Harmful algal blooms (HABs) can be detrimental to ecosystems, human health, and economies. The low levels of phosphorus remaining in the effluent of municipal wastewater treatment plants can contribute to HAB formation. To achieve more complete phosphorus removal, an effluent treatment method has been proposed that uses phosphorus-deprived, filamentous microalgae to quickly assimilate soluble phosphorus to low concentrations. This study investigated two parameters that influence the feasibility of such a system: (1) the biomass growth productivity of algal cultures during the phosphorus deprivation period and (2) the correlation between the duration of this period and the phosphorus uptake rate by the biomass when contacted with the water to be treated. A single strain of filamentous algae, Tribonema minus, was used. Two experiments lasting 8-9 days compared the biomass productivity of cultures of T. minus grown in phosphorus-replete and -deplete media. While no significant difference in productivity was observed between treatments, further studies should be done to confirm this finding. In addition, 39 uptake contact experiments were conducted. The soluble phosphorus uptake rate was measured for algae deprived of phosphorus for 0 to 12 days of growth. The highest observed uptake rate was 3.83 mg P/g VSS-h, during the first three hours of contact, by biomass that had been phosphorus-deprived for 12 days. The correlation between phosphorus deprivation duration and 3-h uptake rate was 0.34 mg P/g VSS-h per day of deprivation (R2 = 0.81). Additional development efforts seem justified based on these results.

Symbiotic nitrogen (N) fixation (SNF) occurs in specialized organs called nodules after successful interactions between legume hosts and rhizobia. Within nodule cells, N-fixing rhizobia are surrounded by plant-derived symbiosome membranes, through which the exchange of nutrients and ammonium occurs between bacteria and the host legume. Phosphorus (P) is an essential macronutrient, and N2-fixing legumes have a higher requirement for P than legumes grown on mineral N. First, I investigated the impact of P deprivation on wild-type Medicago truncatula plants. My observations that plants had impaired SNF activity, reduced growth, and accumulated less phosphate in P-deficient tissues (leaves, roots and nodules) is consistent with those of similar previous studies. Galactolipids decreased with increase in phospholipids in all P-starved organs. Matrix-assisted laser desorption/ionization–mass spectrometry imaging (MALDI-MSI) of phosphatidylcholine (PC) species in nodules showed that under low P environments distributions of some PC species changed, indicating that membrane lipid remodeling during P stress is not uniform across the nodule. Secondly, a metabolomics study was carried out to test the alterations in the metabolic profile of the nodules in P-stress. GC-MS based untargeted metabolomics showed increased levels of amino acids and sugars and decline in amounts of organic acids in P deprived nodules. Subsequently, LC-MS/MS was used to quantify these compounds including phosphorylated metabolites in whole plant. My findings showed strong drop in levels of organic acids and phosphorylated compounds in P deprived leaves with moderate reduction in P deprived roots and nodules. Moreover, sugars and amino acids were elevated in whole plant under P deprivation. Finally, the last project of my thesis involved studying the response of PDIL3 (Phosphate Deficiency-Induced LncRNA-3) a long non-coding RNA (lncRNA) mutant under severe P stress. PDIL3 is known to regulate Pi-deficiency signaling and transport in M. truncatula (Wang et al., 2017). My results confirmed that in P starvation, pdil3 plants showed better shoot growth, accumulated more phosphate in shoots, had impaired SNF and less rhizobial occupancy in nodules than WT. Subsequently, MALDI–MS imaging was used to spatially map and compare the distribution of phosphatidylcholine (PC) species in nodules of pdil3 and WT in P-replete and P-depleted conditions. Several PC species showed changes in distributions in pdil3 nodules compared to WT in both P sufficient and P deprived conditions. These data suggest that PDIL3's role is not just suppression of the Pi transporter, but it may also influence P partitioning between shoots and nodulated roots, meriting further investigation.