Agrobacterium tumefaciens-mediated transformation of embryogenic tissue and transgenic plant regeneration in Chamaecyparis obtusa Sieb. et Zucc

A genetic transformation procedure for Chamaecyparis obtusa was developed after co-cultivation of embryogenic tissues with disarmed Agrobacterium tumefaciens strain C58/pMP90, which harbours the sgfp (synthetic green fluorescent protein) visual reporter and nptII (neomycin phoshotransferase II) sele...

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Bibliographic Details
Published in:Plant cell reports 2005-03, Vol.23 (12), p.796-802
Main Authors: Taniguchi, T, Kurita, M, Ohmiya, Y, Kondo, T
Format: Article
Language:English
Subjects:
Agrobacterium tumefaciens
Agrobacterium tumefaciens - genetics
Bacteria
Biomarkers
Cells, Cultured
Chamaecyparis - embryology
Chamaecyparis - genetics
Chamaecyparis obtusa
Coculture Techniques
conifers
Gene Expression Regulation, Plant - genetics
Gene Transfer Techniques
Genes, Reporter
genetic transformation
genetic vectors
Genetic Vectors - genetics
Genome, Plant
Green Fluorescent Proteins - genetics
Kanamycin Kinase - genetics
Microbiology
micropropagation
molecular sequence data
nucleotide sequences
plant regeneration
Plants, Genetically Modified - embryology
Plants, Genetically Modified - genetics
Regeneration - genetics
Seeds - embryology
Seeds - genetics
somatic embryogenesis
somatic embryos
Transformation, Genetic - genetics
Transgenes - genetics
Transgenic plants
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Summary:A genetic transformation procedure for Chamaecyparis obtusa was developed after co-cultivation of embryogenic tissues with disarmed Agrobacterium tumefaciens strain C58/pMP90, which harbours the sgfp (synthetic green fluorescent protein) visual reporter and nptII (neomycin phoshotransferase II) selectable marker genes. The highest transformation frequency was 22.5 independent transformed lines per dish (250 mg embryogenic tissue) following selection on kanamycin medium. Transgenic plantlets were regenerated through the maturation and germination of somatic embryos. The intensity of GFP fluorescence, observed under a fluorescence microscope, varied from very faint to relatively strong, depending on the transgenic line or part of the transgenic plant. The integration of the genes into the genome of regenerated plantlets was confirmed by Southern blot analysis.
ISSN:0721-7714
1432-203X
DOI:10.1007/s00299-004-0895-7