Article Figures & Data
Figures
- FIG 1
Putative prokaryotic consortium IPPAS B-1204. 16S rRNA gene phylogeny of the sequenced strains and microscopic images are shown. (A) 16S rRNA gene phylogeny of Leptolyngbya sp. strain IPPAS B-1204. (B) 16S rRNA gene phylogeny of Porphyrobacter sp. strain IPPAS B-1204. Phylogenetic analysis based on 16S rRNA sequences extracted manually after PGAP annotation was conducted using the maximum likelihood method based on the Kimura 2-parameter model (15) in MEGA7 (16). Initial trees for the heuristic search were obtained automatically by applying the neighbor-join and BioNJ algorithms to a matrix of pairwise distances estimated using the maximum composite likelihood (MCL) approach and then selecting the topology with the superior log likelihood value. Bootstrap values of >60% are shown. Gloeobacter violaceus PCC 7421 and Rhodospirillum rubrum ATCC 11170 were used as outgroups for panels A and B, respectively. (C) Merged fluorescence image of the IPPAS B-1204 culture stained with DAPI (4′,6-diamidino-2-phenylindole) for DNA (blue), FM 1-43 for cellular membranes (green), and autofluorescence of chlorophyll (red). Bar = 5 μm. (D) Magnified filament of the Leptolyngbya sp. (the arrow indicates trichomes visualized by blue and red) closely interacting with the Porphyrobacter sp. (asterisk indicates individual cells outlined by membranes stained green). Bar = 5 μm. Images were acquired using three channels of an Axio Imager Z2 epifluorescence microscope equipped with an AxioCam MRm high-resolution monochrome charge-coupled-device (CCD) camera and merged using AxioVision v4.8 software (Carl Zeiss, Göttingen, Germany). For the first channel, filter set 49 was used (excitation G 365, emission BP 445/50), and epifluorescence images of DAPI-DNA complexes were assigned a blue pseudocolor; for the second channel, filter set 44 was used (excitation BP 475/40, emission BP 530/50), and images of cell membranes stained with FM 1-43 were assigned a green pseudocolor; for the third channel, filter set 45 was used (excitation BP 560/40, emission BP 630/75), and cyanobacterial chlorophyll autofluorescence was assigned a red pseudocolor. Scale bar = 5 μm.
