP31 accumulated in the peripheral regions of clusters of embryogenic cells and disappeared in the course of somatic embryogenesis that was induced by transfer of the embryogenic cells to auxin-free media. The cDNA encodes a polypeptide of 256 amino acids, and the calculated molecular weight of this polypeptide is 26 111. The deduced amino acid sequence shows a high degree (62.2%) of similarity to that of a protein that is abundant during late embryogenesis of cotton (LEA D34; Baker JC, Steele C, Dure III (1988) Plant Mol Biol 11: 227–291). The mRNAs for ECP31 started to accumulate in zygotic embryos at a late stage of embryogenesis but were undetectable in mature embryos within 24 h after imbibition of seeds. In dry fruits (seeds), the transcripts were detected only in zygotic embryos by in situ hybridization. The level of ECP31 transcripts increased after treatment with abscisic acid (ABA) in torpedo-shaped somatic embryos but not in seven-day-old seedlings. These results suggest that both embryo-specific factor(s) and ABA are involved in the expression of the gene for ECP31.
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References
Aleith F, Richter G: Gene expression during induction of somatic embryogenesis in carrot cell suspensions. Planta 183: 17–24 (1990).
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K: In: Current Protocols in Molecular Biology, Chapter 4. Greene Publishing Associates/Wiley-Interscience, New York (1989).
Baker JC, Steele C, DureIII: Sequence and characterization of 6 Lea proteins and their genes from cotton. Plant Mol Biol 11: 227–291 (1988).
Borkird C, Sung ZR: Isolation and characterization of ABA-insensitive cell lines of carrot. Plant Physiol 84: 1001–1006 (1987).
Borkird C, Choi J, Jin Z, Franz G, Hatzopoulos P, Chorneau R, Bonas U, Pelegri F, Sung ZR: Developmental regulation of embryonic genes in plants. Proc Natl Acad Sci USA 85: 6399–6403 (1988).
Choi JH, Liu LS, Borkird C, Sung ZR: Cloning of genes developmentally regulated during plant embryogenesis. Proc Natl Acad Sci USA 84: 1906–1910 (1987).
Close TJ, Kortt AA, Chandler PM: A cDNA-based comparison of dehydration-induced proteins (dehydrins) in barley and corn. Plant Mol Biol 13: 95–108 (1989).
de Vries S, Booij H, Meyerink P, Huisman G, van Spanje M, Wilde D, Thomas TL, van Kammen A: Acquisition of embryogenic potential in carrot cell suspension cultures. Planta 176: 194–204 (1988).
DureIII L, Crouch M, Harada J, Ho T-H D, Mundy J, Quatrano R, Thomas T, Sung ZR: Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol Biol 12: 475–486 (1989).
Galau GA, Hughes DW, DureIII L: Abscisic acid induction of cloned cotton late embryogenesis-abundant (Lea) mRNAs. Plant Mol Biol 7: 155–170 (1986).
Goldberg RB, Barker SJ, Perez-Grau L: Regulation of gene expression during plant embryogenesis. Cell 56: 149–160 (1989).
Halperin W: Alternative morphogenetic events in cell suspensions. Am J Bot 53: 443–453 (1966).
Kiyosue T, Satoh S, Kamada H, Harada H: Purification and immunohistochemical detection of an embryogenic cell protein in carrot. Plant Physiol 95: 1077–1083 (1991).
Kiyosue T, Nakayama J, Satoh S, Isogai A, Suzuki Y, Kamada H, Harada H: Partial amino acid sequence of ECP31, a carrot embryogenic cell protein, and enhancement of its accumulation by abscisic acid in somatic embryos. Planta 186: 337–342 (1992).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning; A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Marcotte WR, Russell SH, Quatrano RS: Abscisic acid-responsive sequences from the Em gene of wheat. Plant Cell 1: 969–976 (1989).
Mundy J, Chua NH: Abscisic acid and water-stress induce the expression of a novel rice gene. EMBO J 7: 2279–2286 (1988).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
Nomura K, Komamine A: Molecular mechanisms of somatic embryogenesis. Oxf Surv Plant Mol Cell Biol 3: 456–466 (1986).
Satoh S, Kamada H, Harada H, Fujii T: Auxin-controlled glycoprotein release into the medium of embryogenic carrot cells. Plant Physiol 81: 931–933 (1986).
Skriver K, Mundy J: Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512 (1990).
Steward FC, Mapes MO, Smith J: Growth and organized development of cultured cells. I. Growth and division of freely suspended cells. Am J Bot 45: 693–703 (1958).
Sung ZR, Feinberg A, Chorneau R, Borkird C, Turner I, Smith J, Terzi M, LoSchiavo F, Giuliano G, Pitto L, Nuti-Ronchi V: Developmental biology of embryogenesis from carrot cultures. Plant Mol Biol Rep 2: 3–14 (1984).
Thomas TL, Wilde HD: Analysis of gene expression in carrot somatic embryos. In: Terzi J, Pitto L, Sung ZR (eds) Somatic Embryogenesis, pp 77–85. IPRA, Rome (1985).
Thomas TL, Wilde HD: Analysis of carrot somatic embryo gene expression programs. Proceedings of the VI-International Congress on Plant Cell and Tissue Culture, pp. 83–93 Alan R. Liss, New York (1987).
Wilde HD, Nelson WS, Booij H, de Vries SC, Thomas TL: Gene-expression programs in embryogenic and non-embryogenic carrot cultures. Planta 176: 205–211 (1988).
Yamaguchi-Shinozaki K, Mino M, Mundy J, Chua NH: Analysis of an ABA-responsive rice gene promoter in transgenic tobacco. Plant Mol Biol 15: 905–912 (1990).
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Kiyosue, T., Yamaguchi-Shinozaki, K., Shinozaki, K. et al. Isolation and characterization of a cDNA that encodes ECP31, an embryogenic-cell protein from carrot. Plant Mol Biol 19, 239–249 (1992). https://doi.org/10.1007/BF00027345
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DOI: https://doi.org/10.1007/BF00027345