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. 2022 Aug 31;10(4):e0181422.
doi: 10.1128/spectrum.01814-22. Epub 2022 Jun 30.

The Self-Bleaching Process of Microcystis aeruginosa is Delayed by a Symbiotic Bacterium Pseudomonas sp. MAE1-K and Promoted by Methionine Deficiency

Affiliations

The Self-Bleaching Process of Microcystis aeruginosa is Delayed by a Symbiotic Bacterium Pseudomonas sp. MAE1-K and Promoted by Methionine Deficiency

Jaejoon Jung et al. Microbiol Spectr. .

Abstract

Various interactions between marine cyanobacteria and heterotrophic bacteria have been known, but the symbiotic relationships between Microcystis and heterotrophic bacteria remain unclear. An axenic M. aeruginosa culture (NIES-298) was quickly bleached after exponential growth, whereas a xenic M. aeruginosa culture (KW) showed a normal growth curve, suggesting that some symbiotic bacteria may delay this bleaching. The bleaching process of M. aeruginosa was distinguished from the phenomena of previously proposed chlorosis and programmed cell death in various characteristics. Bleached cultures of NIES-298 quickly bleached actively growing M. aeruginosa cultures, suggesting that M. aeruginosa itself produces bleach-causing compounds. Pseudomonas sp. MAE1-K delaying the bleaching of NIES-298 cultures was isolated from the KW culture. Bleached cultures of NIES-298 treated with strain MAE1-K lost their bleaching ability, suggesting that strain MAE1-K rescues M. aeruginosa from bleaching via inactivation of bleaching compounds. From Tn5 transposon mutant screening, a metZ mutant of strain MAE1-K (F-D3) unable to synthesize methionine, promoting the bleaching of NIES-298 cultures but capable of inactivating bleaching compounds, was obtained. The bleaching process of NIES-298 cultures was promoted with the coculture of mutant F-D3 and delayed by methionine supplementation, suggesting that the bleaching process of M. aeruginosa is promoted by methionine deficiency. IMPORTANCE Cyanobacterial blooms in freshwaters represent serious global concerns for the ecosystem and human health. In this study, we found that one of the major species in cyanobacterial blooms, Microcystis aeruginosa, was quickly collapsed after exponential growth by producing self-bleaching compounds and that a symbiotic bacterium, Pseudomonas sp. MAE1-K delayed the bleaching process via the inactivation of bleaching compounds. In addition, we found that a metZ mutant of strain MAE1-K (F-D3) causing methionine deficiency promoted the bleaching process of M. aeruginosa, suggesting that methionine deficiency may induce the production of bleaching compounds. These results will provide insights into the symbiotic relationships between M. aeruginosa and heterotrophic bacteria that will contribute to developing novel strategies to control cyanobacterial blooms.

Keywords: Microcystis aeruginosa; Pseudomonas; bleaching; bleaching compounds; metZ mutant; methionine deficiency.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

FIG 1
FIG 1
Growth profiles of xenic (KW) and axenic (NIES-298) cultures of M. aeruginosa. Images showing colors of the xenic and axenic cultures during growth are displayed as insets inside the figure.
FIG 2
FIG 2
Microscopic analyses showing the general bleaching process of cells in the axenic NIES-298 culture. Differential interference contrast (A–D) and transmission electron (E–H) microscopic images. (A and E) General morphology of cells during the exponential growth phase. (B and F) General morphology of cells during the initial death phase. (C, D, G, and H) General morphology of cells during the late death phase.
FIG 3
FIG 3
Effects of NIES-298 cultures on the growth of strain NIES-298. The growth of NIES-298 cultures was evaluated in BG11 broth supplemented with different amounts of filtered NIES-298 cultures, which were prepared by filtering NIES-298 cultures at the exponential growth (A) and death (B) phases.
FIG 4
FIG 4
Effects of bacterial strains isolated from the xenic KW culture on the growth of axenic NIES-298 cultures.
FIG 5
FIG 5
Pseudomonas sp. MAE1-K with the ability to inactivate the bleaching property of bleached culture to NIES-298 culture. The bleached culture was prepared by filtering bleached NIES-298 culture at the end of the death phase. The growth of NIES-298 cultures was evaluated in BG11 broth supplemented with 5% (vol/vol) bleached culture or 5% MAE1-K-treated bleached culture.
FIG 6
FIG 6
A transposon mutant (F-D3) of strain MAE1-K losing the ability to delay the bleaching of NIES-298 culture. (A) Bleaching images of NIES-298 culture promoted by strain F-D3. Wild-type MAE1-K and mutant F-D3 were inoculated into the NIES-298 cultures of the exponential growth phase (at 15 d). (B) A genetic map indicating the transposon insertion site in mutant F-D3. (C) Growth of wild-type MAE1-K and mutant F-D3 in LB broth, BG-11 with glucose (0.1%) or with glucose and methionine (0.5 mM), and bleached culture. The bleached culture was prepared by filtering bleached NIES-298 culture at the end of the death phase. Optical density (600 nm) was measured after 24 h aerobic incubation (30°C). NG, no growth. (D) Mutant F-D3 that can still inactivate the bleaching property of bleached culture to NIES-298 culture.
FIG 7
FIG 7
Effects of methionine on the growth of NIES-298 cultures. Methionine (0.5 mM), together with the inoculation of wild-type MAE1-K or mutant F-D3, was supplemented into the NIES-298 cultures of the middle (15 d, A) and late (20 d, B) exponential growth phases.

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