Summary
Transgenic plants of three Picea species were produced after coculture of embryogenic tissue with the disarmed strain of Agrobacterium tumefaciens C58/pMP90/pBIV10 and selection on medium containing kanamycin. In addition to the nptII selectable gene (conferring resistance to kanamycin), the vector carried the uidA (β-glucuronidase) marker gene. Transformation frequencies were dependent on the species, genotype, and post-cocultivation procedure. Of the three species tested, P. mariana was transformed at the highest frequency, followed by P. glauca and P. abies. The transgenic state of the embryogenic tissue was initially, confirmed by histochemical β-glucuronidase (GUS) assay followed by Southern hybridization. One to over five copies of T-DNA were detected in various transgenic lines analyzed. Transgenic plants were regenerated for all species using modified protocols for maturation and germination of somatic embryos.
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References
Birch, R. G. Plant transformation: problems and strategies for practical application. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48:297–326 1997.
Campbell, R. A.; Durzan, D. J. Induction of multiple buds and needles in tissue cultures of Picea glauca. Can. J. Bot. 53:1652–1657; 1975.
Charest, P. J.; Devantier, Y.; Lachance, D. Stable genetic transformation of Picea mariana (black spruce) via particle bombardment. In Vitro Cell. Dev. Biol. Plant 32:91–99; 1996.
Clapham, D.; Demel, P.; Elfstrand, M.; Koop, H.-U.; Sabala, I.; von Arnold, S. Gene transfer by particle bombardment to embryogenic cultures of Picea abies and the production of transgenic plantlets. Scand. J. For. Res. 15:151–160; 2000.
Datla, R. S. S.; Bekkaoui, F.; Hammerlindl, J. K.; Pilate, G.; Dunstan, D. I.; Crosby, W. L. Improved high-level constitutive foreign gene expression in plants using an AMV RNA4 untranslated leader sequence. Plant Sci. 94:139–149; 1993.
Datla, R. S. S.; Hammerlindl, J. K.; Pelcher, L. E.; Crosby, W. L.; Selvaraj, G. A bifunctional fusion between β-glucuronidase and neomycin phosphotransferase: a broad-spectrum marker enzyme for plants. Gene 101:239–246; 1991.
De Buck, S.; De Wilde, C.; Van Montagu, M.; Depicker, A. Determination of the T-DNA transfer and T-DNA integration frequencies upon cocultivation of Arabidopsis thaliana root explants. Mol. Plant-Microbe Interact. 6:658–665; 2000.
De Neve, M.; De Buck, S.; Jacobs, A.; Van Montagu, M.; Depicker, A. T-DNA integration patterns in co-transformed plant cells suggest that T-DNA repeats originate from co-integration of separate T-DNAs. Plant J. 11:15–29; 1997.
Ellis, D. D. Transformation of gymnosperms.. In: Jain, S. M., Gupta, P. K., Newton, R. J., eds. Somatic embryogenesis in woody plants. Dordrecht, Kluwer Academic Publishers; 1995;227–247.
Ellis, D. D.; McCabe, D. E.; McInnis, S.; Ramachandran R.; Russell, D. R.; Wallace, K. M.; Martinell, B. J.; Roberts, D. R.; Raffa, K. F.; McCown, B. H. Stable transformation of Picea glauca by particle acceleration. Bio/Technology 11:84–89; 1993.
Garin, E.; Bernier-Cardou, M.; Isabel, N.; Klimaszewska, K.; Plourde, A. Effect of sugars, amino acids, and culture technique on maturation of somatic embryos of Pinus strobus on medium with two gellan gum concentrations. Plant Cell Tiss. Organ Cult. 62:27–37; 2000.
Holland, L.; Gemmell, J. E.; Charity, J. A.; Walter, C. Foreign gene transfer into Pinus radiata cotyledons by Agrobacterium tumefaciens N.Z.J. For. Sci. 27:289–304; 1997.
Huang, Y.; Diner, A. M.; Karnosky, D. F. Agrobacterium rhizogenes-mediated genetic transformation and regeneration of a conifer: Larix decidua In Vitro Cell. Dev. Biol. Plant 27:201–207; 1991.
Humara, J. M.; López, M.; Ordás, R. J. Agrobacterium tumefaciens-mediated transformation of Pinus pinea L. cotyledons: an assessment of factors influencing the efficiency of uidA gene transfer. Plant Cell Rep. 19:51–58; 1999a.
Humara, J. M.; Marin, M. S.; Parra, F.; Ordás, R. J. Improved efficiency of uidA gene transfer in stone pine (Pinus pinea) cotyledons using a modified binary vector. Can. J. For. Res. 29:1627–1632; 1999b.
Jefferson, R. A. Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5:387–405; 1987.
Klimaszewska, K.; Devantier, Y.; Lachance, D.; Lelu, M. A.; Charest, P. J. Larix laricina (tamarack): somatic embryogenesis and genetic transformation. Can. J. For. Res. 27:538–550; 1997.
Klimaszewska, K.; Park, Y.-S.; Overton, C.; MacEacheron, I.; Bonga, J. M. Optimized somatic embryogenesis in Pinus strobus L In Vitro Cell. Dev. Biol. Plant 37:392–399; 2001.
Klimaszewska, K.; Smith, D. R. Maturation of somatic embryos of Pinus strobus is promoted by a high concentration of gellan gum. Physiol. Plant 100:949–957; 1997.
Koncz, C.; Schell, J. The promoter of TL-DNA gene 5 controls the tissuespecific expression of chimaeric genes carried by a novel type of Argobacterium binary vector. Mol. Gen. Genet. 204:383–396.; 1986.
Lelu, M. A.; Pilate, C. Transgenic in Larix. In: Jain, S. M., Minocha, S. C., eds. Molecular biology of woody plants, vol 2. Dordrecht: Kluwe Academic Publisheres; 2000:119–134.
Levée, V.; Garin, E.; Klimaszewska, K.; Séguin, A. Stable genetic transformation of white pine (Pinus strobus L.) after cocultivation of embryogenic tissues with Agrobacterium tumefaciens. Mol. Breed. 5:429–440; 1999.
Levée, V.; Lelu, M.-A.; Jouanin, L.; Cornu, D.; Pilate, G. Agrobacterium tumefaciens-mediated transformation of hybrid blarch (Larix kaempferi XL decidua) and transgenic plant regeneration. Plant Cell Rep. 16:680–685; 1997.
Litvay, J. D.; Verma, D. C.; Johnson, M. A. Influence of a loblolly pine (Pinus taeda L.) culture medium and its components on growth and somatic embryogenesis of the wild carrot (Daucus carota L) Plant Cell Rep. 4:325–328; 1985.
Robertson, D.; Weissinger, A. K.; Ackley, R.; Glover, S.; Sederoff, R. R. Genetic transformation of Norway spruce (Picea abies (L.) Karst.) using somatic embryo explants by microprojectile bombardment. Plant Mol. Biol. 19:925–935; 1992.
Tepfer, M.; Casse-Delbart, F. Agrobacterium rhizogenes as the vector for transforming higher plants. Microbiol. Sci. 4:24–28; 1987.
Tian, L.-N.; Charest, P. J.; Séguin, A.; Rutledge, R. G. Hygromycin resistance is an effective selectable marker for biolistic transformation of black spruce (Picea mariana). Plant Cell Rep. 19:358–362; 2000.
Tzfira, T.; Yarnitzky, O.; Vainstein, A.; Altman, A. Agrobacterium rhizogenes-mediated DNA transfer in Pinus halepensis Mill. Plant Cell Rep. 16:26–31; 1996.
Walter, C.; Crace, L. J.; Donaldson, S. S.; Moody, J.; Gemmell, J. E.; van der Maas, S.; Kvaalen, H.; Lonneborg, A. An efficient Biolistic® transformation protocol for Picea abies embryogenic tissue and regeneration of transgenic plants. Can. J. For. Res. 29:1539–1546; 1999.
Walter, C.; Grace, L. J.; Wagner, A.; White, D. W. R.; Walden, A. R.; Donaldson, S. S.; Hinton, H.; Gardner, R. C.; Smith, D. R. Stable transformation and regeneration of transgenic plants of Pinus radiata D. Don. Plant Cell Rep. 17:460–468; 1998.
Wenck, A. R.; Quinn, M.; Whetten, R. W.; Pullman, G.; Sederoff, R. High-efficiency Agrobacterium-mediated transformation of Norway spruce (Picea abies) and loblolly pine (Pinus taeda). Plant Mol. Biol. 39:407–416; 1999.
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Klimaszewska, K., Lachance, D., Pelletier, G. et al. Regeneration of transgenic Picea glauca, P. Mariana, and P. abies after cocultivation of embryogenic tissue with Agrobacterium tumefaciens . In Vitro Cell.Dev.Biol.-Plant 37, 748–755 (2001). https://doi.org/10.1007/s11627-001-0124-9
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DOI: https://doi.org/10.1007/s11627-001-0124-9