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The sensitivity of transgenic spruce (Picea glauca (Moench) Voss) cotyledonary somatic embryos and somatic seedlings to kanamycin selection

Par Conseil national de recherches du Canada

DOITrouver le DOI : https://doi.org/10.1023/A:1018473604111
AuteurRechercher : 1; Rechercher : 1; Rechercher : 1; Rechercher : 1; Rechercher : 1
Affiliation
  1. Conseil national de recherches du Canada. Institut de biotechnologie des plantes du CNRC
FormatTexte, Article
Sujetβ-GUS; embryogenesis reinduction; kanamycin; NPTII; Piceaglauca; selection; somatic embryos; somatic seedlings; transformation
Résumé
Transformed white spruce cultures containing immature Stage I somatic embryos, were developed after particle bombardment of somatic embryos with pBI 426, carrying an expression cassette with a gus::nptII fusion gene. These Stage I cultures did not show tolerance to kanamycin concentrations above 3 to 5 mg l−1, although assays for GUS and NPTII showed that functional enzymes were present in the transgenic tissue. Embryonic liquid suspension cultures were initiated from this transformed tissue. After treatment on agar-solidified maturation medium with 48 μm (±)-ABA high numbers of Stage III (cotyledonary) somatic embryos were produced. When subjected to an embryogenesis re-induction process with 2,4-D and BA, these Stage III embryos produced a new generation of Stage I embryogenic tissues which could tolerate 5--10 mg l−1 kanamycin. Stage III somatic embryos could alternatively be placed onto germination medium for the development of somatic seedlings. When germinated in the presence of 20 mg l−1 kanamycin, 77% of inoculants were resistant. The stability of integration of the gus::nptII fusion gene in the genome of white spruce Stage III somatic embryos and somatic seedlings was confirmed through Southern blot analysis
Date de publication
Maison d’éditionSpringer Link
Dans
  • Transgenic Research 6, nº 2 (mars 1997) : 123–131.
Langueanglais
Publications évaluées par des pairsOui
Identificateur172466
NRCC-38934
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Identificateur de l’enregistrement6e664e6f-fa66-4b25-9298-ed967f503509
Enregistrement créé2023-11-01
Enregistrement modifié2023-11-01
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