In this study we established the B1 and B12 vitamin requirement of the dinoflagellate Lingulodinium polyedrum and the vitamin supply by its associated bacterial community. In previous field studies the B1 and B12 demand of this species was suggested but not experimentally verified. When the axenic vitamin un-supplemented culture (B-ns) of L. polyedrum was inoculated with a coastal bacterial community, the dinoflagellate’s vitamin growth limitation was overcome, reaching the same growth rates as the culture growing in vitamin B1B7B12-supplemented (B-s) medium. Measured B12 concentrations in the B-s and B-ns cultures were both higher than typical coastal concentrations and B12 in the B-s culture was higher than in the B-ns culture. In both B-s and B-ns cultures, the probability of dinoflagellate cells having bacteria attached to the cell surface was similar and in both cultures an average of six bacteria were attached to each dinoflagellate cell. In the B-ns culture the free bacterial community showed significantly higher cell abundance suggesting that unattached bacteria supplied the vitamins. The fluorescence in situ hybridization (FISH) protocol allowed the quantification and identification of three bacterial groups in the same samples of the free and attached epibiotic bacteria for both treatments. The relative composition of these groups was not significantly different and was dominated by Alphaproteobacteria (>89%). To complement the FISH counts, 16S rDNA sequencing targeting the V3–V4 regions was performed using Illumina-MiSeq technology. For both vitamin amendments, the dominant group found was Alphaproteobacteria similar to FISH, but the percentage of Alphaproteobacteria varied between 50 and 95%. Alphaproteobacteria were mainly represented by Marivita sp., a member of the Roseobacter clade, followed by the Gammaproteobacterium Marinobacter flavimaris. Our results show that L. polyedrum is a B1 and B12 auxotroph, and acquire both vitamins from the associated bacterial community in sufficient quantity to sustain the maximum growth rate.
Oxygen minimum zones (OMZs) are important sources of CO 2 to the atmosphere when physical forces bring subsurface water with high dissolved inorganic carbon (DIC) to the surface. This study examines, for the first time, the influence of the OMZ of the coastal North Eastern Tropical Pacific off Mexico on surface CO 2 fluxes. We use variations in the oxycline depth and subsurface water masses to discern physical oceanographic influences. During two cruises, in November 2009 and August 2010, DIC and total alkalinity (TA) measurements were used to estimate pCO 2 and air-sea CO 2 fluxes. At the OMZ layer, Subtropical Subsurface Water (StSsW) was found to have high pCO 2 values (1290 6 70 latm). Due to strong vertical stratification, however, the relationship between DpCO 2 at the air-sea interface and the oxycline/StSsW upper limit depth was weak. During November, the region was a weak source of CO 2 to the atmosphere (up to 2.5 mmol C m ) prevented subsurface mixing of water from the OMZ to the upper layer; particularly in November 2009 which was during an El Niño event. Results suggest that advection of surface water masses, reinforced by strong vertical stratification, controlled surface pCO 2 , and air-sea CO 2 fluxes.
Spectra1 absorption of phytoplankton from cultures and natural samples was measured on filters with various optical setups including collimated and diffuse irradiation and measurements ofwetted filters within an integrating sphere. In suspensions within an integrating sphere, specific absorption coefficients for laboratory cultures varied by a factor of only two.Measurements on filters yielded values dependent on filter load. Specific absorption coefficients derived from measurements of sample filters were considerably higher than values obtained from suspensions in an integrating sphere due to increased diffuseness of irradiance and to pathlength amplification by filter-particle and particle-particle interactions. Measured absorption of phytoplankton in the blue can be increased greatly by absorption of detritus, evident from absorption spectra of depigmented samples on filter. After subtracting detrital absorption, absorption spectra of phytoplankton are qualitatively similar to the corresponding quantum-corrected fluorescence excitation spectra. The detritus-corrected ratio of absorption at 440 vs. 675 nm shows average values between 1 and 1.5.Comparison with published values shows that specific in vivo absorption coefficients of phytoplankton arc mostly overestimated as a result of the methodology applied. In the blue region of the spectrum, overestimation of phytoplankton absorption in field samples is possible if detrital absorption is neglected.
Prokaryote respiration is expected to be responsible for more than half of the community respiration in the ocean, but the lack of a practical method to measure the rate of prokaryote respiration in the open ocean resulted in very few published data leaving the role of organotrophic prokaryotes open to debate. Oxygen consumption rates of oceanic prokaryotes measured with current methods may be biased due to pre-incubation size filtration and long incubation times both of which can change the physiological and taxonomic profile of the sample during the incubation period. In vivo INT reduction has been used in terrestrial samples to estimate respiration rates, and recently, the method was introduced and applied in aquatic ecology. We measured oxygen consumption rates and in vivo INT reduction to formazan in cultures of marine bacterioplankton communities, Vibrio harveyi and the eukaryote Isochrysis galbana. For prokaryotes, we observed a decrease in oxygen consumption rates with increasing INT concentrations between 0.05 and 1 mM. Time series after 0.5 mM INT addition to prokaryote samples showed a burst of in vivo INT reduction to formazan and a rapid decline of oxygen consumption rates to zero within less than an hour. Our data for non-axenic eukaryote cultures suggest poisoning of the eukaryote. Prokaryotes are clearly poisoned by INT on time scales of less than 1 h, invalidating the interpretation of in vivo INT reduction to formazan as a proxy for oxygen consumption rates.
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