TMK4-mediated FIP37 phosphorylation regulates auxin-triggered N6-methyladenosine modification of auxin biosynthetic genes in Arabidopsis.

The dynamics of N6-methyladenosine (m6A) mRNA modification are tightly controlled by the m6A methyltransferase complex and demethylases. Here, we find that auxin treatment alters m6A modification on auxin-responsive genes. Mechanically, TRANSMEMBRANE KINASE 4 (TMK4), a component of the auxin signaling pathway, interacts with and phosphorylates FKBP12-INTERACTING PROTEIN 37 (FIP37), a core component of the m6A methyltransferase complex, in an auxin-dependent manner. Phosphorylation of FIP37 enhances its interaction with RNA, thereby increasing m6A modification on its target genes, such as NITRILASE 1 (NIT1), a gene involved in indole-3-acetic acid (IAA) biosynthesis. 1-Naphthalacetic acid (NAA) treatment accelerates the mRNA decay of NIT1, in a TMK4- and FIP37-dependent manner, which leads to inhibition of auxin biosynthesis. Our findings identify a regulatory mechanism by which auxin modulates m6A modification through the phosphorylation of FIP37, ultimately affecting mRNA stability and auxin biosynthesis in plants.

A Rotating Machinery Fault Diagnosis Method Based on Dynamic Graph Convolution Network and Hard Threshold Denoising.

With the development of precision sensing instruments and data storage devices, the fusion of multi-sensor data in gearbox fault diagnosis has attracted much attention. However, existing methods have difficulty in capturing the local temporal dependencies of multi-sensor monitoring information, and the inescapable noise severely decreases the accuracy of multi-sensor information fusion diagnosis. To address these issues, this paper proposes a fault diagnosis method based on dynamic graph convolutional neural networks and hard threshold denoising. Firstly, considering that the relationships between monitoring data from different sensors change over time, a dynamic graph structure is adopted to model the temporal dependencies of multi-sensor data, and, further, a graph convolutional neural network is constructed to achieve the interaction and feature extraction of temporal information from multi-sensor data. Secondly, to avoid the influence of noise in practical engineering, a hard threshold denoising strategy is designed, and a learnable hard threshold denoising layer is embedded into the graph neural network. Experimental fault datasets from two typical gearbox fault test benches under environmental noise are used to verify the effectiveness of the proposed method in gearbox fault diagnosis. The experimental results show that the proposed DDGCN method achieves an average diagnostic accuracy of up to 99.7% under different levels of environmental noise, demonstrating good noise resistance.

Transcriptome-wide profiling of RNA N4-cytidine acetylation in Arabidopsis thaliana and Oryza sativa.

Acetylation of N4-cytidine (ac4C) has recently been discovered as a novel modification of mRNA. RNA ac4C modification has been shown to be a key regulator of RNA stability, RNA translation, and the thermal stress response. However, its existence in eukaryotic mRNAs is still controversial. In plants, the existence, distribution pattern, and potential function of RNA ac4C modification are largely unknown. Here we report the presence of ac4C in the mRNAs of both Arabidopsis thaliana and rice (Oryza sativa). By comparing two ac4C sequencing methods, we found that RNA immunoprecipitation and sequencing (acRIP-seq), but not ac4C sequencing, was suitable for plant RNA ac4C sequencing. We present transcriptome-wide atlases of RNA ac4C modification in A. thaliana and rice mRNAs obtained by acRIP-seq. Analysis of the distribution of RNA ac4C modifications showed that ac4C is enriched near translation start sites in rice mRNAs and near translation start sites and translation end sites in Arabidopsis mRNAs. The RNA ac4C modification level is positively correlated with RNA half-life and the number of splicing variants. Similar to that in mammals, the translation efficiency of ac4C target genes is significantly higher than that of other genes. Our in vitro translation results confirmed that RNA ac4C modification enhances translation efficiency. We also found that RNA ac4C modification is negatively correlated with RNA structure. These results suggest that ac4C is a conserved mRNA modification in plants that contributes to RNA stability, splicing, translation, and secondary structure formation.

Effects of dietary toxic cyanobacteria and ammonia exposure on immune function of blunt snout bream (Megalabrama amblycephala).

Cyanobacterial blooms caused by water eutrophication have become a worldwide problem. During the degradation of toxic cyanobacterial blooms, elevated ammonia and microcystins concentrations co-occur and exert toxicity on fish. Up to now, the combined effect of microcystins and ammonia on fish immunotoxicity has not been reported. The present study investigated immune responses of blunt snout bream (Megalabrama amblycephala) to dietary toxic cyanobacteria and ammonia exposure. Megalobrama amblycephala were exposed to solutions with different concentrations of NH3-N (0, 0.06, 0.12 mg/L) and fed with diets containing 15% and 30% of toxic cyanobacteria lyophilized powder for 30 d. The microcystins concentration in different organs of Megalobrama amblycephala was in the following sequence: head kidney > liver > intestine > gonad > spleen > gill > trunk kidney > brain > muscle > heart. In both head kidney and spleen, the MC-LR and MC-RR concentration increased significantly with increasing NH3-N concentration. It indicates that NH3-N maybe promote the accumulation of microcystins in immune organs of Megalobrama amblycephala. Meanwhile, broadened peripheral interspace of lymphocytes, nucleus shrivel and edematous mitochondria were observed in head kidney lymphocyte of toxic treatment fish. Moreover, there were significant interactions between dietary toxic cyanobacteria and ammonia exposure on head kidney macrophage phagocytosis activity, respiratory burst activities, total number of white blood cells and the transcriptional levels of sIgM, mIgD and sIgZ genes. Our data clearly demonstrated that dietary toxic cyanobacteria combined with ammonia exposure showed a synergistic effect on Megalobrama amblycephala immunotoxicity.

Glutathione S-transferase in the white shrimp Litopenaeus vannamei: Characterization and regulation under pH stress.

We first expressed a Mu-class GST from white shrimp Litopenaeus vannamei in Escherichia coli, and then characterized the purified recombinant enzyme with respect to the effects of pH, temperature on its catalytic (1-chloro-2, 4-dinitrobenzene-glutathione conjugation) activity. We also analyzed its expression profile in L. vannamei tissues, and assessed changes in Mu-GST expression, GST activity profiles and mortality rates following exposure of white shrimp to low and high pH (5.6 and 9.3, respectively). Realtime-PCR analysis showed that Mu-GST transcripts were expressed in all examined L. vannamei tissues, but were most abundant in the hepatopancreas. At low pH Mu-GST transcript levels in the hepatopancreas were highest after 12 h, and then declined to their original levels after 24 h. After 12 h they were also upregulated in haemocytes, but downregulated in the gills, and unchanged in the stomach following exposure to pH stress. Western blot analyses confirmed that the Mu-GST protein was strongly expressed in the hepatopancreas after 12 h at low pH and remain unchanged in the stomach after exposure to pH stress. pH-Related changes in GST activities in the shrimp hepatopancreas were similar to those displayed by the Mu-GST mRNA and protein profiles. In addition, the mortality of L. vannamei was higher at high pH than at low pH. These results suggest that L. vannamei Mu-GST expression is stimulated by acidic pH and that it may play important roles in detoxification of xenobiotics and antioxidant defenses.