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. 2022 Nov 22;88(22):e0161922.
doi: 10.1128/aem.01619-22. Epub 2022 Oct 27.

Dynamic Diatom-Bacteria Consortia in Synthetic Plankton Communities

Affiliations

Dynamic Diatom-Bacteria Consortia in Synthetic Plankton Communities

Yun Deng et al. Appl Environ Microbiol. .

Abstract

Microalgae that form phytoplankton live and die in a complex microbial consortium in which they co-exist with bacteria and other microorganisms. The dynamics of species succession in the plankton depends on the interplay of these partners. Bacteria utilize substrates produced by the phototrophic algae, while algal growth can be supported by bacterial exudates. Bacteria might also use chemical mediators with algicidal properties to attack algae. To elucidate whether specific bacteria play universal or context-specific roles in the interaction with phytoplankton, we investigated the effect of cocultured bacteria on the growth of 8 microalgae. An interaction matrix revealed that the function of a given bacterium is highly dependent on the cocultured partner. We observed no universally algicidal or universally growth-promoting bacteria. The activity of bacteria can even change during the aging of an algal culture from inhibitory to stimulatory or vice versa. We further established a synthetic phytoplankton/bacteria community with the centric diatom, Coscinodiscus radiatus, and 4 phylogenetically distinctive bacterial isolates, Mameliella sp., Roseovarius sp., Croceibacter sp., and Marinobacter sp. Supported by a Lotka-Volterra model, we show that interactions within the consortium are specific and that the sum of the pairwise interactions can explain algal and bacterial growth in the community. No synergistic effects between bacteria in the presence of the diatom was observed. Our survey documents highly species-specific interactions that are dependent on algal fitness, bacterial metabolism, and community composition. This species specificity may underly the high complexity of the multi-species plankton communities observed in nature. IMPORTANCE The marine food web is fueled by phototrophic phytoplankton. These algae are central primary producers responsible for the fixation of ca. 40% of the global CO2. Phytoplankton always co-occur with a diverse bacterial community in nature. This diversity suggests the existence of ecological niches for the associated bacteria. We show that the interaction between algae and bacteria is highly species-specific. Furthermore, both, the fitness stage of the algae and the community composition are relevant in determining the effect of bacteria on algal growth. We conclude that bacteria should not be sorted into algicidal or growth supporting categories; instead, a context-specific function of the bacteria in the plankton must be considered. This functional diversity of single players within a consortium may underly the observed diversity in the plankton.

Keywords: bacteria; consortia; diatoms; host-cell interactions; interactions; microalgae; microbiome; phytoplankton.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

FIG 1
FIG 1
Pairwise co-cultivation to identify interaction patterns between bacteria and diatoms. (a) Cell morphology of the 8 diatoms used in this study. Clockwise from the top left: C. didymus, C. socialis, T. weissflogii, T. nordenskioeldii, C. wailesii, S. hyperborea, C. granii, and C. radiatus. (b) Heatmaps summarize the effects of bacteria on diatoms in their early (before reaching the chlorophyll maximum) and late (after reaching the chlorophyll maximum) growth phase. The workflow for plotting the heatmaps is described in Fig. S2. Average of n = 3 replicates is plotted, growth curves for each pairwise interaction are given in Fig. S3-S10. *Data were normalized by the average cell densities determined for all cocultures with different bacterial inoculations. Chla FC refers to Chlorophyll a fold change.
FIG 2
FIG 2
Cell counts in the co-cultivation of the diatom C. radiatus with the individual bacterial species, Roseovarius Rose1, Croceibacter Crocei1, Marinobacter CS1 and 4 bacteria in combination. (a) Scheme for the coculture experiment design, monoculture C. radiatus (upper) and C. radiatus-bacteria coculture (lower). Live cell counts of C. radiatus in coculture with the bacteria CS4 (b), Rose1(c), Crocei1(d), CS1(e) and in controls without added bacteria (orange). The effect of the coculture of the synthetic community with four bacteria combination is shown in (f). Bars derive from SE of three biological replicates. Unpaired t test, ** indicates P < 0.01, * indicates P < 0.05.
FIG 3
FIG 3
Bacterial quantification in the co-cultivation of the diatom C. radiatus with the individual bacteria, Roseovarius Rose1, Croceibacter Crocei1, Marinobacter CS1 and four bacteria in combination. (a) Scheme for the coculture experiment design, bacterial monoculture (upper) and C. radiatus-bacteria coculture (lower). Bacterial density was quantified by qPCR when initially inoculated with bacterial strains CS4 (b), Rose1(c), Crocei1(d), CS1(e) and 4 bacteria in combination (f). Controls without added diatoms are coded with gray. Bacterial cells mL−1 are plotted on a logarithmic scale. Bars derive from SE of three biological replicates. Unpaired t test, **** indicates P < 0.001, *** indicates P < 0.005, ** indicates P < 0.01, * indicates P < 0.05.
FIG 4
FIG 4
(a) Prediction of the mathematical Lotka-Volterra model for algal growth in the presence of all four bacteria. The black line shows the model prediction using parameters obtained by fitting to the data from isolated cultures and pairwise cocultures. The model agrees well with the experimental data (blue line). For comparison, we plotted also the data for diatom growth in isolation (orange line) and the corresponding model fit (red line). (b) Illustration of the inter-species interactions obtained from the model fit of the pairwise coculture data. Neutral, stimulatory, and inhibitory interactions are indicated by blue circle-headed, red flat and green pointed arrows, respectively. The strongest interaction detected by the model is stimulation of Crocei1 by C. radiatus.
FIG 5
FIG 5
The requirement of the four bacteria (a) CS4, (b) Rose1, (c) Crocei1, and (d) CS1 for nutrients in a minimal medium. The optical density OD600 was recorded and used for representing the bacterial growth at an early time point, where first growth was detected by visual inspection (black column) and 24 h later (gray column). Error bars represent the standard deviation of three biological replicates. The statistical analyses of the multiple comparisons are provided in the Table S6.

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