title: Expression of green fluorescent protein in pollen of oilseed rape (Brassica napus L.) and its utility for assessing pollen movement in the field.
abstract: Transgene movement via pollen is an important component of gene flow from transgenic plants. Here, we present proof-of-concept studies that demonstrate the monitoring of short distant movement of pollen expressing a genetically encoded fluorescent tag in oilseed rape (Brassica napus L. cv. Westar). Transgenic oilseed rape plants were produced using Agrobacterium-mediated transformation method with the pBINDC1 construct containing a green fluorescent protein (GFP) variant, mGFP5-ER, under the control of the pollen-specific LAT59 promoter from tomato. Transgenic pollen was differentiated from non-transgenic pollen in vivo by a unique spectral signature, and was shown to be an effective tool to monitor pollen movement in the greenhouse and field. GFP-tagged pollen also served as a practical marker to determine the zygosity of plants. In a greenhouse pollen flow study, more pollen was captured at closer distances from the source plant plot with consistent wind generated by a fan. Under field conditions, GFP transgenic pollen grains were detected up to a distance of 15 m, the farthest distance from source plants assayed. GFP-tagged pollen was easily distinguishable from non-transgenic pollen using an epifluorescence microscope.
authors.affiliation: University of Tennessee, Department of Plant Sciences, Knoxville, TN 37996, USA.
authors.forename: Hong S
authors.initials: HS
authors.lastname: Moon
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Halfhill
authors.forename: Laura C
authors.initials: LC
authors.lastname: Hudson
authors.forename: Reginald J
authors.initials: RJ
authors.lastname: Millwood
authors.forename: C Neal
authors.initials: CN
authors.lastname: Stewart
mesh_terms: D029688:Brassica napus / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D015336:Molecular Probe Techniques; D009038:Motion; D011058:Pollen / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D012639:Seeds / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins