The Microbiome Protocols eBook initiative: Building a bridge to microbiome research.

The Microbiome Protocols eBook (MPB) serves as a crucial bridge, filling gaps in microbiome protocols for both wet experiments and data analysis. The first edition, launched in 2020, featured 152 meticulously curated protocols, garnering widespread acclaim. We now extend a sincere invitation to researchers to participate in the upcoming 2nd version of MPB, contributing their valuable protocols to advance microbiome research.

Bisphenol A affects microbial interactions and metabolic responses in sludge anaerobic digestion.

The widespread use of bisphenol A (BPA) has resulted in the emergence of new pollutants in various environments, particularly concentrated in sewage sludge. This study investigated the effects of BPA on sludge anaerobic digestion, focusing specifically on the interaction of microbial communities and their metabolic responses. While the influence of BPA on methane accumulation is not significant, BPA still enhanced the conversion of soluble COD, protein, and polysaccharides. BPA also positively influenced the hydrolysis-acidogenesis process, leading to 17% higher concentrations of volatile fatty acids (VFAs). Lower BPA levels (0.2-0.5 mg/kg dw) led to decreased hydrolysis and acidogenesis gene abundance, indicating metabolic inhibition; conversely, higher concentrations (1-5 mg/kg dw) increased gene abundance, signifying metabolic enhancement. Diverse methane metabolism was observed and exhibited alterations under BPA exposure. The presence of BPA impacted both the diversity and composition of microbial populations. Bacteroidetes, Proteobacteria, Firmicutes, and Chloroflexi dominated in BPA-treated groups and varied in abundance among different treatments. Changes of specific genera Sedimentibacter, Fervikobacterium, Blvii28, and Coprothermobacter in response to BPA, affecting hydrolysis and acetogenesis. Archaeal diversity declined while the hydrogenotrophic methanogen Methanospirillum thrived under BPA exposure. BPA exposure enabled microorganisms to form structured community interaction networks and boost their metabolic activities during anaerobic digestion. The study also observed the enrichment of BPA biodegradation pathways at high BPA concentrations, which could interact and overlap to ensure efficient BPA degradation. The study provides insights into the digestion performance and interactions of microbial communities to BPA stress and sheds light on the potential effect of BPA during anaerobic digestion.

Anaerobic selenite-reducing bacteria and their metabolic potentials in Se-rich sediment revealed by the combination of DNA-stable isotope probing, metagenomic binning, and metatranscriptomics.

Microorganisms play a critical role in the biogeochemical cycling of selenium (Se) in aquatic environments, particularly in reducing the toxicity and bioavailability of selenite (Se(IV)). This study aimed to identify putative Se(IV)-reducing bacteria (SeIVRB) and investigate the genetic mechanisms underlying Se(IV) reduction in anoxic Se-rich sediment. Initial microcosm incubation confirmed that Se(IV) reduction was driven by heterotrophic microorganisms. DNA stable-isotope probing (DNA-SIP) analysis identified Pseudomonas, Geobacter, Comamonas, and Anaeromyxobacter as putative SeIVRB. High-quality metagenome-assembled genomes (MAGs) affiliated with these four putative SeIVRB were retrieved. Annotation of functional gene indicated that these MAGs contained putative Se(IV)-reducing genes such as DMSO reductase family, fumarate and sulfite reductases. Metatranscriptomic analysis of active Se(IV)-reducing cultures revealed significantly higher transcriptional levels of genes associated with DMSO reductase (serA/PHGDH), fumarate reductase (sdhCD/frdCD), and sulfite reductase (cysDIH) compared to those in cultures not amended with Se(IV), suggesting that these genes played important roles in Se(IV) reduction. The current study expands our knowledge of the genetic mechanisms involved in less-understood anaerobic Se(IV) bio-reduction. Additinally, the complementary abilities of DNA-SIP, metagenomics, and metatranscriptomics analyses are demonstrated in elucidating the microbial mechanisms of biogeochemical processes in anoxic sediment.

Hydrogel electrodes with conductive and substrate-adhesive layers for noninvasive long-term EEG acquisition.

Noninvasive brain-computer interfaces (BCIs) show great potential in applications including sleep monitoring, fatigue alerts, neurofeedback training, etc. While noninvasive BCIs do not impose any procedural risk to users (as opposed to invasive BCIs), the acquisition of high-quality electroencephalograms (EEGs) in the long term has been challenging due to the limitations of current electrodes. Herein, we developed a semidry double-layer hydrogel electrode that not only records EEG signals at a resolution comparable to that of wet electrodes but is also able to withstand up to 12 h of continuous EEG acquisition. The electrode comprises dual hydrogel layers: a conductive layer that features high conductivity, low skin-contact impedance, and high robustness; and an adhesive layer that can bond to glass or plastic substrates to reduce motion artifacts in wearing conditions. Water retention in the hydrogel is stable, and the measured skin-contact impedance of the hydrogel electrode is comparable to that of wet electrodes (conductive paste) and drastically lower than that of dry electrodes (metal pin). Cytotoxicity and skin irritation tests show that the hydrogel electrode has excellent biocompatibility. Finally, the developed hydrogel electrode was evaluated in both N170 and P300 event-related potential (ERP) tests on human volunteers. The hydrogel electrode captured the expected ERP waveforms in both the N170 and P300 tests, showing similarities in the waveforms generated by wet electrodes. In contrast, dry electrodes fail to detect the triggered potential due to low signal quality. In addition, our hydrogel electrode can acquire EEG for up to 12 h and is ready for recycled use (7-day tests). Altogether, the results suggest that our semidry double-layer hydrogel electrodes are able to detect ERPs in the long term in an easy-to-use fashion, potentially opening up numerous applications in real-life scenarios for noninvasive BCI.

A novel detection technology for early gastric cancer based on Raman spectroscopy.

Despite universal endoscopic screening, early detection of gastric cancer is challenging, led researchers to seek for a novel approach in detecting. Raman spectroscopy measurements as a fingerprint of biochemical structure, enable accurate prediction of gastric lesions non-destructively. This study aimed to evaluate the diagnostic power of Raman spectroscopy in early gastric cancer (EGC), and reveal dynamic biomolecular changes in vitro from normal to EGC. To clarify the biochemical alterations in Correa's cascade, Raman spectra of human normal gastric mucosa, intestinal metaplasia, dysplasia, and adenocarcinoma were compared at tissue and cellular levels based on a self-developed data processing program. For effectively identify EGC, Raman spectroscopy was used combined with multiple machine learning methods, including partial least-squares discriminant analysis (PLS-DA), support vector machine (SVM), and convolutional neural network (CNN) with leave-one-out (LOO) cross validation. A total of 450 Raman spectra were investigated in this study. The upregulation of νsym(O-P-O) backbone (p < 0.001) was identified as a favorable factor for the diagnosis of EGC, the area under the ROC curve (AUC) was up to 0.918. In addition, higher levels of lactic acid (p < 0.001), lipids (p < 0.001), phenylalanine (p = 0.002), and carotenoids (p < 0.001) were detected in EGC. Multivariate machine learning methods for diagnosis of EGC based on Raman spectroscopy, the sensitivity, specificity, accuracy, and AUC were 91.0%, 100%, 94.8%, and 95.8% for SVM, and 84.8%, 92.0%, 88.8%, and 95.5% for CNN, respectively. Raman spectroscopy can be used as a powerful tool for detecting EGC while elucidating biomolecular dynamics in tumorigenesis. (Chictr.org.cn, ChiCTR2200060720.).

Cross-Subject Emotion Recognition Using Fused Entropy Features of EEG.

Emotion recognition based on electroencephalography (EEG) has attracted high interest in fields such as health care, user experience evaluation, and human-computer interaction (HCI), as it plays an important role in human daily life. Although various approaches have been proposed to detect emotion states in previous studies, there is still a need to further study the dynamic changes of EEG in different emotions to detect emotion states accurately. Entropy-based features have been proved to be effective in mining the complexity information in EEG in many areas. However, different entropy features vary in revealing the implicit information of EEG. To improve system reliability, in this paper, we propose a framework for EEG-based cross-subject emotion recognition using fused entropy features and a Bidirectional Long Short-term Memory (BiLSTM) network. Features including approximate entropy (AE), fuzzy entropy (FE), Rényi entropy (RE), differential entropy (DE), and multi-scale entropy (MSE) are first calculated to study dynamic emotional information. Then, we train a BiLSTM classifier with the inputs of entropy features to identify different emotions. Our results show that MSE of EEG is more efficient than other single-entropy features in recognizing emotions. The performance of BiLSTM is further improved with an accuracy of 70.05% using fused entropy features compared with that of single-type feature.

Exogenous Dopamine and MdTyDC Overexpression Enhance Apple Resistance to Fusarium solani.

Fusarium solani, one of the main pathogenic fungi involved in apple replant disease (ARD), is a serious threat to apple growth and development. Dopamine and tyrosine decarboxylase (TyDC), a key enzyme in the dopamine synthesis pathway, have been reported to play an active role in plant responses to biotic and abiotic stresses, but little is known about the functions of dopamine and Malus domestica TyDC (MdTyDC) in the interaction between F. solani and apple roots. In this study, seedlings treated with exogenous dopamine and apple plants overexpressing MdTyDC were inoculated with F. solani; both treatments reduced the root system damage caused by F. solani. After inoculation with F. solani, exogenous dopamine increased dopamine content in the seedlings; alleviated the inhibition of biomass accumulation; increased root metabolic activity, photosynthetic efficiency, and antioxidant enzyme activities; reduced reactive oxygen species accumulation; and upregulated the expression of genes encoding chitinase, ß-1,3-glucanase, and pathogenesis-related proteins. Similar results were observed in MdTyDC-overexpressing apple plants. In addition, the overexpression of MdTyDC increased tyramine content and the deposition of cell wall-bound amines in roots. Overall, our results reveal that exogenous dopamine and overexpression of MdTyDC enhance apple resistance to F. solani, which is an important application for the prevention of ARD.

MdHB-7 Regulates Water Use Efficiency in Transgenic Apple (Malus domestica) Under Long-Term Moderate Water Deficit.

Improved water use efficiency (WUE) promotes plant survival and crop yield under water deficit conditions. Although the plant-specific HD-Zip I transcription factors have important roles in plant adaptation to various abiotic stresses, including water deficit, their functions in regulating WUE of apple (Malus domestica) are poorly understood. We characterized the role of MdHB-7 in WUE regulation by subjecting MdHB-7 transgenic plants to long-term moderate soil water deficit. The long-term WUE (WUEL) of transgenic apple plants with MdHB-7 overexpression or MdHB-7 RNA interference (RNAi) differed significantly from that of control plants. Upregulation of MdHB-7 caused reduced stomatal density, whereas the suppression of MdHB-7 increased stomatal density under both normal and long-term moderate soil water deficit conditions. Moderate reduction in stomatal density helped to improve the WUE of MdHB-7 overexpression transgenic plants, especially under water deficit conditions. MdHB-7 overexpression plants maintained high rates of photosynthesis that were conducive to the accumulation of biomass and the improvement of WUEL. MdHB-7 overexpression also alleviated the inhibition of root growth caused by long-term moderate soil water deficit and improved root vitality and hydraulic conductivity, which were essential for improving plant WUEL. By contrast, MdHB-7 RNA interference reduced the WUEL of transgenic plants by inhibiting these factors under normal and long-term moderate soil water deficit conditions. Taken together, our results provide solid evidence for a crucial role of MdHB-7 in the regulation of apple WUEL and provide new insights for improving the WUE of apple plants under moderate soil water deficit.

The HD-Zip I transcription factor MdHB7-like confers tolerance to salinity in transgenic apple (Malus domestica).

Salinity is a major environmental constraint that substantially limits global agricultural productivity. HD-Zip I transcription factors are involved in plant responses to salt stress, but little is known about the HD-Zip I genes in apple (Malus domestica). Here, we characterized the function of an apple HD-Zip I gene (MdHB7-like) and report that its expression is induced by salt stress. To further explore its role in salt stress, we created MdHB7-like overexpressing and RNAi transgenic apple plants. The overexpression of MdHB7-like improved the photosynthetic performance and reduced ROS and Na+ accumulation under salt stress. Plants that overexpressed MdHB7-like also showed increased accumulation of proline and soluble sugars, which may have played an important role in their salt stress tolerance. RNAi suppression of MdHB7-like had the opposite effects. Together, our results demonstrate that MdHB7-like is an important regulator of salt tolerance in apple. Our results provide new insights for future research on the mechanisms by which MdHB7-like promotes salt tolerance and provide a potential target for molecular breeding in apple.

Genome-wide analysis of the light-harvesting chlorophyll a/b-binding gene family in apple (Malus domestica) and functional characterization of MdLhcb4.3, which confers tolerance to drought and osmotic stress.

In higher plants, the light-harvesting chlorophyll a/b-binding (Lhc) proteins function in multiple processes that are critical to plant growth, development, and abiotic stress response. However, the Lhc gene family has not been well characterized in the important fruit crop, apple (Malus × domestica Borkh.). In this study, we identified 27 Lhc genes in the apple genome. Phylogenetic analysis showed that the Lhc gene family could be classified into three major subfamilies, each of whose members shared similar conserved motifs. Evolutionary analysis indicated that duplicated MdLhc genes were primarily under purifying selection. MdLhcs were expressed at varying levels in all tissues examined and showed different expression patterns under drought stress. The overexpression of MdLhcb4.3 in transgenic Arabidopsis and apple callus enhanced their tolerance to drought and osmotic stress. Taken together, these results demonstrate the important role of Lhc proteins in the regulation of plant resistance to drought and osmotic stress and provide valuable information for further study of Lhc functions in apple.