Plant Cell, Tissue and Organ Culture

, Volume 84, Issue 1, pp 69–79

Agrobacterium-mediated transformation of American chestnut (Castanea dentata (Marsh.) Borkh.) somatic embryos

  • Linda D. Polin
  • Haiying Liang
  • Ronald E. Rothrock
  • Mutsumi Nishii
  • Deborah L. Diehl
  • Andrew E. Newhouse
  • C. Joseph Nairn
  • William A. Powell
  • Charles A. Maynard
Article

Abstract

These studies were designed to test if a binary vector containing the gfp, bar and oxalate oxidase genes could transform American chestnut somatic embryos; to see if a desiccation treatment during co-cultivation would affect the transformation frequency of different American chestnut somatic embryo clones; to explore the effects of more rapid desiccation; and to see if the antibiotics used to kill the Agrobacterium were interfering with the regeneration of the somatic embryos. Two days of gradual desiccation was found to significantly enhance transient GFP expression frequency. When this treatment was tested on six American chestnut clones, five were transformed and four of these remained embryogenic. Transformation was confirmed by Southern hybridization. Phenotypically normal transgenic shoots were regenerated and rooted. Vascular tissue specific expression of the oxalate oxidase gene was detected in one transgenic line. Carbenicillin, cefotaxime, and tricarcillin were found to not interfere with the regeneration of transformed embryos.

Key words:

bar clonal variation desiccation gfp oxalate oxidase regeneration 

Abbreviations

2,4-D

2,4-dichlorophenoxyacetic acid

BA

6-benzyladenine

bar

bialaphos-resistance gene

Carb

carbenicillin

Cef

cefotaxime

mgfp5-ER

modified green fluorescent protein gene

OxO

oxalate oxidase

PCR

polymerase chain reaction

PPT

phosphinothricin

SDS

sodium dodecyl sulfate

SSC

sodium chloride sodium citrate

TE

Tris–EDTA

Tric

tricarcillin

WPM

McCown’s woody plant medium salts

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Carraway DT, Wilde HD, Merkle SA, (1994) Somatic embryogenesis and gene transfer in American chestnut J. Am. Chestnut Found. 8: 29–33Google Scholar
  2. Carraway DT, Merkle SA, (1997) Plantlet regeneration from somatic embryos of American chestnut Can. J. For. Res. 27: 1805–1812CrossRefGoogle Scholar
  3. Cheng M, Hu TC, Layton J, Liu CN, Fry JE, (2003) Desiccation of plant tissues post-Agrobacterium infection enhance T-DNA delivery and increases stable transformation efficiency in wheat In Vitro Cell. Dev. Biol. – Plant 39: 595–604CrossRefGoogle Scholar
  4. Connors BJ, Miller M, Maynard CA, Powell WA, (2002) American chestnut promoters capable of directing reporter gene expression in transgenic Arabidopsis plants Plant Sci. 163: 771–781CrossRefGoogle Scholar
  5. Corredoira E, Montenegro D, San-José MC, Viéitez AM, Ballester A, (2004) Agrobacterium-mediated transformation of European chestnut embryogenic cultures Plant Cell Rep. 23: 311–318PubMedGoogle Scholar
  6. Covert SF, Kapoor P, Lee M, Briley A, Nairn CJ, (2001) Agrobacterium tumefaciens-mediated transformation of Fusarium circinatum Mycol. Res. 105: 259–264CrossRefGoogle Scholar
  7. Desfeux C, Clough SJ, Bent AF, (2000) Female reproductive tissues are the primary target of Agrobacterium-mediated transformation by the Arabidopsis floral-dip method Plant Physiol. 123: 895–904CrossRefPubMedGoogle Scholar
  8. Diller JD, Clapper RB, (1965) A progress report on attempts to bring back the chestnut tree in the eastern United States, 1954–1964 J. For. 63: 186–188Google Scholar
  9. Dratewka-Kos E, Rahman S, Grzekzak ZF, Kennedy TD, Murray RK, Lane G, (1989) The polypeptide structure of germin as deduced from cDNA sequencing J. Biol. Chem. 264: 4896–4900PubMedGoogle Scholar
  10. Dumas B, Freyssinet G, Pallett KE, (1995) Tissue-specific expression of germin-like oxalate oxidase during development and fungal infection of barley seedlings Plant Physiol. 107: 1091–1096PubMedGoogle Scholar
  11. Garbino JE, Belknap WR, (1994) Isolation of a ubiquitin-ribosomal protein gene (ubi3) from potato and expression of its promoter in transgenic plants Plant Mol. Biol. 24: 119–127CrossRefPubMedGoogle Scholar
  12. Gonzales ML, Vieitez AM, Vieitez E, (1985) Somatic embryogenesis from chestnut cotyledon tissue cultured in vitro Sci. Hortic. 27: 97–103CrossRefGoogle Scholar
  13. Haseloff J, Siemering KR, (1998) The uses of GFP in plants. In: Chalfie M, Kain S, (eds) Green Fluorescent Protein: Properties, Applications, and Protocols Wiley-Liss, New York pp. 191–220Google Scholar
  14. Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT, (1985) A simple and general method for transferring genes into plants Science 227: 1229–1231ADSGoogle Scholar
  15. LaPierre SL (2003) Studies in American chestnut (Castanea dentata (Marsh.) Borkh.) micropropagation and acclimatization. M.S. thesis, State University of New York, College of Environmental Science and Forestry, Syracuse, NYGoogle Scholar
  16. Liang H, Maynard CA, Allen RD, Powell WA, (2001) Increased Septoria musiva resistance in transgenic hybrid poplar leaves expressing a wheat oxalate oxidase gene Plant Mol. Biol. 45: 619–629CrossRefPubMedGoogle Scholar
  17. Lloyd G, McCown B, (1980) Commercially feasible micropropagation of Mountain Laurel, Kalmia latifolia, by use of shoot tip culture Comb. Proc. Int. Plant Propagators Soc. 30: 421–427Google Scholar
  18. Lodhi MA, Ye GN, Weeden NF, Reisch BI, (1994) A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species Plant Mol. Biol. Rep. 12: 6–13CrossRefGoogle Scholar
  19. Mason HS, DeWald DB, Mullet JE, (1993) Identification of a methyl jasmonate-responsive domain in the soybean vspB promoter Plant Cell 5: 241–251CrossRefPubMedGoogle Scholar
  20. Maynard CA (1991) Using PCR to confirm Agrobacterium transformation of American chestnut (Castanea dentata). Abstract No. 927, In: Program and Abstracts of the Third International Congress of Plant Molecular Biology: Molecular Biology of Plant Growth and Development. Tucson, Arizona, October 6–11, 1991Google Scholar
  21. Merkle S, Carraway DT, Watson-Pauley BA & Wilde HD (1992) Somatic embryogenesis and gene transfer in American chestnut. In: Proceedings of the International Chestnut Conference. West Virginia University Press, Morgantown, West Virginia, July 10–14 1992Google Scholar
  22. Merkle SA, Wiecko AT, Watson-Pauley BA, (1991) Somatic embryogenesis in American chestnut Can. J. For. Res. 21: 1698–1701Google Scholar
  23. Mullins KV, Llewellyn DJ, Hartney VJ, Strauss S, Dennis ES, (1997) Regeneration and transformation of Eucalyptus camaldulensis Plant Cell Rep. 16: 787–791CrossRefGoogle Scholar
  24. Murrill WA, (1906) A serious chestnut disease J. New York Bot. Garden 7: 143–153Google Scholar
  25. Nitsch JP, Nitsch C (1969) Haploid plants from pollen grains. Science 163: 85–87Google Scholar
  26. Piagnani C, Eccher T, (1990) Somatic embryogenesis in chestnut Acta Hortic. 280: 159–161Google Scholar
  27. Polin LD (2004) Biolistic transformation of American chestnut (Castanea dentata (Marsh.) Borkh.) somatic embryos using green fluorescent protein as a scorable marker. M.S. thesis, State University of New York, College of Environmental Science and Forestry, Syracuse, NYGoogle Scholar
  28. Roane MK, Griffin GJ, Elkins JR, (1986) Chestnut blight, other Endothia diseases, and the genus Endothia. APS Press American Phytopathological Society, St. Paul, MNGoogle Scholar
  29. Robichaud RL, Lessard VC, Merkle SA, (2004) Treatments affecting maturation and germination of American chestnut somatic embryos Plant Physiol. 161: 957–969CrossRefPubMedGoogle Scholar
  30. Sambrook J, Russell DW, (2001) Molecular cloning: a laboratory manual 3 Cold Spring Harbor Laboratory Press Cold Spring Harbor, NYGoogle Scholar
  31. Sambrook J, Fritsch EE, Maniatis T, (1989) Molecular Cloning: A Laboratory Manual 2 Cold Spring Harbor Laboratory Press Cold Spring Harbor, NYGoogle Scholar
  32. Seabra RC, Paris MS, (1998) Genetic transformation of European chestnut Plant Cell Rep. 17: 177–182CrossRefGoogle Scholar
  33. Vieitez FJ, San-Jose MC, Ballester A, Vieitez AM, (1990) Somatic embryogenesis in cultured immature zygotic embryos in chestnut J. Plant Physiol. 136: 253–256Google Scholar
  34. Xing Z, Satchwell MF, Powell WA, Maynard CA, (1997) Micropropagation of American chestnut: increasing rooting rate and preventing shoot-tip necrosis In Vitro Cell. Dev. Biol. – Plant 33:43–48Google Scholar
  35. Xing Z, Powell WA, Maynard CA, (1999) Development and germination of somatic embryos in American chestnut Plant Cell Tiss. Org. Cult. 57: 47–55CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Linda D. Polin
    • 1
  • Haiying Liang
    • 4
  • Ronald E. Rothrock
    • 1
  • Mutsumi Nishii
    • 2
  • Deborah L. Diehl
    • 1
  • Andrew E. Newhouse
    • 2
  • C. Joseph Nairn
    • 3
  • William A. Powell
    • 2
  • Charles A. Maynard
    • 1
  1. 1.Faculty of Forest and Natural Resources ManagementState University of New York, College of Environmental Science and ForestrySyracuseUSA
  2. 2.Faculty of Environmental and Forest BiologyState University of New York, College of Environmental Science and ForestrySyracuseUSA
  3. 3.Daniel B. Warnell School of Forest ResourcesUniversity of GeorgiaAthensUSA
  4. 4.School of Forest Resources, Department of Horticulture and Huck Institute for Life SciencesPennsylvania State UniversityUniversity ParkUSA

Personalised recommendations