The basic Leucine zipper transcription factor ABSCISIC ACID INSENSITIVE5 (ABI5) is a key regulator of abscisic acid (ABA)-mediated seed germination and postgermination seedling growth. While a family of SUCROSE NONFERMENTING1-related protein kinase2s (SnRK2s) is responsible for ABA-induced phosphorylation and stabilization of ABI5, the phosphatase(s) responsible for dephosphorylating ABI5 is still unknown. Here, we demonstrate that mutations in FyPP1 (for Phytochromeassociated serine/threonine protein phosphatase1) and FyPP3, two homologous genes encoding the catalytic subunits of Ser/ Thr PROTEIN PHOSPHATASE6 (PP6), cause an ABA hypersensitive phenotype in Arabidopsis thaliana, including ABA-mediated inhibition of seed germination and seedling growth. Conversely, overexpression of FyPP causes reduced sensitivity to ABA. The ABA hypersensitive phenotype of FyPP loss-of-function mutants is ABI5 dependent, and the amount of phosphorylated and total ABI5 proteins inversely correlates with the levels of FyPP proteins. Moreover, FyPP proteins physically interact with ABI5 in vitro and in vivo, and the strength of the interaction depends on the ABI5 phosphorylation status. In vitro phosphorylation assays show that FyPP proteins directly dephosphorylate ABI5. Furthermore, genetic and biochemical assays show that FyPP proteins act antagonistically with SnRK2 kinases to regulate ABI5 phosphorylation and ABA responses. Thus, Arabidopsis PP6 phosphatase regulates ABA signaling through dephosphorylation and destabilization of ABI5.
An activated sludge model for greenhouse gases no. 1 was calibrated with data from a wastewater treatment plant (WWTP) without control systems and validated with data from three similar plants equipped with control systems. Special about the calibration/validation approach adopted in this paper is that the data are obtained from simulations with a mathematical model that is widely accepted to describe effluent quality and operating costs of actual WWTPs, the Benchmark Simulation Model No. 2 (BSM2). The calibration also aimed at fitting the model to typical observed nitrous oxide (N₂O) emission data, i.e., a yearly average of 0.5% of the influent total nitrogen load emitted as N₂O-N. Model validation was performed by challenging the model in configurations with different control strategies. The kinetic term describing the dissolved oxygen effect on the denitrification by ammonia-oxidizing bacteria (AOB) was modified into a Haldane term. Both original and Haldane-modified models passed calibration and validation. Even though their yearly averaged values were similar, the two models presented different dynamic N₂O emissions under cold temperature conditions and control. Therefore, data collected in such situations can potentially permit model discrimination. Observed seasonal trends in N₂O emissions are simulated well with both original and Haldane-modified models. A mechanistic explanation based on the temperature-dependent interaction between heterotrophic and autotrophic N₂O pathways was provided. Finally, while adding the AOB denitrification pathway to a model with only heterotrophic N₂O production showed little impact on effluent quality and operating cost criteria, it clearly affected N2O emission productions.
Five activated sludge models describing N2O production by ammonium oxidising bacteria (AOB) were compared to four different long-term process data sets. Each model considers one of the two known N2O production pathways by AOB, namely the AOB denitrification pathway and the hydroxylamine oxidation pathway, with specific kinetic expressions. Satisfactory calibration could be obtained in most cases, but none of the models was able to describe all the N2O data obtained in the different systems with a similar parameter set. Variability of the parameters can be related to difficulties related to undescribed local concentration heterogeneities, physiological adaptation of micro-organisms, a microbial population switch, or regulation between multiple AOB pathways. This variability could be due to a dependence of the N2O production pathways on the nitrite (or free nitrous acid-FNA) concentrations and other operational conditions in different systems. This work gives an overview of the potentialities and limits of single AOB pathway models. Indicating in which condition each single pathway model is likely to explain the experimental observations, this work will also facilitate future work on models in which the two main N2O pathways active in AOB are represented together.
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