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

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1023/A:1018473604111
AuthorSearch for: 1; Search for: 1; Search for: 1; Search for: 1; Search for: 1
Affiliation
  1. National Research Council of Canada. NRC Plant Biotechnology Institute
FormatText, Article
Subjectβ-GUS; embryogenesis reinduction; kanamycin; NPTII; Piceaglauca; selection; somatic embryos; somatic seedlings; transformation
Abstract
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
Publication date
PublisherSpringer Link
In
  • Transgenic Research 6, no. 2 (March 1997): 123–131.
LanguageEnglish
Peer reviewedYes
Identifier172466
NRCC-38934
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Record created2023-11-01
Record modified2023-11-01
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