Stable transformation of sunflower (Helianthus annuus L.) using a non-meristematic regeneration protocol and green fluorescent protein as a vital marker.
Stable transformation of sunflower was achieved using a non-meristematic hypocotyl explant regeneration protocol of public inbred HA300B. Uniformly transformed shoots were obtained after co-cultivation with Agrobacterium tumefaciens carrying a gfp (green fluorescent protein) gene containing an intron that blocks expression of gfp in Agrobacterium. Easily detectable, bright green fluorescence of transformed tissues was used to establish an optimal regeneration and transformation procedure. By Southern blot analysis, integration of the gfp and nptll genes was confirmed. Stable transformation efficiency was 0.1%. From 68 T1 plants analyzed, 17 showed transmission of transgene DNA and 15 of them contained the intact gfp gene. Expression of gfp was detected in 10 T1 plants carrying the intact gfp gene using a fluorimetric assay or western blot analysis. Expression of the nptll gene was confirmed in 13 T1 plants. The transformation system enables the rapid transfer of agronomically important genes.
Müller
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Institute of Plant Physiology and Biotechnology (260), University of Hohenheim, Stuttgart, Germany.
Iser
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M
Hess
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D015415:Biomarkers; D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000379:methods; D018506:Gene Expression Regulation, Plant; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006368:Helianthus / Q000235:genetics*; D019868:Kanamycin Kinase / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018519:Meristem; D018520:Plant Shoots / Q000166:cytology / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D012038:Regeneration; D012641:Selection, Genetic; D014170:Transformation, Genetic
D015415:Biomarkers; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D019868:Kanamycin Kinase