Article

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. PolinAffiliated withFaculty of Forest and Natural Resources Management, State University of New York, College of Environmental Science and Forestry
  • , Haiying LiangAffiliated withSchool of Forest Resources, Department of Horticulture and Huck Institute for Life Sciences, Pennsylvania State University
  • , Ronald E. RothrockAffiliated withFaculty of Forest and Natural Resources Management, State University of New York, College of Environmental Science and Forestry
  • , Mutsumi NishiiAffiliated withFaculty of Environmental and Forest Biology, State University of New York, College of Environmental Science and Forestry
  • , Deborah L. DiehlAffiliated withFaculty of Forest and Natural Resources Management, State University of New York, College of Environmental Science and Forestry
  • , Andrew E. NewhouseAffiliated withFaculty of Environmental and Forest Biology, State University of New York, College of Environmental Science and Forestry
  • , C. Joseph NairnAffiliated withDaniel B. Warnell School of Forest Resources, University of Georgia
  • , William A. PowellAffiliated withFaculty of Environmental and Forest Biology, State University of New York, College of Environmental Science and Forestry
  • , Charles A. MaynardAffiliated withFaculty of Forest and Natural Resources Management, State University of New York, College of Environmental Science and Forestry Email author 

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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