Endogenous Production and Vibrational Analysis of Heavy-Isotope-Labeled Peptides from Cyanobacteria.

Stable isotope labeling is an extremely useful tool for characterizing the structure, tracing the metabolism, and imaging the distribution of natural products in living organisms using mass-sensitive measurement techniques. In this study, a cyanobacterium was cultured in 15 N/13 C-enriched media to endogenously produce labeled, bioactive oligopeptides. The extent of heavy isotope incorporation in these peptides was determined with LC-MS, while the overall extent of heavy isotope incorporation in whole cells was studied with nanoSIMS and AFM-IR. Up to 98 % heavy isotope incorporation was observed in labeled cells. Three of the most abundant peptides, microcystin-LR (MCLR), cyanopeptolin-A (CYPA), and aerucyclamide-A (ACAA), were isolated and further studied with Raman and FTIR spectroscopies and DFT calculations. This revealed several IR and Raman active vibrations associated with functional groups not common in ribosomal peptides, like diene, ester, thiazole, thiazoline, and oxazoline groups, which could be suitable for future vibrational imaging studies. More broadly, this study outlines a simple and relatively inexpensive method for producing heavy-labeled natural products. Manipulating the bacterial culture conditions by the addition of specific types and amounts of heavy-labeled nutrients provides an efficient means of producing heavy-labeled natural products for mass-sensitive imaging studies.

Phosphorus retention within a relic agricultural ditch in a constructed wetland.

Stormwater treatment areas (STAs) are constructed wetlands established to capture phosphorus (P) from agricultural runoff before reaching the Florida Everglades. Retained P is primarily stored in wetland soils and sediments generated through a collection of interrelated physical, chemical, and biological processes. The amount of P and other elements (Al, Ca, Cu, Fe, Mg, Mn, Pb, and Zn) retained in the flocculent (floc) and recently accreted soil (RAS) horizons from a relic agricultural ditch within Cell 4S of STA-1E were compared with the surrounding marsh soils (upstream and downstream sites of the ditch). The amount of P retained in the ditch was significantly greater than the surrounding marsh soils and for all the elements in the floc horizon and five of the nine elements in the RAS horizon, suggesting that different processes or process rates influenced accumulation. Phosphorus species in the floc and RAS sediment horizons were identified and quantified using 31 P nuclear magnetic resonance (NMR) spectroscopy and total P determined by microwave plasma-optical emission spectroscopy. In general, P forms were dominated by orthophosphate, sugar phosphates, nucleotides, DNA, and pyrophosphate, with varying relative abundances of species. Total P concentration significantly decreased from upstream to downstream of the ditch by an average of 28 and 35% for floc and RAS soils, respectively. The relatively high P accrual rate within the ditch suggested that relic ditches perpendicular to flow could reduce P transport to downstream soils and sediments and, in turn, help maintain low P levels in overlying water.

The role of rice (Oryza sativa L.) in sequestering phosphorus compounds and trace elements: Speciation and dynamics.

In southern Florida, the sequestering of nutrients through the cultivation of rice (Oryza sativa L.) in alternation with sugarcane (Saccharum spp.) crops is an essential step in minimizing downstream eutrophication of the Florida Everglades. Phosphorus (P) is known to be the leading cause of this eutrophication; however, the cultivation/harvesting of rice effectively reduces P and additional macro and micro-nutrients from agrarian soil and runoff through plant uptake. In this study, soil, water, sugarcane, and rice plants at two different stages (flooded and vegetative) were analyzed for twelve different elements (Al, As, Co, Cr, Cu, Fe, Ni, Zn, Ca, Mn, Mg, and P) by Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES). 31P Nuclear Magnetic Resonance (NMR) spectra of the rice plants confirmed ten different P compounds being transported and/or transformed throughout the entirety of the sugarcane and rice plants. On average, dried rice plants contained 1677 ± 14 mg-P, of which 1277 ± 3.0 mg-P was in the panicle at the vegetative stage. Harvesting of the rice panicle has the potential to remove about 14.7 kg-P/ha for the top 10 cm of the soil. This present study demonstrates that in rotations with sugarcane crops and with no added P fertilizer, rice cultivation can reduce considerable amounts of P that would otherwise leach into the Greater Everglades from the Everglades Agricultural Area.

Brevetoxin (PbTx-2) influences the redox status and NPQ of Karenia brevis by way of thioredoxin reductase.

The Florida red tide dinoflagellate, Karenia brevis, is the major harmful algal bloom dinoflagellate of the Gulf of Mexico and plays a destructive role in the region. Blooms of K. brevis can produce brevetoxins: ladder-shaped polyether (LSP) compounds, which can lead to adverse human health effects, such as reduced respiratory function through inhalation exposure, or neurotoxic shellfish poisoning through consumption of contaminated shellfish. The endogenous role of the brevetoxins remains uncertain. Recent work has shown that some forms of NADPH dependent thioredoxin reductase (NTR) are inhibited by brevetoxin-2 (PbTx-2). The study presented herein reveals that high toxin and low toxin K. brevis, which have a ten-fold difference in toxin content, also show a significant difference in their ability, not only to produce brevetoxin, but also in their cellular redox status and distribution of xanthophyll cycle pigments. These differences are likely due to the inhibition of NTR by brevetoxin. The work could shed light on the physiological role that brevetoxin fills for K. brevis.