Transient gene expression in pine pollen tubes following particle bombardment |
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| Authors: | D D Fernando J N Owens S Misra |
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| Institution: | (1) Department of Environmental and Forest Biology, State University of New York, 1 Forestry Drive, Syracuse, NY 13210, USA, US;(2) Centre for Forest Biology, University of Victoria, Victoria, British Columbia V8W 3N5, Canada, CA;(3) Department of Microbiology and Biochemistry, University of Victoria, Victoria, British Columbia V8W 3P6, Canada, CA |
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| Abstract: | A biolistic particle delivery system was used to genetically transform pollen tubes of three species of white pine (Pinus aristata, P. griffithii and P. monticola). The introduced plasmid DNA contained the GUS coding sequence flanked by the 35S CaMV promoter and NOS terminator sequences.
Successful gene delivery was demonstrated by transient GUS expression as evaluated by standard histochemical assay. Distance
of target specimens significantly influenced transient GUS expression in all three species of white pine. A target distance
of 6 cm resulted in a significant number of transformed pollen tubes in P. aristata and P. griffithii, while distances of 6 and 9 cm resulted in a significant number of transformed pollen tubes in P. monticola. Generally, the number of pollen tubes expressing GUS activity was higher in P. aristata than in P. griffithii and P. monticola. The possibility of using GUS-transformed pollen tubes in conjunction with in vitro fertilization in conifers was examined.
Gene expression in pollen tubes was also examined under electron microscopy where the X-glu reaction product occurred as large
crystalline electron-dense precipitates in the cytoplasm.
Received: 17 December 1998 / Revision received: 17 March 1999 / Accepted: 14 April 1999 |
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| Keywords: | Pinus aristata P griffithii P monticola Pollen tubes Transformation Particle bombardment In vitro fertilization |
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