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The development of electromagnetic wave absorbers operating in the sub-terahertz (sub-THz) region is necessary in 6G communications. We designed and fabricated a sub-THz metamaterial absorber based on metal microcoils embedded and periodically arranged in a dielectric substrate. The microcoil parameters were optimized by calculating the electromagnetic response of the metamaterial using finite element analysis. An actual metamaterial was then fabricated based on the optimized parameters and characterized using THz time-domain spectroscopy. Our microcoil absorber exhibits an absorptance of >80% and a high shielding performance at about 250â GHz. The resonance frequency can be precisely adjusted by modifying the microcoil array dimensions.
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Typical entomopathogenic filamentous fungi such as Beauveria bassiana infect susceptible hosts via penetration of insect cuticle. The pathogenicity of B. bassiana strain to diverse insect hosts is different. While the molecular mechanisms of B. bassiana adapt to different insects are not well clear. B. bassiana GXsk1011 is a hyper-virulent strain from silkworm, which was investigated on the metabolic responses to three cuticle extracts of Bombyx mori, Helicoverpa armigera and Clanis bilineata at 24 h by RNA-seq method. A total of 638 up- and 400 down-regulated differentially expressed genes (DEGs) were identified in B. bassiana grown on H. armigera compared with B. mori, and 910 up- and 401 down-regulated genes for C. bilineata compared with B. mori. Functional categorization showed that DEGs are mainly involved in metabolic processes, localization, catalytic activity and transporter activity. Analysis of 20 highest fold change genes in DEGs showed that when B. bassiana transferred to non-original hosts as H. armigera and C. bilineata, the adhesion (Mad1), protease (Pr2) and cell surface protein (BBA_09174), etc. were down-regulated. While the class III chitinase ChiA2 (BBA_05353, Bbchi-17), major allergen Asp f 2-like protein (BBA_05395, Bb-f2) and nonribosomal peptide synthase, etc. were up-regulated. The secretory lipase that responded to H. armigera and the phosphate permease responded to C. bilineata were also up-regulated in the Top 20 DEGs. These special expressed genes indicate when the B. bassiana transferred to non-original hosts (or called as non-natural hosts), the strain appeared the changes of metabolic response and infection strategies to adapt to new hosts, and implied the key actions of infected adaptation were to break the barrier of different cuticle chitin component and against the immune stress of hosts. This study provided an insight into the B. bassiana that with wide host ranges how to adapt to infect different insect hosts, which will help us to further understand the pathogenesis of B. bassiana infection.
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Beauveria , Animais , Beauveria/genética , Perfilação da Expressão Gênica , Insetos , Transcriptoma , VirulênciaRESUMO
Endothiapepsin is an aspartic proteinase that was first isolated from the plant pathogenic fungus Endothia parasitica. In previous studies, we reported on three endothiapepsin-like proteins in the entomopathogenic fungus Beauveria bassiana; the genes were up-regulated in B. bassiana hyper-virulent strain GXsk1011 at early stage infection in the silkworm. However, whether these proteins play a role in pathogenicity or not remains unknown. In this study, we cloned one protein, BbepnL-1 gene (BBA-07766), that has 98% homology with B. bassiana strain Bb2860, and expressed it in the yeast Pichia pastoris to investigate its function. The endothiapepsin-like protein is a secreted proteinase of molecular weight approximately 40â¯kDa. It has an N-glycosylation site and a mutation in the C-terminal conserved domain- a Thr was mutated to Gly in B. bassiana GXsk1011 and is different than the endothiapepsin of Endothia parasitica. The recombinant endothiapepsin-like protein showed enzyme activity and degraded the protein components of the silkworm cuticle. To further investigate the activity of the endothiapepsin-like protein, we knocked out the gene BbepnL-1 and showed that the loss of BbepnL-1 reduced the virulence in the silkworm. These results demonstrated that the endothiapepsin-like protein of B. bassiana is a virulence factor.
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Ácido Aspártico Endopeptidases/genética , Beauveria/enzimologia , Bombyx/microbiologia , Proteínas Fúngicas/genética , Interações Hospedeiro-Patógeno , Sequência de Aminoácidos , Animais , Ácido Aspártico Endopeptidases/química , Ácido Aspártico Endopeptidases/metabolismo , Bombyx/crescimento & desenvolvimento , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Larva/crescimento & desenvolvimento , Larva/microbiologia , Alinhamento de SequênciaRESUMO
Genetic variants can influence complex traits by altering gene expression through changes to regulatory elements. However, the genetic variants that affect the activity of regulatory elements in pigs are largely unknown, and the extent to which these variants influence gene expression and contribute to the understanding of complex phenotypes remains unclear. Here, we annotate 90,991 high-quality regulatory elements using acetylation of histone H3 on lysine 27 (H3K27ac) ChIP-seq of 292 pig livers. Combined with genome resequencing and RNA-seq data, we identify 28,425 H3K27ac quantitative trait loci (acQTLs) and 12,250 expression quantitative trait loci (eQTLs). Through the allelic imbalance analysis, we validate two causative acQTL variants in independent datasets. We observe substantial sharing of genetic controls between gene expression and H3K27ac, particularly within promoters. We infer that 46% of H3K27ac exhibit a concomitant rather than causative relationship with gene expression. By integrating GWAS, eQTLs, acQTLs, and transcription factor binding prediction, we further demonstrate their application, through metabolites dulcitol, phosphatidylcholine (PC) (16:0/16:0) and published phenotypes, in identifying likely causal variants and genes, and discovering sub-threshold GWAS loci. We provide insight into the relationship between regulatory elements and gene expression, and the genetic foundation for dissecting the molecular mechanism of phenotypes.
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Histonas , Sequências Reguladoras de Ácido Nucleico , Animais , Suínos/genética , Histonas/genética , Histonas/metabolismo , Fenótipo , Locos de Características Quantitativas , Fígado/metabolismoRESUMO
Regulatory sequences and transposable elements (TEs) account for a large proportion of the genomic sequences of species; however, their roles in gene transcription, especially tissue-specific expression, remain largely unknown. Pigs serve as an excellent animal model for studying genomic sequence biology due to the extensive diversity among their wild and domesticated populations. Here, we conducted an integrated analysis using H3K27ac ChIP-seq, H3K4me3 ChIP-seq, and RNA-seq data from 10 different tissues of seven fetuses and eight closely related adult pigs. We aimed to annotate the regulatory elements and TEs to elucidate their associations with histone modifications and mRNA expression across different tissues and developmental stages. Based on correlation analysis between mRNA expression and H3K27ac and H3K4me3 peak activity, results indicated that H3K27ac exhibited stronger associations with gene expression than H3K4me3. Furthermore, 1.45% of TEs overlapped with either the H3K27ac or H3K4me3 peaks, with the majority displaying tissue-specific activity. Notably, a TE subfamily (LTR4C_SS), containing binding motifs for SIX1 and SIX4, showed specific enrichment in the H3K27ac peaks of the adult and fetal ovaries. RNA-seq analysis also revealed widespread expression of TEs in the exons or promoters of genes, including 4 688 TE-containing transcripts with distinct development stage-specific and tissue-specific expression. Of note, 1 967 TE-containing transcripts were enriched in the testes. We identified a long terminal repeat (LTR), MLT1F1, acting as a testis-specific alternative promoter in SRPK2 (a cell cycle-related protein kinase) in our pig dataset. This element was also conserved in humans and mice, suggesting either an ancient integration of TEs in genes specifically expressed in the testes or parallel evolutionary patterns. Collectively, our findings demonstrate that TEs are deeply embedded in the genome and exhibit important tissue-specific biological functions, particularly in the reproductive organs.
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Elementos de DNA Transponíveis , Transcriptoma , Humanos , Masculino , Camundongos , Animais , Suínos/genética , Elementos de DNA Transponíveis/genética , Histonas/genética , Histonas/metabolismo , RNA Mensageiro , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Transativadores/genética , Transativadores/metabolismoRESUMO
High-quality whole-genome resequencing in large-scale pig populations with pedigree structure and multiple breeds would enable accurate construction of haplotype and robust selection-signature detection. Here, we sequence 740 pigs, combine with 149 of our previously published resequencing data, retrieve 207 resequencing datasets, and form a panel of worldwide distributed wild boars, aboriginal and highly selected pigs with pedigree structures, amounting to 1096 genomes from 43 breeds. Combining with their haplotype-informative reads and pedigree structure, we accurately construct a panel of 1874 haploid genomes with 41,964,356 genetic variants. We further demonstrate its valuable applications in GWAS by identifying five novel loci for intramuscular fat content, and in genomic selection by increasing the accuracy of estimated breeding value by 36.7%. In evolutionary selection, we detect MUC13 gene under a long-term balancing selection, as well as NPR3 gene under positive selection for pig stature. Our study provides abundant genomic variations for robust selection-signature detection and accurate haplotypes for deciphering complex traits in pigs.
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Sus scrofa , Sus scrofa/classificação , Sus scrofa/genética , Animais , Sequenciamento Completo do Genoma , Variação Genética , Estudo de Associação Genômica Ampla , Mucinas/genética , Seleção Genética , Tamanho CorporalRESUMO
Dramatic phenotypic differences between domestic pigs and wild boars (Sus scrofa) provide opportunities to investigate molecular mechanisms underlying the formation of complex traits, including morphology, physiology and behaviour. Most studies comparing domestic pigs and wild boars have focused on variations in DNA sequences and mRNA expression, but not on epigenetic changes. Here, we present a genome-wide comparative study on H3K27ac enhancer activities and the corresponding mRNA profiling in the brain and liver tissues of adult Bama Xiang pigs (BMXs) and Chinese wild boars (CWBs). We identified a total of 1,29,487 potential regulatory elements, among which 11,241 H3K27ac peaks showed differential activity between CWBs and BMXs in at least one tissue. These peaks were overrepresented by binding motifs of FOXA1, JunB, ATF3 and BATF, and overlapped with differentially expressed genes that are involved in female mating behaviour, response to growth factors and hormones, and lipid metabolism. We also identified 4118 nonredundant super-enhancers from ChIP-Seq data on H3K27ac. Notably, we identified differentially active peaks located close to or within candidate genes, including TBX19, MSTN, AHR and P2RY1, which were identified in DNA sequence-based population differentiation studies. This study generates a valuable dataset on H3K27ac profiles of the brain and liver from domestic pigs and wild boars, which helps gain insights into the changes in enhancer activities from wild boars to domestic pigs.
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BACKGROUND: A comprehensive landscape of chromatin states for multiple mammalian tissues is essential for elucidating the molecular mechanism underlying regulatory variants on complex traits. However, the genome-wide chromatin accessibility has been only reported in limited tissue types in pigs. RESULTS: Here we report a genome-wide landscape of chromatin accessibility of 20 tissues in two female pigs at ages of 6 months using ATAC-seq, and identified 557,273 merged peaks, which greatly expanded the pig regulatory element repository. We revealed tissue-specific regulatory elements which were associated with tissue-relevant biological functions. We identified both positive and negative significant correlations between the regulatory elements and gene transcripts, which showed distinct distributions in terms of their strength and distances from corresponding genes. We investigated the presence of transposable elements (TEs) in open chromatin regions across all tissues, these included identifications of porcine endogenous retroviruses (PERVs) exhibiting high accessibility in liver and homology of porcine specific virus sequences to universally accessible transposable elements. Furthermore, we prioritized a potential causal variant for polyunsaturated fatty acid in the muscle. CONCLUSIONS: Our data provides a novel multi-tissues accessible chromatin landscape that serve as an important resource for interpreting regulatory sequences in tissue-specific and conserved biological functions, as well as regulatory variants of loci associated with complex traits in pigs.
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The limited knowledge of genomic noncoding and regulatory regions has restricted our ability to decipher the genetic mechanisms underlying complex traits in pigs. In this study, we characterized the spatiotemporal landscape of putative enhancers and promoters and their target genes by combining H3K27ac-targeted ChIP-Seq and RNA-Seq in fetal (prenatal days 74-75) and adult (postnatal days 132-150) tissues (brain, liver, heart, muscle and small intestine) sampled from Asian aboriginal Bama Xiang and European highly selected Large White pigs of both sexes. We identified 101,290 H3K27ac peaks, marking 18,521 promoters and 82,769 enhancers, including peaks that were active across all tissues and developmental stages (which could indicate safe harbor locus for exogenous gene insertion) and tissue- and developmental stage-specific peaks (which regulate gene pathways matching tissue- and developmental stage-specific physiological functions). We found that H3K27ac and DNA methylation in the promoter region of the XIST gene may be involved in X chromosome inactivation and demonstrated the utility of the present resource for revealing the regulatory patterns of known causal genes and prioritizing candidate causal variants for complex traits in pigs. In addition, we identified an average of 1,124 super-enhancers per sample and found that they were more likely to show tissue-specific activity than ordinary peaks. We have developed a web browser to improve the accessibility of the results ( http://segtp.jxau.edu.cn/pencode/?genome=susScr11 ).
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Sequenciamento de Cromatina por Imunoprecipitação , Genoma , Animais , Feminino , Expressão Gênica , Gravidez , Regiões Promotoras Genéticas/genética , Suínos/genéticaRESUMO
The epigenetic regulation of gene expression is implicated in complex diseases in humans and various phenotypes in other species. There has been little exploration of regulatory elements in the pig. Here, we performed chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-Seq) to profile histone H3 lysine 4 trimethylation (H3K4me3) and histone H3 lysine 27 acetylation (H3K27ac) in the pituitary gland of adult Bama Xiang and Large White pigs, which have divergent evolutionary histories and large phenotypic differences. We identified a total of 65,044 non-redundant regulatory regions, including 23,680 H3K4me3 peaks and 61,791 H3K27ac peaks (12,318 proximal and 49,473 distal), augmenting the catalog of pituitary regulatory elements in pigs. We found 793 H3K4me3 and 3,602 H3K27ac peaks that show differential activity between the two breeds, overlapping with genes involved in the Notch signaling pathway, response to growth hormone (GH), thyroid hormone signaling pathway, and immune system, and enriched for binding motifs of transcription factors (TFs), including JunB, ATF3, FRA1, and BATF. We further identified 2,025 non-redundant super enhancers from H3K27ac ChIP-seq data, among which 302 were shared in all samples of cover genes enriched for biological processes related to pituitary function. This study generated a valuable dataset of H3K4me3 and H3K27ac regions in porcine pituitary glands and revealed H3K4me3 and H3K27ac peaks with differential activity between Bama Xiang and Large White pigs.