Submitted to: Plant Journal
Publication Type: Peer reviewed journal
Publication Acceptance Date: 6/27/2007
Publication Date: 11/1/2007
Publication URL: www3.interscience.wiley.com/cgi-bin/fulltext/118488735/PDFSTART
Citation: Boavida, L.C., McCormick, S.M. 2007. Temperature as a determinant factor for increased and reproducible in vitro pollen germination in Arabidopsis thaliana. Plant Journal. 52(3):570-582. Interpretive Summary: Pollen germination is frequently used as an assay for reproductive function. In vitro pollen germination is easy for some species, such as tomato and tobacco, but very erratic and poor with other species, such as the model plant Arabidopsis. Here we describe a robust protocol for in vitro germination of Arabidopsis. The critical factors are incubation medium and temperature.
Technical Abstract: Despite much effort, a robust protocol for in vitro germination of Arabidopsis thaliana pollen was still elusive. Here we show that controlled temperatures, a largely disregarded factor in previous studies, and a simple optimized medium, solidified or liquid, yielded pollen germination rates above 80% and pollen tube lengths of hundreds of microns, with both Columbia and Landsberg erecta (L er ) ecotypes. We found that pollen germination and tube growth was dependent on pollen density in both liquid and solid medium. Pollen germination rates were not substantially affected by flower or plant age. The quartet1 mutation negatively affected pollen germination, especially in the Ler ecotype. This protocol will facilitate functional analyses of insertional mutants affecting male gametophyte function, and should allow detailed gene expression analyses during pollen tube growth. Arabidopsis thaliana can now be included on the list of plant species that are suitable models for physiological studies of pollen tube elongation and tip growth.