The Production of Transgenic Scots Pine (Pinus Sylvestris L.) via the Application of Transformed Pollen in Controlled Crossings |
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Authors: | Aronen Tuija S. Nikkanen Teijo O. Häggman Hely M. |
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Affiliation: | (1) Finnish Forest Research Institute, Punkaharju Research Station, Finlandiantie 18, FIN-58450 Punkaharju, Finland;(2) Department of Biology, University of Oulu, PO Box 3000, FIN-90014 Oulu, Finland |
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Abstract: | The study demonstrates the production of a transgenic Scots pine (Pinus sylvestris L.) seedling through the application of transformed pollen in controlled crossings. The pollen lots were transformed by particle bombardment, resulting in transient transformation frequencies varying from 15 to 49% of the germinated pollen grains, and bombarded pollen was used to pollinate megasporangiate strobili. Progeny was screened by histochemical, GUS assays, and selected seedlings were further analysed by PCR. PCR amplification revealed the presence of both the nptII and gusA genes in one seedling (23/237). Results were confirmed by Southern blot analysis. The morphology and growth of this transgenic seedling was normal. Although the transformation frequency of recovered plants was very low (1/14999), the present protocol suggests that production of transgenic Scots pine is possible without the use of any tissue culture methods or the involvement of marker genes, for selection of transformants. |
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Keywords: | genetic transformation of pollen particle bombardment Pinus sylvestris transgenic pine |
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