title: 454 Sequencing of phyllosphere microbiota
design: Cells were washed from the leaf material by 3 min of alternate shaking, vortexing, and sonication. The cell suspen sion was separated from the leaf material by filtration through a nylon mesh (pore size 200 m; Spectrum Europe BV, Breda, Th e Netherlands). 6 mL of 80% Percoll (Sigma-Aldrich) was pipetted below the cell suspension, and the 50-mL tubes were centrif uged for 5 min at 800 g. The bacterial cell suspension above the Percoll layer was transferred into a fresh 50-mL tube and c ells were pelleted at 3,150 g for 15 min. Cell pellets from multiple tubes were pooled into 1.5 mL reaction tubes and washed twice  with TE-buffer plus Pefabloc SC. Cell pellets were immediately frozen at -20C.