R. Mohr's research while affiliated with University of Freiburg and other places

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Publications (24)


Fig. 1. Phage and plaque assay characteristics. Transmission electron micrographs of (A) A-HIS1 and (B) A-HIS2. Scale bar = 50 nm. The same plaque assay of phage A-HIS1 after (C) 4 and (D) 9 days. (E) Confluent lysis of a A. marina MBIC11017 lawn by phage A-HIS1, which yielded spontaneous colony growth 1–2 months after infection. 
Fig. 2. Comparative genome map of A-HIS1 versus A-HIS2. Predicted ORFs are coloured according to the homology group of the Acaryochloris phage genes: purple, phage; pink, bacterial; blue, eukaryotic. Hypothetical ORFs are coloured in black where no annotation has been assigned based on database analyses. Grey lines represent matches using TBLASTX and the corresponding key denotes the associated e-values. A scale is included in kilobases. All ORFs shown encode predicted proteins. VSP indicates a virion structural protein, which was identified by mass spectrometry. (VSP) denotes an ORF categorized as a putative A-HIS1 structural protein based on amino acid sequence similarity to a corresponding ORF in A-HIS2 identified as a putative VSP by mass spectrometry and vice versa. (MS) denotes an ORF, which was identified by both BLASTP and mass spectrometry. See Table 1A and B for ORF details. 
Fig. 3. Identification of phage structural proteins. Coomassie-stained SDS-PAGE of purified proteins from A-HIS1 and A-HIS2 phage ghosts. Arrows indicate the position of bands removed from the gel and the corresponding ORFs, which were recognized after amino acid sequence identification. The putative major capsid and tape measure protein have been labelled in the figure. Other ORFs are annotated as virion structural proteins. 
Genomic and proteomic characterization of two novel siphovirus infecting the sedentary facultative epibiont cyanobacterium A caryochloris marina: Characterization of Acaryochloris phages
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December 2014

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355 Reads

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15 Citations

Environmental Microbiology

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Peter J Wheatley

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Acaryochloris marina is a symbiotic species of cyanobacteria that is capable of utilising far-red light. We report the characterisation of the phages A-HIS1 and A-HIS2, capable of infecting Acaryochloris. Morphological characterisation of these phages places them in the family Siphoviridae. However, molecular characterisation reveals that they do not show genetic similarity with any known siphoviruses. Whilst the phages do show synteny between each other, the nucleotide identity between the phages is low at 45-67 %, suggesting they diverged from each other some time ago. The greatest number of genes shared with another phage (a myovirus infecting marine Synechococcus) was four. Unlike most other cyanophages and in common with the Siphoviridae infecting Synechococcus no photosynthesis-related genes were found in the genome. CRISPR spacers from the host Acaryochloris had partial matches to sequences found within the phages, which is the first time CRISPRs have been reported in a cyanobacterial/cyanophage system. The phages also encode a homologue of the proteobacterial RNase T. The potential function of RNase T in the markup or digestion of crRNA hints at a novel mechanism for evading the host CRISPR system. This article is protected by copyright. All rights reserved.

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Draft Genome Sequence of the Filamentous Cyanobacterium Leptolyngbya sp. Strain Heron Island J, Exhibiting Chromatic Acclimation

February 2014

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68 Reads

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15 Citations

Genome Announcements

Leptolyngbya sp. strain Heron Island is a cyanobacterium exhibiting chromatic acclimation. However, this strain has strong interactions with other bacteria, making it impossible to obtain axenic cultures for sequencing. A protocol involving an analysis of tetranucleotide frequencies, G+C content, and BLAST searches has been described for separating the cyanobacterial scaffolds from those of its cooccurring bacteria.


Draft Genome Sequence of the Filamentous Cyanobacterium Leptolyngbya sp. Strain Heron Island J, Exhibiting Chromatic Acclimation

January 2014

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16 Reads

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6 Citations

Leptolyngbya sp. strain Heron Island is a cyanobacterium exhibiting chromatic acclimation. However, this strain has strong interactions with other bacteria, making it impossible to obtain axenic cultures for sequencing. A protocol involving an analysis of tetranucleotide frequencies, G+C content, and BLAST searches has been described for separating the cyanobacterial scaffolds from those of its cooccurring bacteria.


FIG. 1. Phylogenetic relationships amongst family A DNA polymerases. Unrooted Bayesian phylogenetic tree constructed in MrBayes. Nodes are labeled by a circle (support 5 1.00) or with support values. Where direct labeling was not possible, the corresponding support values are placed next to the two labels. Scale bar: amino acid substitutions per site. *denotes previously aligned sequences (Filée et al. 2002). Nodes labeled AxORFy refer to sequences from CAMERA (see supplementary table S3, Supplementary Material online). Numbers in parentheses refer to sequence details provided in supplementary table S4, Supplementary Material online.  
FIG. 2. Expression of Acaryochloris phage genes. Absolute gene copy number to estimate transcript abundance (converted from C T values) of Acaryochloris phage genes (A) mtDNA polymerase c: A-HIS1 ORF 14 and A-HIS2 ORF 20; (B) putative major capsid: A-HIS1 ORF 73 and A-HIS2 ORF 82; and (C) graph for A-HIS1 as in (B) replotted for clarity. Phages were infected at a MOI 5 1 at time zero. For A-HIS1, n 5 2 and A-HIS2, n 5 3. Error bars are one standard deviation.  
Discovery of Cyanophage Genomes Which Contain Mitochondrial DNA Polymerase

February 2011

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563 Reads

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18 Citations

Molecular Biology and Evolution

DNA polymerase γ is a family A DNA polymerase responsible for the replication of mitochondrial DNA in eukaryotes. The origins of DNA polymerase γ have remained elusive because it is not present in any known bacterium, though it has been hypothesized that mitochondria may have inherited the enzyme by phage-mediated nonorthologous displacement. Here, we present an analysis of two full-length homologues of this gene, which were found in the genomes of two bacteriophages, which infect the chlorophyll-d containing cyanobacterium Acaryochloris marina. Phylogenetic analyses of these phage DNA polymerase γ proteins show that they branch deeply within the DNA polymerase γ clade and therefore share a common origin with their eukaryotic homologues. We also found homologues of these phage polymerases in the environmental Community Cyberinfrastructure for Advanced Microbial Ecology Research and Analysis (CAMERA) database, which fell in the same clade. An analysis of the CAMERA assemblies containing the environmental homologues together with the filter fraction metadata indicated some of these assemblies may be of bacterial origin. We also show that the phage-encoded DNA polymerase γ is highly transcribed as the phage genomes are replicated. These findings provide data that may assist in reconstructing the evolution of mitochondria.


Figure 5: Cultures of Acaryochloris sp. HICR111A appear yellow-green (left flask), and those of A. marina MBIC11017 blue-green (right flask). (b) Light microscopy, (c) fluorescence microscopy and (d) field emission gun scanning electron microscopy (FEGSEM) of aggregated Acaryochloris sp. HICR111A. (e) Light microscopy, (f) fluorescence microscopy and (g) FEGSEM of non-aggregated A. marina MBIC11017. Both cultures were grown under medium white light at 25 °C.
Figure 5 Pigment ratios from A. marina cells (left side) compared with Acaryochloris sp. HICR111A cells (right side) grown under white light of low (5-10 mmol photons m À2 s À1 ), medium (25-30 mmol photons m À2 s À1 ) and high (75-100 mmol photons m À2 s À1 ) intensity. Left y axis describes Chl a/d (dark grey bars) and Pheo a/Chl d (light grey bars) ratios, and right y axis describes the Chl d/Zea (striped bars) ratio. The data shown are the average of three (A. marina) and two (Acaryochloris sp. HICR111A) experiments with standard errors. Pigment amounts were measured after separation by HPLC.
Figure 7 Transmission electron microscopy images of A. marina (a) and Acaryochloris sp. HICR111A (b).
Figure 8 Phylogenetic tree (neighbour joining (NJ)) based on 16S rRNA sequence comparison (1363 bp alignment length; all alignments available upon request) showing the relationship of Acaryochloris HICR111A with other members of the species complex and the position of Acaryochloris within the cyanobacterial kingdom. The comparison contains 39 cyanobacterial taxa, and the green sulphur bacterium Chlorobaculum tepidum as out group. Minimal evolution (ME) and NJ support values with Jukes-Cantor distances, and 10 000 bootstrap replicates were calculated using molecular evolutionary genetics analysis (MEGA) software version 4.0 (MEGA 4; Tamura et al., 2007). Bayesian posterior probability (PP) values were calculated using MrBayes 3.1.2. (Huelsenbeck and Ronquist, 2001; Ronquist and Huelsenbeck, 2003). NJ, ME and PP support values are shown in the table next to the tree indicating strong support for most nodes.
Figure 12: Phylogenetic tree based on 376 bp of the 16S rRNA gene (uc cb, uncultured cyanobacterium; uc Ac, uncultured Acaryochloris). Capital letters indicate the sample origin; A, red algae, Japan; B, Pseudoaxinella flava (sponge)-associated, Bahamas; C, Lyngbya-associated (Fern Hammick Spring, Florida); and D, Atacama Desert, Bolivia. The evolutionary history was inferred by neighbour joining (NJ; Saitou and Nei, 1987). The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) is shown next to the branches.
A new chlorophyll d-containing cyanobacterium: Evidence for niche adaptation in the genus Acaryochloris

November 2010

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394 Reads

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66 Citations

The ISME Journal

Chlorophyll d is a photosynthetic pigment that, based on chemical analyses, has only recently been recognized to be widespread in oceanic and lacustrine environments. However, the diversity of organisms harbouring this pigment is not known. Until now, the unicellular cyanobacterium Acaryochloris marina is the only characterized organism that uses chlorophyll d as a major photopigment. In this study we describe a new cyanobacterium possessing a high amount of chlorophyll d, which was isolated from waters around Heron Island, Great Barrier Reef (23° 26' 31.2″ S, 151° 54' 50.4″ E). The 16S ribosomal RNA is 2% divergent from the two previously described isolates of A. marina, which were isolated from waters around the Palau islands (Pacific Ocean) and the Salton Sea lake (California), suggesting that it belongs to a different clade within the genus Acaryochloris. An overview sequence analysis of its genome based on Illumina technology yielded 871 contigs with an accumulated length of 8 371 965 nt. Their analysis revealed typical features associated with Acaryochloris, such as an extended gene family for chlorophyll-binding proteins. However, compared with A. marina MBIC11017, distinct genetic, morphological and physiological differences were observed. Light saturation is reached at lower light intensities, Chl d/a ratios are less variable with light intensity and the phycobiliprotein phycocyanin is lacking, suggesting that cyanobacteria of the genus Acaryochloris occur in distinct ecotypes. These data characterize Acaryochloris as a niche-adapted cyanobacterium and show that more rigorous attempts are worthwhile to isolate, cultivate and analyse chlorophyll d-containing cyanobacteria for understanding the ecophysiology of these organisms.







Citations (5)


... Heron Island, Leptolyngbya boyana PCC 6306, and Leptolyngbya sp. KIOST-1 (19,27,36,37). The chromosomal GC content of Tl. ...

Reference:

Nitrogen Fixation Genes and Nitrogenase Activity of the Non-Heterocystous Cyanobacterium Thermoleptolyngbya sp. O-77
Draft Genome Sequence of the Filamentous Cyanobacterium Leptolyngbya sp. Strain Heron Island J, Exhibiting Chromatic Acclimation
  • Citing Article
  • January 2014

... The first reported Cyanostylovirus P-SS2 has a Pope et al. (2007) relatively large genome (108 kb) and can integrate it into the genome of Prochlorococcus MIT9313 (Sullivan et al., 2009). Though belonging to a lysogenic type, A-HIS1 targeting Acaryochloris marina MBIC11017 possesses a short latent period of 5 h (Chan et al., 2015). S-CBS1 infecting Synechococcus sp. ...

Genomic and proteomic characterization of two novel siphovirus infecting the sedentary facultative epibiont cyanobacterium A caryochloris marina: Characterization of Acaryochloris phages

Environmental Microbiology

... The PolA family has been explored in the past using various phylogenetic analyses. Nevertheless, such classifications are either outdated (25), or focus only on a particular subfamily (26,27) or a particular group of biological entities (28)(29)(30). These methods rely heavily on multiple sequence alignments (MSAs), which are challenged by the divergent nature of very large datasets of sequences and vary in ac-curacy depending on the algorithm used, demanding specialized solutions for different cases (31,32). ...

Discovery of Cyanophage Genomes Which Contain Mitochondrial DNA Polymerase

Molecular Biology and Evolution

... Microbialite is a benthic sedimentary deposit made of carbonate mud (particle diameter < 5 μm) that is formed with the mediation of microbes. Chlorophyll f has been shown to support some of the roles in photosynthetic reactions, in both the energy transfer and in the charge separation processes (Dunning, 2018;Follana-Berná, 2021 Chlorophyll d is used by a cyanobacterium known as Acaryochloris marina and is a phototroph that uses chlorophyll d as its principal light-harvesting pigment instead of chlorophyll a, the form commonly found in plants, algae and other cyanobacteria; and makes use of far-red (near-infrared) light for photosynthesis (Kühl et al, 2005;Chen, 2011;Mohr et al. 2010), see figure 7. ...

A new chlorophyll d-containing cyanobacterium: Evidence for niche adaptation in the genus Acaryochloris

The ISME Journal