Exploring core functional fungi driving the metabolic conversion in the industrial pile fermentation of Qingzhuan tea.

The distinct sensory quality of Qingzhuan tea is mainly formed in pile fermentation by a group of functional microorganisms but the core functional ones was poorly characterized. Therefore, this study investigated the dynamic changes in the fungal community and metabolic profile by integrating microbiomics and metabolomics, and explored the core functional fungi driving the metabolic conversion in the industrial pile fermentation of Qingzhuan tea. Indicated by microbiomics analysis, Aspergillus dominated the entire pile-fermentation process, while Thermoascus, Rasamsonia, and Cylindrium successively abounded in the different stages of the pile fermentation. A total of 50 differentially changed metabolites were identified, with the hydrolysis of galloyl/polymeric catechins, biosynthesis of theabrownins, oxidation of catechins, N-ethyl-2-pyrrolidinone substitution of catechins, and deglycosylation of flavonoid glucosides. Nine fungal genera were identified as core functional fungi, in which Aspergillus linked to the hydrolysis of polymeric catechins and insoluble polysaccharides as well as biosynthesis of theabrownins, while Thermoascus participated in the biosynthesis of theabrownins, deglycosylation of flavonoid glucosides, and N-ethyl-2-pyrrolidinone substitution of catechins. These findings would advance our understanding of the quality formation of Qingzhuan tea and provide a benchmark for precise inoculation for its quality improvement.

Reconstitution of RNA cap methylation reveals different features of SARS-CoV-2 and SARS-CoV methyltransferases.

Cap RNA methylations play important roles in the replication, evasion of host RNA sensor recognition, and pathogenesis. Coronaviruses possess both guanine N7- and 2'-O-ribose methyltransferases (N7-MTase and 2'-O-MTase) encoded by nonstructural protein (nsp) 14 and nsp16/10 complex, respectively. In this study, we reconstituted the two-step RNA methylations of N7-MTase and 2'-O-MTase of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in vitro and demonstrated its common and different features in comparison with that of SARS-CoV. We revealed that the nsp16/10 2'-O-MTase of SARS-CoV-2 has a broader substrate selectivity than the counterpart of SARS-CoV and can accommodate both unmethylated and uncapped RNA substrates in a sequence-independent manner. Most intriguingly, the substrate selectivity of nsp16/10 complex is not determined by the apoenzyme of nsp16 MTase but by its cofactor nsp10. These results provide insight into the unique features of SARS-CoV-2 MTases and may help develop strategies to precisely intervene in the methylation pathway and pathogenesis of SARS-CoV-2.

Monitoring the dynamic changes in aroma during the whole processing of Qingzhuan tea at an industrial scale: From fresh leaves to finished tea.

Aroma is one of the most outstanding quality characteristics of Qingzhuan tea (QZT), but its formation is still unclear. Thus, the volatile organic compounds (VOCs) during the whole processing of QZT were investigated by headspace solid-phase microextraction/gas chromatography-mass spectrometry. Based on 144 identified VOCs, the results showed that de-enzyming, sun-drying, and piling fermentation were the key processes of QZT aroma formation. Furtherly, 42 differential VOCs (VIP > 1.0 and p < 0.05) and 16 key VOCs (rOAV > 1.0 and/or ROAV > 1.0) were screened. Especially, sulcatone and ß-ionone (rOAV > 100 and ROAV > 10) were considered the most important contributors to the aroma of QZT. The metabolisms of key VOCs were mainly involved in oxidative degradation of fatty acids, degradation of carotenoids, and methylation of gallic acid. This study could help to more comprehensively understand the aroma formation in QZT processing at an industrial scale.

Non-Structural Protein 5 of Zika Virus Interacts with p53 in Human Neural Progenitor Cells and Induces p53-Mediated Apoptosis.

Zika virus (ZIKV) infection could disrupt neurogenesis and cause microcephaly in neonates by targeting neural progenitor cells (NPCs). The tumor suppressor p53-mediated cell cycle arrest and apoptotic cell death have been suggested to be activated upon ZIKV infection, yet the detailed mechanism is not well understood. In the present study, we investigated the effects of ZIKV-encoded proteins in the activation of p53 signaling pathway and found that, among the ten viral proteins, the nonstructural protein 5 (NS5) of ZIKV most significantly activated the transcription of p53 target genes. Using the immunoprecipitation-coupled mass spectrometry approach, we identified that ZIKV-NS5 interacted with p53 protein. The NS5-p53 interaction was further confirmed by co-immunoprecipitation and GST pull-down assays. In addition, the MTase domain of NS5 and the C-terminal domain of p53 were mapped to be responsible for the interaction between these two proteins. We further showed that ZIKV-NS5 was colocalized with p53 and increased its protein level in the nuclei and able to prolong the half-life of p53. Furthermore, lentivirus-mediated expression of ZIKV-NS5 in hNPCs led to an apparent cell death phenotype. ZIKV-NS5 promoted the cleavage of PARP1 and significantly increased the cell apoptosis of hNPCs. Taken together, these findings revealed that ZIKV-NS5 is a previously undiscovered regulator of p53-mediated apoptosis in hNPCs, which may contribute to the ZIKV-caused abnormal neurodevelopment.

A Random Regression Model Based on a Single-Step Method for Improving the Genomic Prediction Accuracy of Residual Feed Intake in Pigs.

Residual feed intake (RFI) is considered as a measurement of feed efficiency, which is greatly related to the growth performance in pigs. Daily feeding records can be obtained from automatic feeders. In general, RFI is usually calculated from the total measurement records during the whole test period. This measurement cannot reflect genetic changes in different growth periods during the test. A random regression model (RRM) provides a method to model such type of longitudinal data. To improve the accuracy of genetic prediction for RFI, the RRM and regular animal models were applied in this study, and their prediction performances were compared. Both traditional pedigree-based relationship matrix (A matrix) and pedigree and genomic information-based relationship matrix (H matrix) were applied for these two models. The results showed that, the prediction accuracy of the RRM was higher than that of the animal model, increasing 24.2% with A matrix and 40.9% with H matrix. Furthermore, genomic information constantly improved the accuracy of evaluation under each evaluation model. In conclusion, longitudinal traits such as RFI can describe feed efficiency better, and the RRM with both pedigree and genetic information was superior to the animal model. These results provide a feasible method of genomic prediction using longitudinal data in animal breeding.

The Core Role of Bacillus subtilis and Aspergillus fumigatus in Pile-Fermentation Processing of Qingzhuan Brick Tea.

To identify the microorganisms responsible for the formation of the main quality components of Qingzhuan brick tea (QZBT) during solid-state fermentation (SSF), predominant thermoduric strains were isolated from the tea leaves collected during SSF. According to their capability of releasing cellulase, pectase, protease, and polyphenol oxidase, four strains were selected as starter cultures to ferment sun-dried tea leaves during artificially inoculated SSF. According to the major enzymatic activities and quality components content (tea polyphenols, catechins, amino acids, soluble sugar, and theabrownin), it was found that Aspergillus fumigatus M1 had a significant effect on the transformation of polyphenols and Bacillus subtilis X4 could enhance the ability of bioconversion of strain M1. Strain X4 and M1 may be the core microbes responsible for developing these biochemical components of QZBT, as the values of quality components of tea leaves fermented by these two strains for 6 days was very close to that of the sample naturally fermented for 35 days in the tea factory. The results could be significant in developing industrial starters for the manufacture of QZBT and stabilizing the product quality of different batches.