AMPK Is Involved in Regulating the Utilization of Carbon Sources, Conidiation, Pathogenicity, and Stress Response of the Nematode-Trapping Fungus Arthrobotrys oligospora.

AMP-activated protein kinase (AMPK), a heterotrimeric complex, can sense energy and nutritional status in eukaryotic cells, thereby participating in the regulation of multiple cellular processes. In this study, we characterized the function of the catalytic α-subunit (SNF1) and the two regulatory ß- and γ-subunits (GAL83 and SNF4) of AMPK in a representative nematode-trapping fungus, Arthrobotrys oligospora, by gene knockout, phenotypic analysis, and RNA sequencing. The ability of the AMPK complex mutants (including ΔAosnf1, ΔAogal83, and ΔAosnf4) to utilize a nonfermentable carbon source (glycerol) was reduced, and the spore yields and trap formation were remarkably decreased. Moreover, AMPK plays an important role in regulating stress response and nematode predation efficiency. Transcriptomic profiling between the wild-type strain and ΔAosnf1 showed that differentially expressed genes were enriched for peroxisome, endocytosis, fatty acid degradation, and multilipid metabolism (sphingolipid, ether lipid, glycerolipid, and glycerophospholipid). Meanwhile, a reduced lipid droplet accumulation in ΔAosnf1, ΔAogal83, and ΔAosnf4 mutants was observed, and more vacuoles appeared in the mycelia of the ΔAosnf1 mutant. These results highlight the important regulatory role of AMPK in the utilization of carbon sources and lipid metabolism, as well as providing novel insights into the regulatory mechanisms of the mycelia development, conidiation, and trap formation of nematode-trapping (NT) fungi. IMPORTANCE NT fungi are widely distributed in various ecosystems and are important factors in the control of nematode populations in nature; their trophic mycelia can form unique infectious devices (traps) for capturing nematodes. Arthrobotrys oligospora is a representative NT fungi which can develop complex three-dimensional networks (adhesive networks) for nematode predation. Here, we demonstrated that AMPK plays an important role in the glycerol utilization, conidiation, trap formation, and nematode predation of A. oligospora, which was further confirmed by transcriptomic analysis of the wild-type and mutant strains. In particular, our analysis indicated that AMPK is required for lipid metabolism, which is primarily associated with energy regulation and is essential for trap formation. Therefore, this study extends the functional study of AMPK in NT fungi and helps to elucidate the molecular mechanism of the regulation of trap development, as well as laying the foundation for the development of efficient nematode biocontrol agents.

[Effect of Step Feed on Denitrifying Phosphorus and Nitrate Removal in a Modification of the Two Sludge A2/O-BAF System].

A modification of the two sludge A2/O-BAF system was used to treat low C/N real domestic sewage. In order to improve the utilization of the carbon source, the effects of two step feeds (pre-anoxic zone and anoxic zone) on denitrifying phosphorus and nitrate removal were studied. According to the formula of material balance for COD, the utilization of carbon source was analyzed and evaluated under different ratios of step feed, simultaneously. The results showed that when the ratio of step feed was 7:3 and the influent concentrations of COD, NH4+-N, TN, and TP were 174.99, 58.19, 59.10, and 5.15 mg·L-1, respectively, their effluent concentrations were 29.48, 4.07, 14.10, and 0.40 mg·L-1, and the removal rates were 82.12%, 92.76%, 75.45%, and 91.20%, respectively. It was found that when the ratio of the denitrifying phosphorus accumulation organisms to the phosphorus accumulation organisms(DPAOs/PAOs) was 98.81%, the efficiencies of denitrifying phosphorus and nitrate removal were optimum. By optimizing step feed, the carbon source was utilized effectively, and the efficiencies of nitrogen and phosphorus removal were improved simultaneously. The theoretical basis has thus been provided for the modification of the two sludge A2/O-BAF system to treat low C/N waste water.