Abstract
Stable genetic transformation of black spruce (Picea mariana) and tamarack (Larix laricina) was obtained via microprojectile bombardment of tissues from two stages of somatic embryogenesis (mature cotyledonary somatic embryos and suspension from embryonal masses) using the Biolistic PDS-1000/He device. A total of 14 transgenic lines were obtained with the two species with the vectors pRT99GUS and pBI426 using low level kanamycin selection. The integration of the foreign genes was confirmed by Southern hybridization and complex patterns were observed. Transgenic seedlings were regenerated for both species and expression of the GUS gene observed in needles. Two chimeric genes containing components from black spruce genes were used for transient gene expression into conifer tissues; a cDNA (MBF1) coding for a myb C1 related gene and a chalcone synthase promoter. Both components were proven functional when reintroduced into their original genome and in L. laricina tissues. Furthermore, the chimeric MBF1 gene was functional when introduced into corn cell suspension. These results indicate the evolutionary conservation of the genes involved in the anthocyanin pathways in gymnosperms and angiosperms. Additionally, the Green Fluorescent Protein gene from a jellyfish (Aequorea victoria) was evaluated as a marker for genetic transformation in conifer tissues.
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References
Bodeau, J.P., and V. Walbot (1992) Regulated transcription of the maize Bronze-2 promoter in electroporated protoplasts requires the C1 and R gene products. Mol Gen Genet 233:379–387.
Bommineni, V.R., Chibbar, R.N., Datla, R.S.S., and Tsang, E.W.T. (1993) Transformation of white spruce (Picea glauca) somatic embryos by microprojectile bombardment. Plant Cell Rep 13:17–23.
Chalfie, M., Yuan, T., Euskirchen, G., Ward, W.W., and Prasher, D.C. (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805.
Charest, P.J., Calero, N., Lachance, D., Mitsumune, M., and Yoo, B.Y. (1993a) The use of microprojectile DNA delivery to bypass the long life cycle of tree species in gene expression studies. Curr. Topics Bot. Res. 1:151–163.
Charest, P.J., Calero, N., Lachance, D., Datla, R.S.S., Duchesne, L.C., and Tsang, E.W.T. (1993b) Microprojectile DNA delivery in conifer species: factors affecting assessment of transient gene expression using the ß-glucuronidase reporter gene. Plant Cell Rep. 12:189–193.
Charest, P.J., Devantier, Y., and Lachance D. (1995) Stable genetic transformation of Picea mariana (black spruce) via microprojectile bombardement. In Vitro Cell Dev Biol (Plant) (Submitted).
Charest, P.J. and Michel, M.F. (1991) Basics of plant genetic engineering and potential applications to tree species. Petawawa National Forestry Institute, Information Report PI-X-104. 48pp.
Cheliak, W.M. and Klimaszewska, K.K. (1991) Genetic variation in somatic embryogenic response in open-pollinated families of black spruce. Theor. Appl. Genet. 82:185–190.
Datla, R.S.S., Hammerlindl, J.K., Pelcher, L.E., Crosby, W.L., and Selvaraj, G. (1991) A bifunctional fusion between ß-glucuronidase and neomycin phosphotransferase: a broad-spectrum marker enzyme for plants. Gene 101:239–246.
Datla, R.S.S., Bekkaoui, F., Hammerlindl, J.K., Pilate, G., Dunstan, D.I., and Crosby, W.L. (1993) Improved high-level constitutive foreign gene expression in plants using an AMV RNA4 untranslated leader sequence. Plant Sci. 94:139–149.
Ellis, D.D., McCabe, D.E., Mclnnis, S., Ramachandran, R., Rüssel, D.R., Wallace, K.M., Martinell, B.J., Roberts, D.R., Raffa, K.F., and McCown; B.H. (1993) Stable transformation of Picea glauca by particle acceleration. BioTechnol. 11: 84–89.
Huang, Y., Diner, A.M., and Kamosky, D.F. Agrobacteriutn rhizogenes-mediated genetic transformation and regeneration of a conifer: Larix decidua. In Vitro Cell Dev. Biol. 27:201–207.
Jefferson, R.A., Kavanagh, T.A., and Bevan, M.W. (1987) GUS fusion: ß-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6:3901–3907.
Jones, J.D.G., Shlumukov, L., Carland, F., English, J., Scofield, S.R., Bishop, G.J., and Harrison, K. (1992) Effective vectors for transformation, expression of heterologous genes, and assaying transposon excision in transgenic plants. Transgenic Res. 1:285–297.
Kay, R., Chan, A., Daly, M., and McPherson, J. (1987) Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236:1299–1302.
Klimazewska, K., Devantier, Y., Lachance, D., Lelu, M.A., and Charest, P.J. (1995) Larix laricina (tamarack):somatic embryogenesis and genetic transformation, (in preparation)
Lelu, M.A., Klimaszewska, K., Jones, C., Ward, C., von Aderkas, P., and Charest, P.J. (1993) A laboratory guide to somatic embryogenesis in spruce and larch. Petawawa National Forestry Institute. Information report PI-X-111. 57pp..
Robertson, D., Weissinger, A. K., Ackley, R., Glover, S., and Sederoff, R. (1991) Genetic transformation of Norway spruce (Picea abies (L.) Karst) using somatic embryo expiants by microprojectile bombardment. Plant Mol. Biol. 19:925–935.
Seguin, A., Lachance, D., and Charest, P.J. (1995) Transient gene expression and stable genetic transformation into conifer tissues by microprojectile bombardment. Plant Tiss. Cult. Man. (in press).
Topfer, R., Schell, J., and Steinbiss, H.-H. (1988) Versatile cloning vectors for transient gene expression and direct gene transfer in plant cells. Nucleic Acid Res. 16:8725.
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Charest, P.J. et al. (1996). Stable Genetic Transformation in Black Spruce and Tamarack and the Transgenic Expression of Conifer Genes. In: Ahuja, M.R., Boerjan, W., Neale, D.B. (eds) Somatic Cell Genetics and Molecular Genetics of Trees. Forestry Sciences, vol 49. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-3983-0_13
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DOI: https://doi.org/10.1007/978-94-011-3983-0_13
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