title: An experimental method to facilitate the identification of hybrid sporophytes in the moss Physcomitrella patens using fluorescent tagged lines.
abstract: • The sequencing of the Physcomitrella patens genome, combined with the high frequency of gene targeting in this species, makes it ideal for reverse genetic studies. For forward genetic studies, experimental crosses and genetic analysis of progeny are essential. • Since P. patens is monoicous, producing both male and female gametes on the same gametophore, and undergoing self-fertilization at a high frequency, the identification of crossed sporophytes is difficult. Usually spores from many sporophytes from a mixed culture must be tested for the production of recombinant progeny. • Here, we describe the use of transgenic lines that express a fluorescent transgene constitutively, to provide a direct visual screen for hybrid sporophytes. • We show that segregations in crosses obtained with this technique are as expected, and demonstrate its utility for the study of the rate of outcrossing between three isolates of P. patens.
authors.affiliation: Department of Biology, Campus Box 1137, Washington University in St Louis, St Louis, MO 63130, USA.
authors.forename: Pierre-François
authors.initials: PF
authors.lastname: Perroud
authors.affiliation: Department of Biology, Campus Box 1137, Washington University in St Louis, St Louis, MO 63130, USA.|Centre for Plant Sciences, Leeds University, Leeds, LS2 9JT, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Cove
authors.affiliation: Department of Biology, Campus Box 1137, Washington University in St Louis, St Louis, MO 63130, USA.
authors.forename: Ralph S
authors.initials: RS
authors.lastname: Quatrano
authors.affiliation: Department of Biology, Box 118525, University of Florida, Gainesville, FL 32611, USA.
authors.forename: Stuart F
authors.initials: SF
authors.lastname: McDaniel
mesh_terms: D019068:Bryopsida / Q000235:genetics*; D003433:Crosses, Genetic; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006824:Hybridization, Genetic; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D030821:Plants, Genetically Modified / Q000378:metabolism; D019076:Transgenes
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins