The distinctive level of interaction between carbon and nitrogen metabolisms in the leaves of submerged macrophytes plays a key role in ammonium detoxification.

Possible ammonium detoxification mechanisms have been proposed recently, on submerged macrophytes, evidently illustrating that glutamate dehydrogenase (GDH) plays a greater role in ammonium detoxification compared to the primary glutamine synthetase/glutamate oxaloacetate transaminase (GS/GOGAT) pathway. In the current investigation, we cultured three submerged macrophytes to extreme concentrations of [NH4+-N] of up to 50 mg/L with the aim of clarifying the interaction between carbon and nitrogen metabolisms. The activities of carboxylation enzymes pyruvate orthophosphate dikinase (PPDK) and phosphoenolpyruvate carboxylase (PEPC), in lieu of Rubisco, increased almost two-fold for ammonium tolerant species P. maackianus and M. spicatum, compared with the sensitive species P. lucens. While these enzymes are well known for their central role in CO2 fixation, their inference in conferring resistance to ammonium stress has not been well elucidated before. In this study, we demonstrate that the overproduction of PEPC and PPDK led to improved photosynthesis, better ammonium assimilation and overall ammonium detoxification in M. spicatum and P. maackianus. These findings propose likelihood for the existence of a complementary ammonium detoxification pathway that targets carbon metabolism, thus, presenting a relatively efficient linkage between nitrogen and carbon metabolisms and identify candidate species for practical restoration of fresh water resources.

Transcriptome Analysis of Macrophytes' Myriophyllum spicatum Response to Ammonium Nitrogen Stress Using the Whole Plant Individual.

Ammonium toxicity in macrophytes reduces growth and development due to a disrupted metabolism and high carbon requirements for internal ammonium detoxification. To provide more molecular support for ammonium detoxification in the above-ground and below-ground parts of Myriophyllum spicatum, we separated (using hermetic bags) the aqueous medium surrounding the below-ground from that surrounding the above-ground and explored the genes in these two regions. The results showed an upregulation of asparagine synthetase genes under high ammonium concentrations. Furthermore, the transcriptional down and/or upregulation of other genes involved in nitrogen metabolism, including glutamate dehydrogenase, ammonium transporter, and aspartate aminotransferase in above-ground and below-ground parts were crucial for ammonium homeostasis under high ammonium concentrations. The results suggest that, apart from the primary pathway and alternative pathway, the asparagine metabolic pathway plays a crucial role in ammonium detoxification in macrophytes. Therefore, the complex genetic regulatory network in M. spicatum contributes to its ammonium tolerance, and the above-ground part is the most important in ammonium detoxification. Nevertheless, there is a need to incorporate an open-field experimental setup for a conclusive picture of nitrogen dynamics, toxicity, and the molecular response of M. spicatum in the natural environment.

The Above-Ground Part of Submerged Macrophytes Plays an Important Role in Ammonium Utilization.

As a paradoxical nutrient in water ecosystems, ammonium can promote plants growth under moderate concentration, but excess of it causes phytotoxic effects. Previous research has revealed that glutamate dehydrogenase in the above-ground part of submerged macrophytes plays an important role in ammonium detoxification. However, the strategies of ammonium utilization at the whole plant level of submerged macrophytes are still unclear and the role of the above-ground part in nutrient utilization has not been clearly elucidated in previous studies, hence, directly influencing the application of previous theory to practice. In the present research, we combined the methods of isotopic labeling and enzyme estimation to investigate strategies of ammonium utilization by the submerged macrophytes. The results showed that when [NH4 +-N] was 50 mg L-1, 15N taken up through the above-ground parts was 13.24 and 17.52 mg g-1 DW, while that of the below-ground parts was 4.24 and 8.54 mg g-1 DW in Potamogeton lucens and Myriophyllum spicatum, respectively. The ratios of 15N acropetal translocation to uptake were 25.75 and 35.69%, while those of basipetal translocation to uptake were 1.93 and 4.09% in P. lucens and M. spicatum, respectively. Our results indicated that the above-ground part was not only the main part for ammonium uptake, but also the major pool of exogenous ammonium. Besides, the dose-response curve of GDH (increased by 20.9 and 50.2% under 15 and 50 mg L-1 [NH4 +-N], respectively) exhibited by the above-ground parts of M. spicatum indicates that it is the main site for ammonium assimilation of the tolerant species. This study identifies the ammonium utilization strategy of submerged macrophytes and reveals the important role of the above-ground part in nutrient utilization providing new insight into the researches of nutrient utilization by plants and theoretical supports for water restoration by phytoremediation.

Exploring the ammonium detoxification mechanism of young and mature leaves of the macrophyte Potamogeton lucens.

Toxicity in aquatic plants, caused by excess ammonium in the environment, is an important ecological problem and active research topic. Recent studies showed the importance of the enzyme Glutamate Dehydrogenase (GDH) in detoxifying ammonium. However, these results mainly derived from species comparisons, hence some mechanisms may have been obscured due to species differences. Our recent finding that young leaves of Potamogeton lucens were less sensitive to ammonium enrichment, than mature leaves allowed us to study ammonium detoxification within a species. We found that, unlike mature leaves, ammonium-tolerant young leaves of P. lucens could assimilate ammonium mainly through GDH. There was a 38% increase of NADH-dependent GDH in 50 mg/L ammonium concentration compared with 0.1 mg/L. Therefore, this study confirms the hypothesis that the GDH pathway plays a major role in the detoxification of ammonium in freshwater macrophytes.

Prevalence of antibodies to Brucella species in commercial raw bovine milk in Southwestern Uganda.

OBJECTIVES: The purpose and objective of this research was to explore the prevalence of antibodies against Brucella species in raw milk samples collected in Southwestern Uganda, one of the biggest milk producing regions in the Country. We hypothesized that there is a high level of antibodies in milk samples from this region. This builds more evidence to other studies in the region on the level contamination of raw milk. RESULTS: A total of 185 raw milk samples, collected from dairy farms and factories in southwestern region, were tested for antibodies to Brucella spp. using the milk ring test (MRT) and indirect Enzyme-Linked Immunosorbent Assay (i-ELISA).We found a prevalence of 26.5% (49/185) by the two methods. This is related to previous reports in the region and adds more evidence on the need for further investigations to confirm the source of these antibodies and their relationship with disease in milk producing animals.