Wounding by bombardment yields highly efficient Agrobacterium-mediated transformation of carnation (Dianthus caryophyllus L.)

Amir Zuker, Asaph Ahroni, Tzvi Tzfira, Hagit Ben-Meir, Alexander Vainstein*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

41 Scopus citations

Abstract

Highly efficient Agrobacterium-mediated transformation of carnation (Dianthus caryophyllus L.) was obtained by first wounding stem explants via microprojectile bombardment. When this was followed by cocultivation with disarmed Agrobacterium in the dark, the transformation frequency-based on transient GUS expression-increased to over 10-fold that of explants wounded by other means and cocultivated under constant light. Two cycles of regeneration/selection on kanamycin were employed to generate stably transformed carnation plants and eliminate chimeras: first, plantlets were regenerated from inoculated stem explants and then leaves from these plantlets were used to generate transgenes in a second selection cycle of adventitious shoot regeneration. Agrobacterium strain AGLO, carrying the binary vector pCGN7001 containing uidA and nptII genes, was used in the stable transformation experiments. The combination of wounding via bombardment, cocultivation in the dark and two cycles of kanamycin selection yielded an overall transformation efficiency of 1-2 transgenes per 10 stem explants for the three carnation varieties analyzed. Histochemical and molecular analyses of marker genes in T0 and T1 generations confirmed the transgenic nature of the selected plants.

Original languageAmerican English
Pages (from-to)367-375
Number of pages9
JournalMolecular Breeding
Volume5
Issue number4
DOIs
StatePublished - 1999

Bibliographical note

Funding Information:
We wish to thank Shemi Ltd. (Shdema, Israel) and Mizpor Ltd. (Tquma, Israel) for providing the plant materials. This research was supported by Grant US-2233-92 from BARD, the United States-Israel Binational Agricultural Research and Development Fund, by the Ministry of Agriculture, and by the Association of Israeli Flower Growers.

Keywords

  • Genetic engineering
  • Kanamycin selection
  • Microprojectile bombardment
  • Stable transformation
  • Transgenic carnation

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