Changes in secondary metabolic profiles of Microcystis aeruginosa strains in response to intraspecific interactions
Corresponding Author
Enora Briand
UMR CNRS 6553 ECOBIO, University of Rennes 1, 35042 Rennes Cedex, France
Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, 92093 USA
For correspondence. E-mail [email protected]; Tel. (+33) (0)2 23 23 57 13; Fax (+33) (0)2 23 23 50 26.Search for more papers by this authorMyriam Bormans
UMR CNRS 6553 ECOBIO, University of Rennes 1, 35042 Rennes Cedex, France
Search for more papers by this authorMuriel Gugger
Collection of Cyanobacteria, Institut Pasteur, 75724 Paris, France
Search for more papers by this authorPieter C. Dorrestein
Collaborative Mass Spectrometry Innovation Center, University of California San Diego, La Jolla, CA, 92093 USA
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, 92093 USA
Search for more papers by this authorWilliam H. Gerwick
Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, 92093 USA
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, 92093 USA
Search for more papers by this authorCorresponding Author
Enora Briand
UMR CNRS 6553 ECOBIO, University of Rennes 1, 35042 Rennes Cedex, France
Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, 92093 USA
For correspondence. E-mail [email protected]; Tel. (+33) (0)2 23 23 57 13; Fax (+33) (0)2 23 23 50 26.Search for more papers by this authorMyriam Bormans
UMR CNRS 6553 ECOBIO, University of Rennes 1, 35042 Rennes Cedex, France
Search for more papers by this authorMuriel Gugger
Collection of Cyanobacteria, Institut Pasteur, 75724 Paris, France
Search for more papers by this authorPieter C. Dorrestein
Collaborative Mass Spectrometry Innovation Center, University of California San Diego, La Jolla, CA, 92093 USA
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, 92093 USA
Search for more papers by this authorWilliam H. Gerwick
Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, 92093 USA
Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA, 92093 USA
Search for more papers by this authorSummary
The cyanobacteria Microcystis proliferate in freshwater ecosystems and produce bioactive compounds including the harmful toxins microcystins (MC). These secondary metabolites play an important role in shaping community composition through biotic interactions although their role and mode of regulation are poorly understood. As natural cyanobacterial populations include producing and non-producing strains, we tested if the production of a range of peptides by coexisting cells could be regulated through intraspecific interactions. With an innovative co-culturing chamber together with advanced mass spectrometry (MS) techniques, we monitored the growth and compared the metabolic profiles of a MC-producing as well as two non-MC-producing Microcystis strains under mono- and co-culture conditions. In monocultures, these strains grew comparably; however, the non-MC-producing mutant produced higher concentrations of cyanopeptolins, aerucyclamides and aeruginosins than the wild type. Physiological responses to co-culturing were reflected in a quantitative change in the production of the major peptides. Using a MS/MS-based molecular networking approach, we identified new analogues of known classes of peptides as well as new compounds. This work provides new insights into the factors that regulate the production of MC and other secondary metabolites in cyanobacteria, and suggests interchangeable or complementary functions allowing bloom-forming cyanobacteria to efficiently colonize and dominate in fluctuating aquatic environments.
Supporting Information
Filename | Description |
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emi12904-sup-0001-fig_s1.eps181.5 KB | Fig. S1. Identification of Cl2-aeruginosin 684. The WT and MT strains produced an aeruginosin with a mass of m/z 685 Da. This compound presented an isotopic pattern corresponding to a dichlorinated peptide (detection of pseudo-molecular peaks of M + , M + 2 and M + 4 Da, top). Its fragmentation pattern was characteristic of the aeruginosins with two intense fragments peaks at m/z 140 and 122 Da indicative of the Choi-immonium ion and its dehydrated derivative (bottom, Welker et al., 2006). |
emi12904-sup-0002-fig_s2.eps147.6 KB | Fig. S2. Identification of three new structural related compounds (P 612, 646 and 680). Partial mass spectrum of NT strain showing three molecular ion peaks (m/z 613, 647 and 681) with a 34 Da increment indicative of non-, mono- and dichlorinated variants of a peptide. This was also supported by the detection of pseudo-molecular ion peaks (M + 2 Da) with increasing relative intensity, corresponding well to theoretical isotopic patterns for chlorinated peptides. |
emi12904-sup-0003-fig_s3.eps574.6 KB | Fig. S3. Identification of cyanopeptolins variants produced by WT and MT strains. MS/MS data of each variants: (A) Cya A, (B) Cya B, (C) Cya C, (D) Cya 895 and (E) Cya 963A. Grey bars indicate fragment series used to distinguish cyanopeptolins. |
emi12904-sup-0004-fig_s4.eps640.7 KB | Fig. S4. Identification of cyanopeptolins variants produced by NT strain. MS/MS data of each variant: (A) Cya 914, (B) Cya 948, (C) Cya 962, (D) Cya 964 and (E) Cya 978. Grey bars indicate fragment series used to distinguished cyanopeptolins. |
emi12904-sup-0005-fig_s5.eps238.1 KB | Fig. S5. Relative concentrations given as a fold change over WT in monoculture of selected intracellular compounds for WT, MT and MT with MC-LR under mono- (WT/WT, MT/MT) and co-culture (WT/MT) conditions and statistical results. MC: Microcystin, Arg: Aeruginosin, Cya: Cyanopeptolin, Aer: Aerucyclamide. |
emi12904-sup-0006-fig_s6.eps183.6 KB | Fig. S6. Correlation between the relative concentrations of major compounds produced in the WT/MT experiment determined by the addition of internal standard and by the spectral counts obtained for each compound and represented by the pie-chart. (A) Intracellular compounds and (B) extracellular compounds. |
emi12904-sup-0007-fig_s7.eps111 KB | Fig. S7. Over time diffusion of MC-LR through a 0.45 μm membrane in co-culture chambers. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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