Pollen viability and transgene expression following storage in honey.
Transgenic plants of tobacco and Arabidopsis that produce genetically marked pollen, expressing the reporter gene uidA (gusA), were generated to determine whether pollen proteins can be expressed and stable in honey, a potential route by which foreign proteins might enter the wider environment. Hydrated tobacco pollen was found to lose viability rapidly in honey, while pollen in the natural dehydrated form remained viable for at least several days and in some cases several weeks, as determined by FDA staining activity and germinability. Dehydrated pollen was found to be capable of transient foreign gene expression, following microprojectile bombardment, after incubation in honey for at least 120 h. PCR amplification of transgene sequences in pollen of transgenic plants revealed that pollen DNA can remain relatively intact after 7 weeks in honey. GUS enzyme activity analysis and SDS-PAGE of pollen proteins revealed that foreign and native pollen proteins are stable in pollen incubated in honey for at least 6 weeks. We conclude that pollen may represent an ecologically important vector for transgenic protein products.
Eady
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C
Dept of Botany, University of Leicester, UK.
Twell
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D
Lindsey
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K
D017360:Arabidopsis / Q000235:genetics / Q000502:physiology; D001937:Brassica / Q000502:physiology; D018744:DNA, Plant / Q000378:metabolism; D005452:Fluoresceins / Q000737:chemistry; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D006722:Honey; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011058:Pollen / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014026:Nicotiana / Q000235:genetics / Q000502:physiology
D018744:DNA, Plant; D005452:Fluoresceins; D011993:Recombinant Fusion Proteins; D005966:Glucuronidase; C018506:diacetylfluorescein