Abstract
Somatic embryogenesis can be synchronized by enriching carrot (Daucus carota L.) suspension cultures for small, dense clusters of cells termed proembryogenic masses (PEMs). Gene-expression programs of PEMs were compared with those of embryonic and mature tissues by in-vitro translation of representative mRNA populations and by nucleic-acid hybridization. Analysis of invitro-translated polypeptides by two-dimensional polyacrylamide gel electrophoresis revealed striking similarities between the mRNA populations of PEM and torpedo-stage embryos; substantial differences, however, were observed when in-vitro translation products of PEMs and torpedo embryos were compared with those of hypocotyls and leaves. Northern blots of RNA isolated from PEMs, staged embryos, and mature carrot tissues were hybridized with cDNA probes for Dc3, Dc5 and Dc13; these cDNA recombinants represent mRNAs that are regulated during carrot somatic embryogenesis. The pattern of expression of these embryo-regulated transcripts was similar in PEMs and somatic embryos but differed in other carrot tissues. These results indicate that many of the molecular processes of embryogenesis are already established in PEMs in the presence of auxin. Additional experiments indicate the utility of Dc3 as a molecular marker for the acquisition of embryogenic potential.
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Abbreviations
- cDNA:
-
complementary DNA
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- nt(s):
-
nucleotide(s)
- PAGE:
-
polyacrylamide gel electrophoresis
- PEM:
-
proembryogenic mass
- poly(A)RNA:
-
polyadenylated RNA
References
Allen, R.D., Nessler, C.L., Thomas, T.L. (1985) Developmental expression of sunflower 11S storage protein genes. Plant Mol. Biol. 5, 165–173
Aviv, H., Leder, P. (1977) Purification of biologically active globin mRNA by chromatography on oligothymidilic acidcellulose. Proc. Natl. Acad. Sci. USA 69, 1408–1412
Borkird, C., Choi, J.H., Sung, Z.R. (1986) Effect of 2,4-dichloro phenoxyacetic acid on th expression of embryogenic program in carrot. Plant Physiol. 81, 1143–1146
Choi, J.H., Liu, L-S., Borkird, C., Sung, Z.R. (1987) Isolation of cDNA clones for rare embryo-specific antigens in carrot cell cultures. Proc. Natl. Acad. Sci. USA 84, 1906–1910
Finer, J.J. (1983) The physiology of somatic embryogenesis in carrot (Daucus carota L.) and cotton (Gossypium klotz schianum Anderss.) PhD Dissertation, Texas A & M University, College Station
Fujimura, T., Komamine, A. (1979) Synchronization of somatic embryogenesis in a carrot cell suspension culture. Plant Physiol. 64, 162–164
Giuliano, G.D., Rosellini, D., Terzi, M. (1983) A new method for the purification of the different stages of carrot embryoids. Plant Cell Res. 2, 216–218
Glisin, V., Crkvenjakov, R., Byus, C. (1974) Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13, 2633–2640
Halperin, W. (1966) Alternative morphogenetic events in cell suspensions. Am. J. Bot. 53, 443–453
Halperin, W., Jensen, W.A. (1967) Ultrastructural changes during growth and embryogenesis in carrot cell cultures. J. Ultrastruct. Res. 18, 428–443
Huynh, T.V., Young, R.A., Davis, R.W. (1985) Constructing and screening cDNA libraries in λgt10 and λgt11 In: DNA cloning techniques: A practical approach, pp. 49–78, Glover, D.M., ed. IRL Press, London
LoSchiavo, F. (1984) A critical review of the procedures for embryo purification. Plant Mol. Biol. Reporter 2, 15–182
LoSchiavo, F., Giovinazzo, G., Terzi, M. (1983) 8-azaguanine resistant carrot cell mutants and their use as universal hybridizers. Mol. Gen. Genet. 192, 326–329
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Nomura, K., Komamine, A. (1985) Identification and isolation of single cells that produce somatic embryos at a high frequency in carrot suspension cultures. Plant Physiol. 79, 988–991
Nomura, K., Komamine, A. (1986) Molecular mechanisms of somatic embryogenesis. Oxford Surv. Plant Mol. Cell Biol. 3, 456–466
O'Farrell, P.H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021
Shah, D.M., Hightower, R.C., Meagher, R.B. (1982) Complete nucleotide sequence of a soybean actin gene. Proc. Natl. Acad. Sci. USA 79, 1022–1066
Thomas, P.S. (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. USA 77, 5201–5206
Thomas, T., Wilde, D. (1985) Analysis of gene expression in carrot somatic embryos. In: Somatic embryogenesis, pp. 77–85, Terzi, M., Pitto, L., Sung, Z.R., eds. IPRA, Rome
Thomas, T.L., Wilde, D. (1987) Analysis of carrot somatic embryo gene expression programs. Proc. VI Int. Congress. Plant Cell and Tissue Culture, pp. 83–93, Alan R. Liss, New York
Vergara, M.R., Biasini, G., LoSchiavo, F., Terzi, M. (1982) Isolation and characterization of carrot cell mutants to α-amanitin. Z. Pflanzenphysiol. 107, 313–319
Warren, G.S., Fowler, M. (1977) Physical method for the separation of various stages in the embryogenesis of carrot cell cultures. Plant Sci. Lett. 9, 71–76
Wilde, D. (1988) Patterns of gene expression during carrot somatic embryogenesis. Ph. D. Dissertation, Texas A & M University, College Station
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Wilde, H.D., Nelson, W.S., Booij, H. et al. Gene-expression programs in embryogenic and non-embryogenic carrot cultures. Planta 176, 205–211 (1988). https://doi.org/10.1007/BF00392446
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DOI: https://doi.org/10.1007/BF00392446