GhWER controls fiber initiation and early elongation by regulating ethylene signaling pathway in cotton (Gossypium hirsutum).

Cotton fibers are specialized single-cell trichomes derived from epidermal cells, similar to root hairs and trichomes in Arabidopsis. While the MYB-bHLH-WD40 (MBW) complex has been shown to regulate initiation of both root hairs and trichomes in Arabidopsis, the role of their homologous gene in cotton fiber initiation remains unknown. In this study, we identified a R2R3 MYB transcription factor (TF), GhWER, which exhibited a significant increase in expression within the outer integument of ovule at -1.5 DPA (days post anthesis). Its expression peaked at -1 DPA and then gradually decreased. Knockout of GhWER using CRISPR technology inhibited the initiation and early elongation of fiber initials, resulting in the shorter mature fiber length. Additionally, GhWER interacted with two bHLH TF, GhDEL65 and GhbHLH121, suggesting a potential regulatory complex for fiber development. RNA-seq analysis of the outer integument of the ovule at -1.5 DPA revealed that the signal transduction pathways of ethylene, auxin and gibberellin were affected in the GhWER knockout lines. Further examination demonstrated that GhWER directly activated ethylene signaling genes, including ACS1 and ETR2. These findings highlighted the biological function of GhWER in regulating cotton fiber initiation and early elongation, which has practical significance for improving fiber quality and yield. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-024-01477-6.

A dominant negative mutation of GhMYB25-like alters cotton fiber initiation, reducing lint and fuzz.

Cotton (Gossypium hirsutum) fibers, vital natural textile materials, are single-cell trichomes that differentiate from the ovule epidermis. These fibers are categorized as lint (longer fibers useful for spinning) or fuzz (shorter, less useful fibers). Currently, developing cotton varieties with high lint yield but without fuzz remains challenging due to our limited knowledge of the molecular mechanisms underlying fiber initiation. This study presents the identification and characterization of a naturally occurring dominant negative mutation GhMYB25-like_AthapT, which results in a reduced lint and fuzzless phenotype. The GhMYB25-like_AthapT protein exerts its dominant negative effect by suppressing the activity of GhMYB25-like during lint and fuzz initiation. Intriguingly, the negative effect of GhMYB25-like_AthapT could be alleviated by high expression levels of GhMYB25-like. We also uncovered the role of GhMYB25-like in regulating the expression of key genes such as GhPDF2 (PROTODERMAL FACTOR 2), CYCD3; 1 (CYCLIN D3; 1) and PLD (Phospholipase D), establishing its significance as a pivotal transcription factor in fiber initiation. We identified other genes within this regulatory network, expanding our understanding of the determinants of fiber cell fate. These findings offer valuable insights for cotton breeding and contribute to our fundamental understanding of fiber development.

Toxic effects of combined exposure to cadmium and diclofenac on freshwater crayfish (Procambarus clarkii): Insights from antioxidant enzyme activity, histopathology, and gut microbiome.

In recent years, excessive discharge of pollutants has led to increasing concentrations of cadmium (Cd) and diclofenac (DCF) in water; however, the toxicity mechanism of combined exposure of the two pollutants to aquatic animals has not been fully studied. Procambarus clarkii is an economically important aquatic species that is easily affected by Cd and DCF. This study examined the effects of combined exposure to Cd and DCF on the tissue accumulation, physiology, biochemistry, and gut microflora of P. clarkii. The results showed that Cd and DCF accumulated in tissues in the order of hepatopancreas > gill > intestine > muscle. The hepatopancreas and intestines were subjected to severe oxidative stress, with significantly increased antioxidant enzyme activity. Pathological examination revealed lumen expansion and epithelial vacuolisation in the hepatopancreas and damage to the villous capillaries and wall in the intestine. The co-exposure to Cadmium (Cd) and Diclofenac (DCF) disrupts the Firmicutes/Bacteroidetes (F/B) ratio, impairing the regular functioning of intestinal microbiota in carbon (C) and nitrogen (N) cycling. This disturbance consequently hinders the absorption and utilization of energy and nutrients in Procambarus clarkii. This study offers critical insights into the toxicological mechanisms underlying the combined effects of Cd and DCF, and suggests potential approaches to alleviate their adverse impacts on aquatic ecosystems.

Periodontal ligament cells-derived exosomes promote osteoclast differentiation via modulating macrophage polarization.

Several studies have demonstrated that exosomes (Exos) are involved in the regulation of macrophage polarization and osteoclast differentiation. However, the characteristics as well as roles of exosomes from human periodontal ligament cells (hPDLCs-Exos) in M1/M2 macrophage polarization and osteoclast differentiation remain unclear. Here, periodontal ligament cells were successfully extracted by method of improved Type-I collagen enzyme digestion. hPDLCs-Exos were extracted by ultracentrifugation. hPDLCs-Exos were identified by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and western blotting (WB). Osteoclast differentiation was evaluated by real-time quantitative polymerase chain reaction (RT-qPCR), WB and tartrate-resistant acid phosphatase (TRAP) staining. M1/M2 macrophage polarization were evaluated by RT-qPCR and WB. The results showed hPDLCs-Exos promoted osteoclast differentiation and M2 macrophage polarization, but inhibited M1 macrophage polarization. Moreover, M1 macrophages inhibited osteoclast differentiation, whereas M2 macrophages promoted osteoclast differentiation. It has shown that hPDLCs-Exos promoted osteoclast differentiation by inhibiting M1 and promoting M2 macrophage polarization.

Regulatory controls of duplicated gene expression during fiber development in allotetraploid cotton.

Polyploidy complicates transcriptional regulation and increases phenotypic diversity in organisms. The dynamics of genetic regulation of gene expression between coresident subgenomes in polyploids remains to be understood. Here we document the genetic regulation of fiber development in allotetraploid cotton Gossypium hirsutum by sequencing 376 genomes and 2,215 time-series transcriptomes. We characterize 1,258 genes comprising 36 genetic modules that control staged fiber development and uncover genetic components governing their partitioned expression relative to subgenomic duplicated genes (homoeologs). Only about 30% of fiber quality-related homoeologs show phenotypically favorable allele aggregation in cultivars, highlighting the potential for subgenome additivity in fiber improvement. We envision a genome-enabled breeding strategy, with particular attention to 48 favorable alleles related to fiber phenotypes that have been subjected to purifying selection during domestication. Our work delineates the dynamics of gene regulation during fiber development and highlights the potential of subgenomic coordination underpinning phenotypes in polyploid plants.

The early function of cortisol in liver during Aeromonas hydrophila infection: Dynamics of the transcriptome and accessible chromatin landscapes.

In China, channel catfish (Ictalurus punctatus) is an important aquaculture species; however, haemorrhagic disease (Aeromonas hydrophila induced disease) in these fish has caused tremendous economic loss due to high morbidity and mass mortality in the breeding industry. The role of cortisol in bacterial diseases, particularly in the acute phase, remains unclear. In this study, liver transcriptome (RNA-seq) and chromatin accessibility (ATAC-seq) analyses were employed to investigate the early functional role of cortisol in Aeromonas hydrophila-stimulated responses. Our experiments confirmed that A. hydrophila infection can initially significantly increase serum cortisol levels at 1 h after infection. At this time point, the increased serum cortisol levels can significantly regulate A. hydrophila-regulated genes by affecting both transcriptome and chromatin accessibility. Cross-analysis of RNA-seq and ATAC-seq revealed that a certain gene group (92 target_DEGs) was regulated at an early time point by cortisol. KEGG enrichment analysis revealed that the top three pathways according to target_DEGs were cancer, glutathione metabolism, and the Notch signalling pathway. The protein-protein interaction analysis of target_DEGs revealed that they may be primarily involved in cell proliferation, CD8+ T cell function, glutathione synthesis, and activation of the NF-κB signalling pathway. This suggests that after the emergence of immune stress, the early regulation of cortisol is positive against the immune response. It is possible that in this situation, the animal is attempting to avoid dangerous situations and risks and then cope with the imbalance produced by the stressor to ultimately restore homeostasis. Our results will contribute to future research on fish and provide valuable insight regarding the mechanism of immune regulation by cortisol and the study of bacterial haemorrhagic disease in channel catfish.

A positive feedback loop between two C-type lectins originated from gene duplication and relish promotes the expression of antimicrobial peptides in Procambarus clarkii.

Gene duplication (GD) leads to the expansion of gene families that contributes organisms adapting to stress or environment and dealing with the infection of various pathogens. C-type lectins (CTLs) in crustaceans undergo gene expansion and participate in various immune responses. However, the functions of different CTL produced by GD are not fully characterized. In the present study, two CTL genes (designated as PcLec-EPS and PcLec-QPS, respectively) were identified from Procambarus clarkii. PcLec-EPS and PcLec-QPS originate from GD and the main difference between them is exon 3. PcLec-EPS and PcLec-QPS respectively contains EPS and QPS motif in their carbohydrate recognition domain. The mRNA levels of PcLec-EPS and PcLec-QPS in hemocytes, gills, intestine and lymph underwent time-dependent enhancement after D-Mannose and D-Galactose challenge. Recombinant PcLec-EPS and PcLec-QPS could bind to carbohydrates and microbes, and agglutinate bacteria. The results of experiments on recombinant protein injection and RNA interference indicate that PcLec-EPS and PcLec-QPS can respectively strong recognize and bind D-Mannose and D-Galactose, activate the Relish transcriptional factor, and further upregulate the expression of different antimicrobial peptides (AMPs). In addition, these two CTLs and Relish could positively regulate the expression of each other, suggesting that there is a positive feedback loop between two CTLs and Relish that regulates the expression of AMPs. It may contribute to the expansion of the immune response for host quickly and efficiently eliminating pathogenic microorganisms. This study provides new knowledge for clear understanding the significance and function of different CTL generated by GD in immune defenses in crustacean.

Single-cell RNA-seq reveals fate determination control of an individual fibre cell initiation in cotton (Gossypium hirsutum).

Cotton fibre is a unicellular seed trichome, and lint fibre initials per seed as a factor determines fibre yield. However, the mechanisms controlling fibre initiation from ovule epidermis are not understood well enough. Here, with single-cell RNA sequencing (scRNA-seq), a total of 14 535 cells were identified from cotton ovule outer integument of Xu142_LF line at four developmental stages (1.5, 1, 0.5 days before anthesis and the day of anthesis). Three major cell types, fibre, non-fibre epidermis and outer pigment layer were identified and then verified by RNA in situ hybridization. A comparative analysis on scRNA-seq data between Xu142 and its fibreless mutant Xu142 fl further confirmed fibre cluster definition. The developmental trajectory of fibre cell was reconstructed, and fibre cell was identified differentiated at 1 day before anthesis. Gene regulatory networks at four stages revealed the spatiotemporal pattern of core transcription factors, and MYB25-like and HOX3 were demonstrated played key roles as commanders in fibre differentiation and tip-biased diffuse growth respectively. A model for early development of a single fibre cell was proposed here, which sheds light on further deciphering mechanism of plant trichome and the improvement of cotton fibre yield.

The effect of a polystyrene nanoplastic on the intestinal microbes and oxidative stress defense of the freshwater crayfish, Procambarus clarkii.

The widespread generation and accumulation of plastic waste has become a globally recognized problem. However, there are limited reports on the adverse effects of nanomaterials on freshwater crustaceans. This study tested the acute effects of different concentrations (0, 5, 10, and 20 mg/L) after 48 h exposure of 75 nm polystyrene nanoplastic on intestinal microbes, and oxidative stress parameters of freshwater crayfish, Procambarus clarkii. High-throughput sequencing analysis revealed the richness, diversity, and composition of intestinal microbiota in P. clarkii exposed to polystyrene nanoplastic. At the genus level, abundances of Lactobacillus, Faecalibaculum, Niveibacterium, and Candidatus Bacilloplasma were significantly different. The reduced abundance of Lactobacillus could affect the balance of intestinal microbes through quantitative disadvantage, which may lead to reduced immunity of P. clarkii. Streptococcus salivarius, Clostridium butyricum and Lachnospiraceae bacterium10-1 in intestinal tract reached maximum abundance at a polystyrene concentration of 20 mg/L. The increase in the number of some pathogenic bacteria may upset the balance of intestinal microorganisms through the number of dominance, and the decrease in the relative abundance of lactic acid bacteria. Probiotics, such as Lactobacillus salivarius, Lactobacillus murinus, Lactobacillus gasseri, Lactobacillus reuteri, Lactobacillus iners AB-1, and Lactobacillus crispatus in the intestinal tract reached the lowest value at a concentration of 10 mg/L. The reduced abundance of Lactobacillus can affect the balance of intestinal microbes through quantitative disadvantage, which may lead to reduced immunity in P. clarkii. At nanoplastic 10 mg/L, the relative abundance of intestinal pathogens increased, while the relative abundance of lactic acid bacteria and other probiotics decreased. With increases in nanoplastic concentrations, the values of glutathione (GSH), superoxide dismutase (SOD), acid phosphatase (ACP), lysozyme (LZM), alkaline phosphatase (AKP), peroxidase (POD), glutathione peroxidase (GPX), and protein carbonylation were significantly changed. Our data suggested that Lactobacillus may play an adjunctive role in the treatment of oxidative stress in P. clarkii exposed to 75 nm polystyrene. This study represents an important step towards a better understanding of the toxic effects of nanoplastics on aquatic crustaceans.

Full-Length Transcriptome of Red Swamp Crayfish Hepatopancreas Reveals Candidate Genes in Hif-1 and Antioxidant Pathways in Response to Hypoxia-Reoxygenation.

Red swamp crayfish is particularly prone to exposure to hypoxia-reoxygenation stress on account of the respiration and rhythmic, light-dependent photosynthetic activity of the algae and aquatic grass. Up to now, the regulation mechanisms of the adverse effects of hypoxia-reoxygenation for this species were still unknown, especially the roles of the antioxidant enzymes in reducing oxidative damage during reoxygenation. To screen for vital genes or pathways upon hypoxic-reoxygenation stress, hepatopancreas gene expression profiles were investigated by using a strategy combining second and third generation sequencing. Five groups of samples, including hypoxia for 1 and 6 h with DO of 1.0 mg/L, reoxygenation for 1 and 12 h with DO of 6.8 mg/L, and the samples under normoxia condition, were used for transcriptome sequencing. Twenty Illumina cDNA libraries were prepared to screen for the differentially expressed genes (DEGs) among the 5 groups of samples. Based on the assembled reference full-length transcriptome, 389 and 533 significantly DEGs were identified in the groups under severe hypoxia treatment for 1 and 6 h, respectively. The top three enriched pathways for these DEGs were "protein processing in endoplasmic reticulum," "MAPK signaling pathway," and "endocytosis." Among these DEGs, hypoxia-inducible factor 1α (Hif-1α) and some Hif-1 downstream genes, such as Ugt-1, Egfr, Igfbp-1, Pk, and Hsp70, were significant differentially expressed when exposed to hypoxia stress. A series of antioxidant enzymes, including two types of superoxide dismutase (Cu/ZnSOD and MnSOD), catalase (CAT), and glutathione peroxidase (GPx), were identified to be differentially expressed during hypoxia-reoxygenation treatment, implying their distinct modulation roles on reoxygenation-induced oxidative stress. The full-length transcriptome and the critical genes characterized should contribute to the revelation of intrinsic molecular mechanism being associated with hypoxia/reoxygenation regulation and provide useful foundation for future genetic breeding of the red swamp crayfish.

[Research status of Epstein Barr virus in children with adenotonsillar hypertrophy].

Airway obstruction caused by adenotonsillar hypertrophy is one of the most common otolaryngological diseases in children. In recent years, Epstein Barr virus has been found to be closely related to adenotonsillar hypertrophy. This review summarizes the mechanism and epidemiology of adenotonsillar hypertrophy and obstructive sleep apnea syndrome caused by Epstein Barr virus.

Cotton pan-genome retrieves the lost sequences and genes during domestication and selection.

BACKGROUND: Millennia of directional human selection has reshaped the genomic architecture of cultivated cotton relative to wild counterparts, but we have limited understanding of the selective retention and fractionation of genomic components. RESULTS: We construct a comprehensive genomic variome based on 1961 cottons and identify 456 Mb and 357 Mb of sequence with domestication and improvement selection signals and 162 loci, 84 of which are novel, including 47 loci associated with 16 agronomic traits. Using pan-genome analyses, we identify 32,569 and 8851 non-reference genes lost from Gossypium hirsutum and Gossypium barbadense reference genomes respectively, of which 38.2% (39,278) and 14.2% (11,359) of genes exhibit presence/absence variation (PAV). We document the landscape of PAV selection accompanied by asymmetric gene gain and loss and identify 124 PAVs linked to favorable fiber quality and yield loci. CONCLUSIONS: This variation repertoire points to genomic divergence during cotton domestication and improvement, which informs the characterization of favorable gene alleles for improved breeding practice using a pan-genome-based approach.

The differences in plasma/serum ghrelin levels between obstructive sleep apnea-hypopnea patients and controls: A protocol for systematic review and meta-analysis.

BACKGROUND: The association between obstructive sleep apnea-hypopnea syndrome (OSAHS) and plasma/serum ghrelin levels remains controversial. We performed a meta-analysis to evaluate the difference in plasma/serum ghrelin levels between OSAHS patients and controls. METHODS: Database of PubMed, SCI, and Elsevier were searched entirely. Two independents identified eligible studies of ghrelin levels in OSAHS patients. ReviewManager (version 5.3) was adopted for data synthesis. RESULTS: The meta-analysis A pooled the comparison of ghrelin concentrations in OSAHS patients and controls, which included 7 studies and involving 446 participants. The result of the meta-analysis A indicated that plasma/serum ghrelin levels were no significant differences between the OSAHS group and the control group (standard mean difference (SMD) = 0.08, 95% confidence interval (CI) = -0.12 to 0.28, P = .43). As a supplementary, meta-analysis B pooled the comparison of plasma/serum ghrelin levels in OSAHS patients before and after continuous positive airway pressure (CPAP) therapy, which included 155 participants from 4 studies, it revealed that plasma/serum ghrelin levels were no significant differences between before and after CPAP therapy (SMD = 0.12, 95%CI = -0.07 to 0.31, P = .22). CONCLUSION: The meta-analysis A demonstrated that plasma/serum ghrelin levels were no significant differences between the OSAHS group and the control group. The meta-analysis B showed plasma/serum ghrelin levels have no significant changes after CPAP therapy in OSAHS patients.

Genomic organization of the molt-inhibiting hormone gene in the red swamp crayfish Procambarus clarkii and characterization of single-nucleotide polymorphisms associated with growth.

Molt-inhibiting hormone (MIH), a neuropeptide synthesized in the eyestalk in crustaceans, is mainly responsible for the molting by negatively controlling the ecdysteroids secretion. Although there are several reports of the isolation and protein sequencing of MIH in the red swamp crayfish, little is known about the nucleotide sequence and gene organization of this neuropeptide, even less about the association of MIH polymorphisms and growth traits. Here, a 1237 bp full-length MIH cDNA was obtained from the crayfish eyestalk, which encodes a putative protein of 106 amino acids, with a 191 bp 5'-UTR and a 728 bp 3'-UTR. The MIH genomic DNA sequence is 4205 bp in length, which includes three exons interrupted by two introns, and a 929 bp 5'-flanking region. Potential transcription initiation site and transcription factor binding sites were identified in the 5'-flanking region, implying a potential role in transcriptional regulation. Seventeen SNPs in the 5'-flanking region and 3'-UTR were identified, and the associations between these SNPs and growth traits were evaluated with a two-stage design. A SNPs g. -12C > G that showed a significant association with body weight was identified. Individuals with GG genotype had a significantly higher body weight than those with CC genotype (43.98 ±â€¯9.82 g vs. 34.27 ±â€¯6.87 g; P ﹤ 0.001), indicating a beneficial effect of the G allele on the growth of red swamp crayfish. The obtained MIH gene, as well as the identified SNPs, may serve as targets for molecular marker-aided selection in growth improvement of the red swamp crayfish in future studies.

Genome-wide identification, phylogeny and expressional profile of the Sox gene family in channel catfish (Ictalurus punctatus).

The Sox gene family has been systematically characterized in some fish species but not in catfish Ictalurus punctatus. In this study, 25 Sox genes were identified in the channel catfish genome and classified into seven families based on their conserved domains as follows: eight genes in SoxB group (six in SoxB1 subgroup and two in SoxB2 subgroup); five genes in SoxC group; three genes in SoxD and SoxF groups; four genes in SoxE group; and one gene in SoxH and SoxK groups. The mammalian Sox groups SoxA, G, I, and J were not present in catfish. The number of introns in channel catfish Sox genes varied from zero to 13. Sox genes were distributed unevenly across 17 chromosomes. Five members of the ancestral vertebrate Sox genes (Sox1, Sox4, Sox9, Sox11 and Sox19) experienced teleost-specific whole genome duplication during evolution, and now have two copies on different chromosomes. Expression profiles analyses indicated that the accumulation of Sox genes was associated with different tissues, and the expression pattern also differed among each Sox gene group and duplicated gene. This study constitutes a comprehensive overview of the Sox gene family in channel catfish and provides new insights into the evolution of this gene family.

A new microsporidium, Potaspora macrobrachium n.sp. infecting the musculature of pond-reared oriental river prawn Macrobrachium nipponense (Decapoda: Palaemonidae).

This paper described a novel microsporidian infection in the pond-reared oriental river prawn Macrobrachium nipponense. A conspicuous symptom of the infection was progressive white opacity associated with the musculature. Although neither bacteria nor viruses were detected in routine diagnostic tests, apparently degenerated microsporidian cells or spores were frequently observed in wet smears of the musculature from diseased prawns. Histological observations also revealed characteristics typical of microsporidian infection throughout the host. Transmission electron microscopy revealed multiple life stages of a microsporidian parasite within the cytoplasm of host muscle cells. In addition, partial small subunit ribosomal RNA (SSU rRNA) gene was obtained by a nested PCR using microsporidian specific primers. A consensus sequence was then deposited in GenBank (accession no. KU307278) and subjected to a general BLASTn search that yielded hits only for microsporidian sequence records. Phylogenetic analysis showed that the isolate was most similar to the fish microsporidian clade containing the genera Kabatana, Microgemma, Potaspora, Spraguea, and Teramicra. The highest sequence identity, 87%, was with Potaspora spp. Based on histological, ultrastructure and molecular phylogenetic data, we erected a new species, Potaspora macrobrachium for the novel microsporidium. The description of microsporidium in this important commercial host was fundamental for future consideration of factors affecting stock health and sustainability.

The first detection of white spot syndrome virus in naturally infected cultured Chinese mitten crabs, Eriocheir sinensis in China.

An epidemic with a high mortality rate (80-100%) recently occurred in the cultured Chinese mitten crab, Eriocheir sinensis, which is a very important economic crustacean species in China. Using negative stain, histopathology and nested PCR supplemented by sequencing we identified white spot syndrome virus (WSSV) in these crabs. Challenge experiments revealed that the disease was caused by WSSV and confirmed the crab's susceptibility to this virus, which was consistent with previous laboratory-based studies. A cumulative mortality of 100% was observed within 10 days post WSSV injection. This is the first report of WSSV-associated disease outbreaks in the Chinese mitten crab, which is normally reported as an important penaeid-shrimp viral pathogen. Furthermore, this is only the second report to describe a significant pathogen in pond-cultured E. sinensis. These results will enhance the early diagnosis of WSSV in the crab farms and help in monitoring efforts directed at determining the prevalence of the virus in E. sinensis.