Remus-Emsermann et al.
Supplementary information
Supplemental Figure 1: Efficiency of cuticle lifts in the removal of phylloplane bacteria.
mCherry tagged Sphingomonas Fr1 were inoculated onto gnotobiotically grown
Arabidopsis and were allowed to grow for several weeks before cuticle tape lifts using
Yamori 50 were performed on the abaxial side of the leaves. A) Leaves before cuticle lifts
were performed B) Non-random images of bacteria that remained on leaves after cuticle
lifts were performed C) Bacteria recovered from leaves using cuticle tape lifts.
Remus-Emsermann et al.
Supplementary information
Supplemental Figure 2: False color images of PFA-fixed bacteria hybridized with FISH
probes. Three images of the same field of view were acquired in phase contrast and two
FISH probe channels (the general bacterial probe EUB338 and one group-specific probe).
The right panels show an overlay of all channels. A) alphaproteobacterium Sphingomonas
FR1,
probe
ALF698a;
B)
betaproteobacterium
Variovorax,
probe
Bet42a;
C)
gammaproteobacterium Pseudomonas syringae pv. syringae DC3000, probe Gam42a; D)
bacteroidetes Pedobacterium A03, probes CFB319a and CFB719a; E) actinobacterium
Arthrobacter, probe HGC689a. Scale bar = 10 µm F) Synthetic mix of pure culture bacteria
Remus-Emsermann et al.
Supplementary information
hybridized to group-specific FISH probes: Red: Arthrobacter sp. hybridized with HGC689aCy3; Blue: Pedobacter sp. A03 hybridized with Bet42a-Cy5; Green: Sphingomonas sp. Fr1
hybridized with ALF968-FAM; Cyan: Pseudomonas syringae DC3000 hybridized with
Gam42a-Cy5 and Gam42a-FAM; Pink: Variovorax sp. hybridized with CFB319a-Cy5,
CFP719-Cy5, CFB319a-Cy3 and CFB719-Cy3
Supplemental Figure 3: False color images of PFA-fixed bacteria hybridized to FISH
probes. Methylobacteria were grown under day/night regime prior to the FISH
hybridization. Four images of the same field of view were acquired in phase contrast; two
FISH probe channels (general bacterial probe EUB338 and the class-specific probe
ALF698a), and the infrared fluorescence channel indicative for AAnP. The right panels
show an overlay of all channels. A) Methylobacterium L04; B) Methylobacterium P01; C)
Methylobacterium PA1; D) Methylobacterium Q12. Scale bar = 10 µm