skip to main content

Title: Vitamin B12 conveys a protective advantage to phycosphere-associated bacteria at high temperatures

Many marine microbes require vitamin B12(cobalamin) but are unable to synthesize it, necessitating reliance on other B12-producing microbes. Thus, phytoplankton and bacterioplankton community dynamics can partially depend on the production and release of a limiting resource by members of the same community. We tested the impact of temperature and B12availability on the growth of two bacterial taxa commonly associated with phytoplankton:Ruegeria pomeroyi, which produces B12and fulfills the B12requirements of some phytoplankton, andAlteromonas macleodii, which does not produce B12but also does not strictly require it for growth. For B12-producingR. pomeroyi, we further tested how temperature influences B12production and release. Access to B12significantly increased growth rates of both species at the highest temperatures tested (38 °C forR. pomeroyi, 40 °C forA. macleodii) andA. macleodiibiomass was significantly reduced when grown at high temperatures without B12, indicating that B12is protective at high temperatures. Moreover,R. pomeroyiproduced more B12at warmer temperatures but did not release detectable amounts of B12at any temperature tested. Results imply that increasing temperatures and more frequent marine heatwaves with climate change will influence microbial B12dynamics and could interrupt symbiotic resource sharing.

more » « less
Author(s) / Creator(s):
; ; ; ; ;
Publisher / Repository:
Nature Publishing Group
Date Published:
Journal Name:
ISME Communications
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. ABSTRACT Vitamin B 1 (thiamin) is a cofactor for critical enzymatic processes and is scarce in surface oceans. Several eukaryotic marine algal species thought to rely on exogenous thiamin are now known to grow equally well on the precursor 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP), including the haptophyte Emiliania huxleyi . Because the thiamin biosynthetic capacities of the diverse and ecologically important haptophyte lineage are otherwise unknown, we investigated the pathway in transcriptomes and two genomes from 30 species representing six taxonomic orders. HMP synthase is missing in data from all studied taxa, but the pathway is otherwise complete, with some enzymatic variations. Experiments on axenic species from three orders demonstrated that equivalent growth rates were supported by 1 µM HMP or thiamin amendment. Cellular thiamin quotas were quantified in the oceanic phytoplankter E. huxleyi using the thiochrome assay. E. huxleyi exhibited luxury storage in standard algal medium [(1.16 ± 0.18) × 10 −6  pmol thiamin cell −1 ], whereas quotas in cultures grown under more environmentally relevant thiamin and HMP supplies [(2.22 ± 0.07) × 10 −7 or (1.58 ± 0.14) × 10 −7  pmol thiamin cell −1 , respectively] were significantly lower than luxury values and prior estimates. HMP and its salvage-related analog 4-amino-5-aminomethyl-2-methylpyrimidine (AmMP) supported higher growth than thiamin under environmentally relevant supply levels. These compounds also sustained growth of the stramenopile alga Pelagomonas calceolata . Together with identification of a salvage protein subfamily (TENA_E) in multiple phytoplankton, the results indicate that salvaged AmMP and exogenously acquired HMP are used by several groups for thiamin production. Our studies highlight the potential importance of thiamin pathway intermediates and their analogs in shaping phytoplankton community structure. IMPORTANCE The concept that vitamin B 1 (thiamin) availability in seawater controls the productivity and structure of eukaryotic phytoplankton communities has been discussed for half a century. We examined B 1 biosynthesis and salvage pathways in diverse phytoplankton species. These comparative genomic analyses as well as experiments show that phytoplankton thought to require exogenous B 1 not only utilize intermediate compounds to meet this need but also exhibit stronger growth on these compounds than on thiamin. Furthermore, oceanic phytoplankton have lower cellular thiamin quotas than previously reported, and salvage of intermediate compounds is likely a key mechanism for meeting B 1 requirements under environmentally relevant scenarios. Thus, several lines of evidence now suggest that availability of specific precursor molecules could be more important in structuring phytoplankton communities than the vitamin itself. This understanding of preferential compound utilization and thiamin quotas will improve biogeochemical model parameterization and highlights interaction networks among ocean microbes. 
    more » « less
  2. Abstract

    A complex interplay of environmental variables impacts phytoplankton community composition and physiology. Temperature and nutrient availability are two principal factors driving phytoplankton growth and composition, but are often investigated independently and on individual species in the laboratory. To assess the individual and interactive effects of temperature and nutrient concentration on phytoplankton community composition and physiology, we altered both the thermal and nutrient conditions of a cold‐adapted spring phytoplankton community in Narragansett Bay, Rhode Island, when surface temperature was 2.6°C and chlorophyll > 9 μg L−1. Water was incubated in triplicate at −0.5°C, 2.6°C, and 6°C for 10 d. At each temperature, treatments included both nutrient amendments (N, P, Si addition) and controls (no macronutrients added). The interactive effects of temperature and resource availability altered phytoplankton growth and community structure. Nutrient amendments resulted in species sorting and communities dominated by larger species. Under replete nutrients, warming tripled phytoplankton growth rates, but under in situ nutrient conditions, increased temperature acted antagonistically, reducing growth rates by as much as 33%, suggesting communities became nutrient limited. The temperature–nutrient interplay shifted the relative proportions of each species within the phytoplankton community, resulting in more silica rich cells at decreasing temperatures, irrespective of nutrients, and C : N that varied based on resource availability, with nutrient limitation inducing a 47% increase in C : N at increasing temperatures. Our results illustrate how the temperature–nutrient interplay can alter phytoplankton community dynamics, with changes in temperature amplifying or exacerbating the nutrient effect with implications for higher trophic levels and carbon flux.

    more » « less
  3. To assess protistan grazing impact and temperature sensitivity on plankton population dynamics, we measured bulk and species-specific phytoplankton growth and herbivorous protist grazing rates in Disko Bay, West Greenland in April-May 2011. Rate estimates were made at three different temperatures in situ (0 °C), +3 °C and +6 °C over ambient. In situ Chlorophyll a (Chl a ) doubled during the observation period to ∼12  µg Chl a L −1 , with 60–97% of Chl a in the >20 µm size-fraction dominated by the diatom genus Chaetoceros. Herbivorous dinoflagellates comprised 60–80% of microplankton grazer biomass. At in situ temperatures, phytoplankton growth or grazing by herbivorous predators <200 µm was not measurable until 11 days after observations commenced. Thereafter, phytoplankton growth was on average 0.25 d −1 . Phytoplankton mortality due to herbivorous grazing was only measured on three occasions but the magnitude was substantial, up to 0.58 d −1 . Grazing of this magnitude removed ∼100% of primary production. In short-term temperature-shift incubation experiments, phytoplankton growth rate increased significantly (20%) at elevated temperatures. In contrast, herbivorous protist grazing and species-specific growth rates decreased significantly (50%) at +6 °C. This differential response in phytoplankton and herbivores to temperature increases resulted in a decrease of primary production removed with increasing temperature. Phaeocystis spp. abundance was negatively correlated with bulk grazing rate. Growth and grazing rates were variable but showed no evidence of an inherent, low temperature limitation. Herbivorous protist growth rates in this study and in a literature review were comparable to rates from temperate waters. Thus, an inherent physiological inhibition of protistan growth or grazing rates in polar waters is not supported by the data. The large variability between lack of grazing and high rates of primary production removal observed here and confirmed in the literature for polar waters implies larger amplitude fluctuations in phytoplankton biomass than slower, steady grazing losses of primary production. 
    more » « less
  4. Abstract

    The heterotrophic marine bacterium,Ruegeria pomeroyi, was experimentally cultured under environmentally realistic carbon conditions and with a tracer-level addition of13C-labeled leucine to track bacterial protein biosynthesis through growth phases. A combination of methods allowed observation of real-time bacterial protein production to understand metabolic priorities through the different growth phases. Over 2000 proteins were identified in each experimental culture from exponential and stationary growth phases. Within two hours of the13C-labeled leucine addition,R.pomeroyisignificantly assimilated the newly encountered substrate into new proteins. This dataset provides a fundamental baseline for understanding growth phase differences in molecular physiology of a cosmopolitan marine bacterium.

    more » « less
  5. ABSTRACT There is a growing appreciation within animal and plant physiology that the reactive oxygen species (ROS) superoxide is not only detrimental but also essential for life. Yet, despite widespread production of extracellular superoxide by healthy bacteria and phytoplankton, this molecule remains associated with stress and death. Here, we quantify extracellular superoxide production by seven ecologically diverse bacteria within the Roseobacter clade and specifically target the link between extracellular superoxide and physiology for two species. We reveal for all species a strong inverse relationship between cell-normalized superoxide production rates and cell number. For exponentially growing cells of Ruegeria pomeroyi DSS-3 and Roseobacter sp. strain AzwK-3b, we show that superoxide levels are regulated in response to cell density through rapid modulation of gross production and not decay. Over a life cycle of batch cultures, extracellular superoxide levels are tightly regulated through a balance of both production and decay processes allowing for nearly constant levels of superoxide during active growth and minimal levels upon entering stationary phase. Further, removal of superoxide through the addition of exogenous superoxide dismutase during growth leads to significant growth inhibition. Overall, these results point to tight regulation of extracellular superoxide in representative members of the Roseobacter clade, consistent with a role for superoxide in growth regulation as widely acknowledged in fungal, animal, and plant physiology. IMPORTANCE Formation of reactive oxygen species (ROS) through partial reduction of molecular oxygen is widely associated with stress within microbial and marine systems. Nevertheless, widespread observations of the production of the ROS superoxide by healthy and actively growing marine bacteria and phytoplankton call into question the role of superoxide in the health and physiology of marine microbes. Here, we show that superoxide is produced by several marine bacteria within the widespread and abundant Roseobacter clade. Superoxide levels outside the cell are controlled via a tightly regulated balance of production and decay processes in response to cell density and life stage in batch culture. Removal of extracellular superoxide leads to substantial growth inhibition. These findings point to an essential role for superoxide in the health and growth of this ubiquitous group of microbes, and likely beyond. 
    more » « less