Dataset Information

Emiliania Huxleyi exposed to 4 treatments of HHQ

ABSTRACT: Emiliania huxleyi: Cellular cascades induced by bacterial algicides Interactions between phytoplankton and bacteria play a central role in mediating oceanic biogeochemical cycling and microbial trophic structure in the ocean. The intricate relationships between these two domains of life are mediated via excreted molecules that facilitate communication and determine competitive outcomes. Yet, despite their predicted importance, identifying these secreted compounds and understanding their ecological significance has remained a challenge. Research in the Whalen Lab endeavors to (i) identify those bacterially-derived chemical signaling compounds (i.e. infochemicals) that mediate phytoplankton population dynamics, and (ii) determine the underlying physiological processes that contribute to phytoplankton tolerance or susceptibility to these compounds. Recently, the Whalen lab isolated an alkylquinolone-signaling molecule with known quorum sensing function from the globally distributed marine γ-proteobacteria, Pseudoalteromonas sp. capable of inducing species-specific phytoplankton mortality. This research was the first to suggest quorum sensing compounds have expanded and previously unrecognized ecological roles in regulating primary production and phytoplankton bloom dynamics. We are now investigating in how this alkylquinolone induces phytoplankton mortality via transcriptomic profiling and diagnostic biochemical analysis. Complementary to this transcriptomic examination, we will complete whole-cell proteomic approach to identify those phytoplankton proteins crucial in competitive interactions with bacterial infochemicals, but whose functions may not yet be known. With this proteomic approach in parallel to our transcriptomic investigation, we can establish a better understanding of the eukaryotic macromolecular targets and cellular cascades induced in response to bacterial algicides like alkylquinolones. With the knowledge gained from both approaches we can begin to address how these ?keystone molecules? influence population dynamics and community composition of phytoplankton and bacteria in field-based experiments with the goal of defining a new mechanistic framework for how bacterially derived signaling molecules influence biogeochemical cycles. D= DMSO - control treatment L= low 1 nm HHG additions M= medium 10 nm HHG additions H= high 100 nm HHG additions Each treatment had 4 biological replicates A-D

INSTRUMENT(S): Q Exactive

ORGANISM(S): Emiliania Huxleyi

TISSUE(S): Photosynthetic Cell, Whole Body

SUBMITTER: Brook Nunn

LAB HEAD: Brook L. Nunn

PROVIDER: PXD011560 | Pride | 2020-01-07

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	2018_Sept_28_Ehux.csv	Csv
	2018_Sept_28_Ehux_BLIB.blib	Other
	2018_Sept_28_Ehux_DA_36.mzML	Mzml
	2018_Sept_28_Ehux_DA_36.raw	Raw
	2018_Sept_28_Ehux_DB_21.raw	Raw

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Project description:Emiliania huxleyi: Cellular cascades induced by bacterial algicides Interactions between phytoplankton and bacteria play a central role in mediating oceanic biogeochemical cycling and microbial trophic structure in the ocean. The intricate relationships between these two domains of life are mediated via excreted molecules that facilitate communication and determine competitive outcomes. Yet, despite their predicted importance, identifying these secreted compounds and understanding their ecological significance has remained a challenge. Research in the Whalen Lab endeavors to (i) identify those bacterially-derived chemical signaling compounds (i.e. infochemicals) that mediate phytoplankton population dynamics, and (ii) determine the underlying physiological processes that contribute to phytoplankton tolerance or susceptibility to these compounds. Recently, the Whalen lab isolated an alkylquinolone-signaling molecule with known quorum sensing function from the globally distributed marine γ-proteobacteria, Pseudoalteromonas sp. capable of inducing species-specific phytoplankton mortality. This research was the first to suggest quorum sensing compounds have expanded and previously unrecognized ecological roles in regulating primary production and phytoplankton bloom dynamics. We are now investigating in how this alkylquinolone induces phytoplankton mortality via transcriptomic profiling and diagnostic biochemical analysis. Complementary to this transcriptomic examination, we will complete whole-cell proteomic approach to identify those phytoplankton proteins crucial in competitive interactions with bacterial infochemicals, but whose functions may not yet be known. With this proteomic approach in parallel to our transcriptomic investigation, we can establish a better understanding of the eukaryotic macromolecular targets and cellular cascades induced in response to bacterial algicides like alkylquinolones. With the knowledge gained from both approaches we can begin to address how these ?keystone molecules? influence population dynamics and community composition of phytoplankton and bacteria in field-based experiments with the goal of defining a new mechanistic framework for how bacterially derived signaling molecules influence biogeochemical cycles. D= DMSO - control treatment L= low 1 nm HHG additions M= medium 10 nm HHG additions H= high 100 nm HHG additions Each treatment had 4 biological replicates A-D

Project description:Phosphorus (P) is a critical driver of phytoplankton growth and ecosystem structure and function in the ocean. Diatoms are an abundant and widespread functional group of phytoplankton that are responsible for significant amounts of primary production in the ocean, however there has not been a comprehensive study of diatom physiological responses to P deficiency. Here, we coupled deep sequencing of transcript tags and quantitative proteomic analysis from the diatom Thalassiosira pseudonana grown under P-replete and P-deficient conditions. The reads (tags) were mapped to the T. pseudonana genome sequence, confirming expression of 91% of the modeled gene set. A total of 318 genes were differentially regulated with a false discovery rate of p<0.05. A total of 1264 proteins were detected, and of those 136 were differentially expressed with a false discovery rate of p<0.05. Significant changes in the abundance of transcripts and proteins were observed and these changes were coordinated for glycolysis, translation, and multiple biochemical responses to P deficiency. These data demonstrate that diatom P deficiency results in changes in cellular P allocation through polyphosphate production, increased P transport, a switch to utilization of dissolved organic P (DOP) through increased production of alkaline phosphatase metalloenzymes and a diesterase, and a remodeling of the cell surface through production of sulfolipids. Together, these findings reveal that T. pseudonana has evolved a sophisticated response to P deficiency involving multiple biochemical strategies that are likely critical to its ability to rapidly respond to variations in environmental P availability. T. pseudonana (Strain 1335 from the Provosoli-Guillard National Center for the Culture of Marine Phytoplankton (CCMP)) was grown in a modified f/2 medium made from Sargasso Sea water. Macronutrients and vitamin B12, biotin, and thiamine solutions were treated with prepared Chelex-100 resin to remove trace metal contaminants followed by trace-metal clean syringe sterilization to yield final nutrient concentrations of 882 μM NaNO3 and 106 μM Na2SiO3, and vitamin concentrations of 75 pM, 400 pM, and 60 nM, respectively. The Fe concentration was also modified from f/2 to 400 nM. All conditions were run in triplicate at 14 ºC, in constant light (120 µmol photons m-2 s-1). Cells were grown with f/2 phosphorus concentrations (P-replete; 36 µM PO4) and with low phosphorus concentrations (P-deficient; 0.4 µM PO4). Growth was monitored daily with cell counts. Duplicate P-replete treatments were pooled and harvested in mid log phase, and triplicate P-deficient treatments were pooled, and also quickly harvested onto 2 µm filters at the onset of P depletion. All RNA samples were snap frozen in liquid nitrogen. Replicate P-deficient cultures were refed to 36 µM phosphate at the onset of P depletion, and subsequently resumed growth. Additional growth studies were performed as described above substituting glycerolphosphate and adenosine monophosphate at 36 µM, for the phosphate in the medium.

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Emiliania Huxleyi exposed to 4 treatments of HHQ

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