Advertisement

Improved Genetic Transformation of Cork Oak (Quercus suber L.)

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 877)

Abstract

An Agrobacterium-mediated transformation system for selected mature Quercus suber L. trees has been established. Leaf-derived somatic embryos in an early stage of development are inoculated with an AGL1 strain harboring a kanamycin-selectable plasmid carrying the gene of interest. The transformed embryos are induced to germinate and the plantlets transferred to soil.

This protocol, from adult cork oak to transformed plantlet, can be completed in about one and a half years. Transformation efficiencies (i.e., percentage of inoculated explants that yield independent transgenic embryogenic lines) vary depending on the cork oak genotype, reaching up to 43%.

Key words

AGL1 Agrobacterium tumefaciens Cork oak Fagaceae Herbicide resistance Kanamycin resistance Quercus suber Somatic embryogenesis Tree genetic transformation 
This is a preview of subscription content, log in to check access.

References

  1. 1.
    FAO (2004) Preliminary review of biotechnology in forestry, including genetic modification. Forest Genetic Resources Working Paper FGR/59E. Forest Resources Development Service, Forest Resources Division. Rome, ItalyGoogle Scholar
  2. 2.
    Boerjan W (2005) Biotechnology and the domestication of forest trees. Curr Opin Biotechnol 16:159–166PubMedCrossRefGoogle Scholar
  3. 3.
    Merkle SA et al (2007) Restoration of threatened species: a noble cause for transgenic trees. Tree Genet Genomes 3:111–118CrossRefGoogle Scholar
  4. 4.
    Álvarez R et al (2004) Genetic transformation of selected mature cork oak (Quercus suber L.) trees. Plant Cell Rep 23:218–223PubMedCrossRefGoogle Scholar
  5. 5.
    Sánchez N et al (2005) Agrobacterium-mediated transformation of cork oak (Quercus suber L.) somatic embryos. New For 29:169–176CrossRefGoogle Scholar
  6. 6.
    Álvarez R, Ordás RJ (2007) Improved genetic transformation protocol for cork oak (Quercus suber L.). Plant Cell Tiss Organ Cult 91:45–52CrossRefGoogle Scholar
  7. 7.
    Carraway DT et al (1994) Somatic embryogenesis and gene transfer in American chestnut. J Am Chestnut Found 8:29–33Google Scholar
  8. 8.
    Fernando DD et al (2006) In vitro germination and transient GFP expression of American chestnut (Castanea dentata) pollen. Plant Cell Rep 25:450–456PubMedCrossRefGoogle Scholar
  9. 9.
    Polin LD et al (2006) Agrobacterium-mediated transformation of American chestnut (Castanea dentata (Marsh.) Borkh.) somatic embryos. Plant Cell Tiss Organ Cult 84:69–79CrossRefGoogle Scholar
  10. 10.
    Rothrock RE et al (2007) Plate flooding as an alternative Agrobacterium-mediated transformation method for American chestnut somatic embryos. Plant Cell Tiss Organ Cult 88:93–99CrossRefGoogle Scholar
  11. 11.
    Andrade GM et al (2009) Sexually mature transgenic American chestnut trees via embryogenic suspension-based transformation. Plant Cell Rep 28:1385–1397PubMedCrossRefGoogle Scholar
  12. 12.
    Seabra R, Pais MS (1998) Genetic transformation of European chestnut. Plant Cell Rep 17:177–182CrossRefGoogle Scholar
  13. 13.
    Seabra R, Pais MS (1999) Genetic transformation of European chestnut (Castanea sativa Mill.) with genes of interest. Acta Hort (ISHS) 494:407–414Google Scholar
  14. 14.
    Corredoira E et al (2004) Agrobacterium-mediated transformation of European chestnut embryogenic cultures. Plant Cell Rep 23:311–318PubMedCrossRefGoogle Scholar
  15. 15.
    Corredoira E et al (2006) Genetic transformation of Castanea sativa Mill. by Agrobacterium tumefaciens. Acta Hort (ISHS) 693:387–394Google Scholar
  16. 16.
    Corredoira E et al (2007) Improving genetic transformation of European chestnut and cryopreservation of transgenic lines. Plant Cell Tiss Organ Cult 91:281–288CrossRefGoogle Scholar
  17. 17.
    Caro LA et al (2003) Agrobacterium rhizogenes vs auxinic induction for in vitro rhizogenesis of Prosopis chilensis and Nothofagus alpina. Biocell 27:311–318PubMedGoogle Scholar
  18. 18.
    Roest S et al (1991) Agrobacterium-mediated transformation of oak (Quercus robur L.). Acta Hort (ISHS) 289:259–260Google Scholar
  19. 19.
    Wilhelm E et al (1996) Plantlet regeneration via somatic embryogenesis and investigations on Agrobacterium tumefaciens mediated transformation of oak (Quercus robur). In: Ahuja MR, Boerjan W, Neale DB (eds) Somatic cell genetics and molecular genetics of trees. Kluwer Academic Publishers, DordrechtGoogle Scholar
  20. 20.
    Vidal N et al (2010) Regeneration of transgenic plants by Agrobacterium-mediated transformation of somatic embryos of juvenile and mature Quercus robur. Plant Cell Rep 29:1411–1422PubMedCrossRefGoogle Scholar
  21. 21.
    Aronson J et al (2009) Cork oak woodlands on the edge: ecology, adaptive management, and restoration. Island, Washington, DCGoogle Scholar
  22. 22.
    Paleo UF (2010) The dehesa/montado landscape. In: Bélair C, Ichikawa K, Wong BYL, Mulongoy KJ (eds). Secretariat of the Convention on Biological Diversity, Montreal. Technical Series No 52Google Scholar
  23. 23.
    Neumann KH (2006) Some studies on somatic embryogenesis: a tool in plant biotechnology. In: Sopory SK, Roy S, Kumar A (eds) Plant biotechnology. IK International Publishing House Pvt Ltd, New DelhiGoogle Scholar
  24. 24.
    Rose RJ et al (2010) Developmental biology of somatic embryogenesis. In: Pua EC, Davey MR (eds) Plant developmental biology - Biotechnological perspectives. Springer, BerlinGoogle Scholar
  25. 25.
    Bueno MA et al (1992) Plant regeneration through somatic embryogenesis in Quercus suber. Physiol Plant 85:30–34CrossRefGoogle Scholar
  26. 26.
    Manzanera J et al (1993) Somatic embryo induction and germination in Quercus suber L. Silvae Genet 42:90–93Google Scholar
  27. 27.
    Fernández-Guijarro B et al (1994) Somatic embryogenesis in Quercus suber L. In: Pardos JA, Ahuja MR, Elena-Rossello R (eds) Investigación Agraria, Sistemas y Recursos Forestales. INIA, MadridGoogle Scholar
  28. 28.
    Fernández-Guijarro B et al (1995) Influence of external factors on secondary embryogenesis and germination in somatic embryos from leaves of Quercus suber L. Plant Cell Tiss Organ Cult 41:99–106CrossRefGoogle Scholar
  29. 29.
    Hernández I et al (2001) Cloning mature cork oak (Quercus suber L.) trees by somatic embryogenesis. Melhoramento 37:50–57Google Scholar
  30. 30.
    Hernández I et al (2003) Vegetative propagation of Quercus suber L. by somatic embryogenesis. I. Factors affecting the induction in leaves from mature cork oak trees. Plant Cell Rep 21:759–764PubMedGoogle Scholar
  31. 31.
    Hernández I et al (2003) Vegetative propagation of Quercus suber L. by somatic embryogenesis. II. Plant regeneration from selected cork oak trees. Plant Cell Rep 21:765–770PubMedGoogle Scholar
  32. 32.
    Hernández I et al (2009) Growth data from a field trial of Quercus suber plants regenerated from selected trees and from their half-sib progenies by somatic embryogenesis. Acta Hort (ISHS) 812:493–498Google Scholar
  33. 33.
    Loureiro J et al (2005) Assessment of ploidy stability of the somatic embryogenesis process in Quercus suber L. using flow cytometry. Planta 221:815–822PubMedCrossRefGoogle Scholar
  34. 34.
    Lopes T et al (2006) Determination of genetic stability in long-term somatic embryogenic cultures and derived plantlets of cork oak using microsatellite markers. Tree Physiol 26:1145–1152PubMedCrossRefGoogle Scholar
  35. 35.
    Valladares S et al (2006) Plant regeneration through somatic embryogenesis from tissues of mature oak trees: true-to-type conformity of plantlets by RAPD analysis. Plant Cell Rep 25:879–886PubMedCrossRefGoogle Scholar
  36. 36.
    Fernandes P et al (2008) Cryopreservation of Quercus suber somatic embryos by encapsulation-dehydration and evaluation of genetic stability. Tree Physiol 28:1841–1850PubMedCrossRefGoogle Scholar
  37. 37.
    Pintos B et al (2007) Antimitotic agents increase the production of doubled-haploid embryos from cork oak anther culture. J Plant Physiol 164:1595–1604PubMedCrossRefGoogle Scholar
  38. 38.
    Pintos B et al (2008) Synthetic seed production from encapsulated somatic embryos of cork oak (Quercus suber L.) and automated growth monitoring. Plant Cell Tiss Organ Cult 95:217–225CrossRefGoogle Scholar
  39. 39.
    Álvarez R et al (2007) Cork oak trees (Quercus suber L.). In: Wang K (ed) Agrobacterium protocols, Volume II. Humana, TotowaGoogle Scholar
  40. 40.
    Álvarez R et al (2009) Genetic transformation of cork oak (Quercus suber L.) for herbicide resistance. Biotechnol Lett 31:1477–1483PubMedCrossRefGoogle Scholar
  41. 41.
    Pintos B et al (2010) Oak somatic and gametic embryos maturation is affected by charcoal and specific aminoacids mixture. Ann For Sci 67:205CrossRefGoogle Scholar
  42. 42.
    Lazo GR et al (1991) A DNA transformation-competent Arabidopsis genomic library in Agrobacterium. Bio/Technology 9:963–967PubMedCrossRefGoogle Scholar
  43. 43.
    Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  44. 44.
    Gamborg OL (1966) Aromatic metabolism in plants. II. Enzymes of the shikimate pathway in suspension cultures of plant cells. Biochem Cell Biol 44:791–799CrossRefGoogle Scholar
  45. 45.
    Schenk RU, Hildebrandt AC (1972) Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 50:199–204CrossRefGoogle Scholar
  46. 46.
    An G et al (1988) Binary vectors. In: Gelvin SB, Schilperoort RA, Verma DPS (eds) Plant molecular biology manual. Kluwer Academic Publishers, DordrechtGoogle Scholar
  47. 47.
    Sommer HE et al (1975) Differentiation of plantlets in longleaf pine (Pinus palustris Mill.) tissue cultured in vitro. Bot Gaz 136:196–200CrossRefGoogle Scholar
  48. 48.
    Toribio M et al (2005) Cork oak, Quercus suber L. In: Jain SM, Gupta PK (eds) Protocol for somatic embryogenesis in woody plants. Springer, DordrechtGoogle Scholar
  49. 49.
    Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  50. 50.
    Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene system. Plant Mol Biol Rep 5:387–405CrossRefGoogle Scholar
  51. 51.
    Humara J et al (1999) Improved efficiency of uidA gene transfer in stone pine (Pinus pinea) cotyledons using a modified binary vector. Can J For Res 29:1627–1632Google Scholar
  52. 52.
    Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res 12:8711–8721PubMedCrossRefGoogle Scholar
  53. 53.
    Berendzen K et al (2005) A rapid and versatile combined DNA/RNA extraction protocol and its application to the analysis of a novel DNA marker set polymorphic between Arabidopsis thaliana ecotypes Col-0 and Landsberg erecta. Plant Methods 1:4PubMedCrossRefGoogle Scholar
  54. 54.
    Gelvin SB (2003) Agrobacterium-mediated plant transformation: the biology behind the “gene-jockeying” tool. Microbiol Mol Biol R 67:16–37CrossRefGoogle Scholar
  55. 55.
    Álvarez R et al (2007) Agrobacterium protocols volume 2 Cork oak trees (Quercus suber L.). Springer, New York, NYGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Plant SciencesUniversity of CambridgeCambridgeUK
  2. 2.Departamento de Biología de Organismos y SistemasUniversidad de OviedoOviedoSpain

Personalised recommendations

Cite protocol

Buy options