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Plant Cell Reports

, Volume 15, Issue 5, pp 311–316 | Cite as

Transgenic plantlets of ‘Chancellor’ grapevine (Vitis sp.) from biolistic transformation of embryogenic cell suspensions

  • Julie R. Kikkert
  • Dominique Hébert-Soulé
  • Patricia G. Wallace
  • Michael J. Striem
  • Bruce I. Reisch
Article

Abstract

Transgenic plantlets of ‘Chancellor’ grapevine (Vitis L. complex interspecific hybrid) were produced via biolistic transformation. Embryogenic cell suspensions were bombarded with 1 μm tungsten particles coated with pBI426 which encodes a fusion peptide between β-glucuronidase (GUS) and neomycin phosphotransferase II (NPTII). The fusion peptide is under the control of a double 35S Cauliflower Mosaic Virus promoter and a leader sequence from Alfalfa Mosaic Virus. The cells were placed on kanamycin-containing media (10, 25 or 50 mg/l) 2 d after bombardment. Activated charcoal reduced cell browning. Embryos were first observed on selective media 14–29 weeks after bombardment. More than 1600 clusters of embryos were germinated and/or assayed for GUS. Of 621 embryos assayed for GUS expression, 182 (29.3%) were positive. PCR confirmed the presence of the NPTII gene in all 5 GUS-positive and 2 GUS-negative (bombarded) embryos tested. In germination experiments, 15% of the embryo clusters produced at least one plant with normal shoot growth. Of 164 normal plants assayed for GUS expression, 37 (22.6%) were positive. The NPTII gene was amplified by PCR in 1 (of 1) GUS-positive and 4 (of 5) GUS-negative bombarded plants, but not in non-bombarded control plants. Southern blotting confirmed integration of the NPTII gene in all 3 of the GUS and PCR-NPTII positive plants tested. Biolistics is an efficient method for transformation of ‘Chancellor’ and should be applicable to other important grape cultivars.

Keywords

Activate Charcoal NPTII Gene Fusion Peptide Tungsten Particle Grape Cultivar 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

AC

activated charcoal

GUS

β-glucuronidase

2,4-D

2,4-dichlorophenoxyacetic acid

BA

6-benzylaminopurine

NAA

α-naphthalene acetic acid

TDZ

thidiazuron

NPTII

neomycin phosphotransferase II

Km

kanamycin

MS

Murashige and Skoog (1962) medium

WPM

Woody Plant Medium of Lloyd and McCown (1980)

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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Julie R. Kikkert
    • 1
  • Dominique Hébert-Soulé
    • 2
  • Patricia G. Wallace
    • 1
  • Michael J. Striem
    • 1
  • Bruce I. Reisch
    • 1
  1. 1.Department of Horticultural SciencesNew York State Agricultural Experiment Station, Cornell UniversityGenevaUSA
  2. 2.Institut National de la Recherche Agronomique, Station de Génétique et d'Amélioration des Plantes de MontpellierMauguioFrance

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