title: Transient expression and analysis of fluorescent reporter proteins in plant pollen tubes.
abstract: The pollen tube is an excellent single-cell model system for studying cellular processes in plant cell biology. This protocol describes a detailed step-by-step procedure with optimized conditions for introducing various fluorescent reporter proteins into lily, tobacco and Arabidopsis pollen grains by means of biolistics for their transient expression and subsequent analysis in germinating pollen tubes. The whole experiment consists of four major stages: coating gold microcarriers with DNA constructs, preparation of pollen grains, transformation of plasmid DNA into pollen grains by particle delivery system and germination of bombarded pollen grains in optimized germination media to obtain pollen tubes for protein trafficking, protein localization, drug treatment and organelle dynamics analysis. This protocol takes about 4-12 h from pollen preparation to protein detection.
authors.affiliation: School of Life Sciences, Centre for Cell and Developmental Biology, The Chinese University of Hong Kong, Hong Kong, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.forename: Liwen
authors.initials: L
authors.lastname: Jiang
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D018525:Germination; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D027762:Lilium / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010940:Plant Proteins / Q000032:analysis*; D053205:Pollen Tube / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins