scholarly journals De novo assembly of the Brown trout (Salmo trutta m. fario) brain and muscle transcriptome: transcript annotation, tissue differential expression profile and SNP discovery

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
J. Fibla ◽  
N. Oromi ◽  
M. Pascual-Pons ◽  
J. L. Royo ◽  
A. Palau ◽  
...  
Keyword(s):  
Differential Expression ◽  
Brown Trout ◽  
Salmo Trutta ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Differential Expression Analysis ◽  
Cdna Libraries ◽  
Significant Differential Expression ◽  
Reference Transcriptome ◽  
Salmonid Species

Abstract Objectives The Brown trout is a salmonid species with a high commercial value in Europe. Life history and spawning behaviour include resident (Salmo trutta m. fario) and migratory (Salmo trutta m. trutta) ecotypes. The main objective is to apply RNA-seq technology in order to obtain a reference transcriptome of two key tissues, brain and muscle, of the riverine trout Salmo trutta m. fario. Having a reference transcriptome of the resident form will complement genomic resources of salmonid species. Data description We generate two cDNA libraries from pooled RNA samples, isolated from muscle and brain tissues of adult individuals of Salmo trutta m. fario, which were sequenced by Illumina technology. Raw reads were subjected to de-novo transcriptome assembly using Trinity, and coding regions were predicted by TransDecoder. A final set of 35,049 non-redundant ORF unigenes were annotated. Tissue differential expression analysis was evaluated by Cuffdiff. A False Discovery Rate (FDR) ≤ 0.01 was considered for significant differential expression, allowing to identify key differentially expressed unigenes. Finally, we have identified SNP variants that will be useful tools for population genomic studies.

scholarly journals Assembly-free rapid differential gene expression analysis in non-model organisms using DNA-protein alignment

2021 ◽  
Author(s):  
Anish M.S. Shrestha ◽  
Joyce Emlyn B. Guiao ◽  
Kyle Christian R. Santiago
Keyword(s):  
Gene Expression ◽  
Differential Expression ◽  
Expression Analysis ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Differential Expression Analysis ◽  
Homology Search ◽  
Model Organisms ◽  
Rna Seq ◽  
Protein Database

AbstractRNA-seq is being increasingly adopted for gene expression studies in a panoply of non-model organisms, with applications spanning the fields of agriculture, aquaculture, ecology, and environment. Conventional differential expression analysis for organisms without reference sequences requires performing computationally expensive and error-prone de-novo transcriptome assembly, followed by homology search against a high-confidence protein database for functional annotation. We propose a shortcut, where we obtain counts for differential expression analysis by directly aligning RNA-seq reads to the protein database. Through experiments on simulated and real data, we show drastic reductions in run-time and memory usage, with no loss in accuracy. A Snakemake implementation of our workflow is available at:https://bitbucket.org/project_samar/samar

scholarly journals Expression profiling of ileal mucosa in asthma reveals upregulation of innate immunity and genes characteristic of Paneth and goblet cells

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Jan K. Nowak ◽  
Marzena Dworacka ◽  
Nazgul Gubaj ◽  
Arystan Dossimov ◽  
Zhumabek Dossimov ◽  
...  
Keyword(s):  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Differential Expression ◽  
Expression Profiles ◽  
Differential Expression Analysis ◽  
Goblet Cells ◽  
Ileal Mucosa ◽  
Significant Differential Expression ◽  
Immune Genes ◽  
Asthma Patients

Abstract Background The expression profiles of the intestinal mucosa have not been comprehensively investigated in asthma. We aimed to explore this in the Correlated Expression and Disease Association Research (CEDAR) patient cohort. Methods Differential expression analysis of ileal, transverse colon, and rectal biopsies were supplemented by a comparison of transcriptomes from platelets and leukocytes subsets, including CD4+, CD8+, CD14+, CD15+, and CD19+ cells. Asthma patients (n = 15) and controls (n = 15) had similar age (p = 0.967), body mass index (p = 0.870), similar numbers of females (80%) and smoking rates (13.3%). Results Significant differential expression was found in the ileum alone, and not in any other cell/tissue types. More genes were found to be overexpressed (1,150) than under-expressed (380). The most overexpressed genes included Fc Fragment of IgG Binding Protein (FCGBP, logFC = 3.01, pFDR = 0.015), Mucin 2 (MUC2, logFC = 2.78, pFDR = 0.015), and Alpha 1B Defensin (DEFA1B, logFC = 2.73, pFDR = 0.024). Gene ontology implicated the immune system, including interleukins 4 and 13, as well as antimicrobial peptides in this overexpression. There was concordance of gene over- (STAT1, XBP1) and underexpression (NELF, RARA) in asthma and Crohn’s disease ileum when our results were compared to another dataset (p = 3.66 × 10–7). Conclusion Ileal mucosa in asthma exhibits a specific transcriptomic profile, which includes the overexpression of innate immune genes, mostly characteristic of Paneth and goblet cells, in addition to other changes that may resemble Crohn’s disease.

scholarly journals RNA-Seq and differential gene expression analysis in Temora stylifera copepod females with contrasting non-feeding nauplii survival rates: an environmental transcriptomics study

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Ennio Russo ◽  
Chiara Lauritano ◽  
Giuliana d’Ippolito ◽  
Angelo Fontana ◽  
Diana Sarno ◽  
...  
Keyword(s):  
Differential Expression ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Survival Rates ◽  
Natural Populations ◽  
Zooplankton Community ◽  
Differentially Expressed ◽  
Diatom Cell ◽  
Functional Interpretation ◽  
Protein Ubiquitination

Abstract Background Copepods are fundamental components of pelagic food webs, but reports on how molecular responses link to reproductive success in natural populations are still scarce. We present a de novo transcriptome assembly and differential expression (DE) analysis in Temora stylifera females collected in the Gulf of Naples, Mediterranean Sea, where this copepod dominates the zooplankton community. High-Throughput RNA-Sequencing and DE analysis were performed from adult females collected on consecutive weeks (May 23rd and 30th 2017), because opposite naupliar survival rates were observed. We aimed at detecting key genes that may have influenced copepod reproductive potential in natural populations and whose expression was potentially affected by phytoplankton-derived oxylipins, lipoxygenase-derived products strongly impacting copepod naupliar survival. Results On the two sampling dates, temperature, salinity, pH and oxygen remained stable, while variations in phytoplankton cell concentration, oxylipin concentration and oxylipin-per-diatom-cell production were observed. T. stylifera naupliar survival was 25% on May 23rd and 93% on May 30th. De novo assembly generated 268,665 transcripts (isoforms) and 120,749 unique ‘Trinity predicted genes’ (unigenes), of which 50% were functionally annotated. Out of the 331 transcript isoforms differentially expressed between the two sampling dates, 119 sequences were functionally annotated (58 up- and 61 down-regulated). Among predicted genes (unigenes), 144 sequences were differentially expressed and 31 (6 up-regulated and 25 down-regulated) were functionally annotated. Most of the significantly down-regulated unigenes and isoforms were A5 Putative Odorant Binding Protein (Obp). Other differentially expressed sequences (isoforms and unigenes) related to developmental metabolic processes, protein ubiquitination, response to stress, oxidation-reduction reactions and hydrolase activities. DE analysis was validated through Real Time-quantitative PCR of 9 unigenes and 3 isoforms. Conclusions Differential expression of sequences involved in signal detection and transduction, cell differentiation and development offered a functional interpretation to the maternally-mediated low naupliar survival rates observed in samples collected on May 23rd. Down-regulation of A5 Obp along with higher quantities of oxylipins-per-litre and oxylipins-per-diatom-cell observed on May 23rd could suggest oxylipin-mediated impairment of naupliar survival in natural populations of T. stylifera. Our results may help identify biomarker genes explaining variations in copepod reproductive responses at a molecular level.

scholarly journals Transcriptional profiling of Impatiens walleriana genes through different stages of downy mildew infection reveals novel genes involved in disease susceptibility

2019 ◽  
Author(s):  
Stephanie Suarez ◽  
Zunaira Afzal Naveed ◽  
Gul Shad Ali
Keyword(s):  
Downy Mildew ◽  
Disease Susceptibility ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Transcriptional Profiling ◽  
Differential Expression Analysis ◽  
Plant Stress ◽  
Differentially Expressed ◽  
Mildew Resistance ◽  
Impatiens Walleriana

AbstractImpatiens downy mildew is a highly destructive disease of Impatiens walleriana, and economically important bedding ornamental crop. This disease is caused by a recently emerged pathogen Plasmopara obducens. Since both the host and pathogen are relatively less studied, there are only a few genomic resources available for both I. walleriana and P. obducens. In this study, we have analyzed transcriptional changes in I. walleriana in response to P. obducens infection during different stages of disease development. Our main goal was to identify candidate genes that may be involved in I. walleriana susceptibility to P. obducens. Since the genome of I. walleriana is not available publicly, we constructed and optimized a de novo transcriptome assembly consisting of 73,022 transcripts. Differential expression analysis based on this optimized de novo transcriptome assembly revealed 3,000 to 4,500 differentially expressed transcripts (DETs) at 0 hr, 12 hr, 48 hr, 120 hr, and 240 hr time points after infection. Functional annotation of these DETs revealed that numerous plant stress responsive genes are activated and deactivated throughout the infection cycle. Genes in the calcium signaling pathways, receptor-like kinases (RLKs) including 10 disease resistance associated RLK transcripts, powdery mildew resistance genes (MLO), and many other plant stress related genes were predominantly differentially expressed in I. walleriana in response to P. obducens. Analyses reported here provides molecular insights into the disease susceptibility mechanism of the Impatiens downy mildew, and lays out a strong foundation for future studies aimed at improving downy mildew resistance in I. walleriana.

scholarly journals Construction of a reference transcriptome for the analysis of male sterility in sugi (Cryptomeria japonica D. Don) focusing on MALE STERILITY 1 (MS1)

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247180
Author(s):  
Fu-Jin Wei ◽  
Saneyoshi Ueno ◽  
Tokuko Ujino-Ihara ◽  
Maki Saito ◽  
Yoshihiko Tsumura ◽  
...  
Keyword(s):  
Male Sterility ◽  
Cryptomeria Japonica ◽  
Evolutionary Biology ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Single Copy ◽  
Reference Transcriptome ◽  
Three Stages ◽  
Significant Expression ◽  
Short Timeframe

Sugi (Cryptomeria japonica D. Don) is an important conifer used for afforestation in Japan. As the genome of this species is 11 Gbps, it is too large to assemble within a short timeframe. Transcriptomics is one approach that can address this deficiency. Here we designed a workflow consisting of three stages to de novo assemble transcriptome using Oases and Trinity. The three transcriptomic stage used were independent assembly, automatic and semi-manual integration, and refinement by filtering out potential contamination. We identified a set of 49,795 cDNA and an equal number of translated proteins. According to the benchmark set by BUSCO, 87.01% of cDNAs identified were complete genes, and 78.47% were complete and single-copy genes. Compared to other full-length cDNA resources collected by Sanger and PacBio sequencers, the extent of the coverage in our dataset was the highest, indicating that these data can be safely used for further studies. When two tissue-specific libraries were compared, there were significant expression differences between male strobili and leaf and bark sets. Moreover, subtle expression difference between male-fertile and sterile libraries were detected. Orthologous genes from other model plants and conifer species were identified. We demonstrated that our transcriptome assembly output (CJ3006NRE) can serve as a reference transcriptome for future functional genomics and evolutionary biology studies.

scholarly journals De novo Transcriptome Assembly of Senna occidentalis Sheds Light on the Anthraquinone Biosynthesis Pathway

2022 ◽  
Vol 12 ◽  
Author(s):  
Sang-Ho Kang ◽  
Woo-Haeng Lee ◽  
Joon-Soo Sim ◽  
Niha Thaku ◽  
Saemin Chang ◽  
...  
Keyword(s):  
De Novo ◽  
Transcriptome Assembly ◽  
Differential Expression Analysis ◽  
Biosynthesis Pathway ◽  
Rna Seq ◽  
Pharmacological Activities ◽  
Secondary Metabolite Biosynthesis ◽  
Long Read ◽  
Gene Expression Levels

Senna occidentalis is an annual leguminous herb that is rich in anthraquinones, which have various pharmacological activities. However, little is known about the genetics of S. occidentalis, particularly its anthraquinone biosynthesis pathway. To broaden our understanding of the key genes and regulatory mechanisms involved in the anthraquinone biosynthesis pathway, we used short RNA sequencing (RNA-Seq) and long-read isoform sequencing (Iso-Seq) to perform a spatial and temporal transcriptomic analysis of S. occidentalis. This generated 121,592 RNA-Seq unigenes and 38,440 Iso-Seq unigenes. Comprehensive functional annotation and classification of these datasets using public databases identified unigene sequences related to major secondary metabolite biosynthesis pathways and critical transcription factor families (bHLH, WRKY, MYB, and bZIP). A tissue-specific differential expression analysis of S. occidentalis and measurement of the amount of anthraquinones revealed that anthraquinone accumulation was related to the gene expression levels in the different tissues. In addition, the amounts and types of anthraquinones produced differ between S. occidentalis and S. tora. In conclusion, these results provide a broader understanding of the anthraquinone metabolic pathway in S. occidentalis.

scholarly journals Investigating Resistance to Emamectin Benzoate in the Tomato Borer Tuta Absoluta

2021 ◽  
Author(s):  
Emmanouil Roditakis ◽  
Marianna Stavrakaki ◽  
Aris Ilias ◽  
Panagiotis Ioannidis ◽  
John Vontas
Keyword(s):  
De Novo ◽  
Transcriptome Assembly ◽  
Differential Expression Analysis ◽  
Tuta Absoluta ◽  
Emamectin Benzoate ◽  
Metabolic Resistance ◽  
Resistance Level ◽  
Specific Chemical ◽  
Potential Implication ◽  
Tomato Borer

Abstract The tomato borer Tuta absoluta is a major pest of tomato mainly controlled by chemical insecticides. However, development of resistance to specific chemical classes has made control of the pest extremely difficult. Emamectin benzoate belongs to the avermectin mode of action and to date, low or no resistance levels against this insecticide have been documented. Recently, reduced efficacy of emamectin benzoate was documented, in a field population from Crete (9-fold resistant ratio (RR)). Subsequent laboratory selections with emamectin benzoate for eight sequential generations, resulted in an increase of the RR to 60-fold, the highest resistance level reported to the particular insecticide. Hereby, we are presenting the characterization of emamectin benzoate resistance in T. absoluta. Sequencing of the GluCl and GABA receptor (rdl) genes, the molecular targets of emamectin benzoate, indicted absence of non-synonymous SNPs. The use of known enzyme inhibitors (PBO, DEF and DEM) revealed that P450s partially synergized emamectin benzoate resistance, suggesting potential implication of metabolic resistance. RNA-seq approach was used to identify differentially expressed genes, from emamectin benzoate resistant and susceptible T. absoluta populations. Twelve libraries were sequenced using the Illumina platform, which generated 81 Gbp, thus substantially increasing the number of publicly available genomic resources for this species. The de novo transcriptome assembly consisted of 549,601 contigs, grouped in 233,453 unigenes. Differential expression analysis and qPCR validation revealed over-expression of one unigene similar to cytochrome P450 (Clan 4) potentially implicated in emamectin benzoate resistance, supporting further the involvement of P450s in the observed resistance phenotype.

Potential of habitat complexity for mitigating interference competition between native and non-native salmonid species

2008 ◽  
Vol 86 (5) ◽  
pp. 386-393 ◽  
Author(s):  
Koh Hasegawa ◽  
Koji Maekawa
Keyword(s):  
Brown Trout ◽  
Habitat Complexity ◽  
Salmo Trutta ◽  
Woody Debris ◽  
Interference Competition ◽  
Salvelinus Leucomaenis ◽  
Salmonid Species ◽  
Local Abundance ◽  
Native White ◽  
White Spotted Char

This study aimed to confirm the mitigation effect of structurally complex habitat on interspecific interference competition between native and non-native salmonid species. We evaluated the effects of habitat complexity (number and size of aggregates of large woody debris and length of undercut banks) and other habitat components that were related to the local abundance of salmonids (dimension and mean depth of pool, mean particle size, and mean current velocity) on the local abundance of native white-spotted char ( Salvelinus leucomaenis (Pallas, 1814)) in allopatry and sympatry with non-native brown trout ( Salmo trutta L., 1758). The field survey was conducted in a Japanese montane stream. The number of char in pool habitats in the allopatric area correlated positively with habitat size, i.e., pool dimension. The number of char in the sympatric area with trout was positively correlated with habitat complexity, while it was negatively correlated with number of trout. In this study, we found that structurally complex habitats might be effective in mitigating interspecific competition between native white-spotted char and non-native brown trout in a montane stream.

scholarly journals RNA-Seq of the Caribbean reef-building coralOrbicella faveolata(Scleractinia-Merulinidae) under bleaching and disease stress expands models of coral innate immunity

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e1616 ◽  
Author(s):  
David A. Anderson ◽  
Marcus E. Walz ◽  
Ernesto Weil ◽  
Peter Tonellato ◽  
Matthew C. Smith
Keyword(s):  
Innate Immunity ◽  
Immune System ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Coral Disease ◽  
Thermal Anomaly ◽  
Molecular Physiology ◽  
Reference Transcriptome ◽  
Caribbean Reef ◽  
The Caribbean

Climate change-driven coral disease outbreaks have led to widespread declines in coral populations. Early work on coral genomics established that corals have a complex innate immune system, and whole-transcriptome gene expression studies have revealed mechanisms by which the coral immune system responds to stress and disease. The present investigation expands bioinformatic data available to study coral molecular physiology through the assembly and annotation of a reference transcriptome of the Caribbean reef-building coral,Orbicella faveolata. Samples were collected during a warm water thermal anomaly, coral bleaching event and Caribbean yellow band disease outbreak in 2010 in Puerto Rico. Multiplex sequencing of RNA on the Illumina GAIIx platform and de novo transcriptome assembly by Trinity produced 70,745,177 raw short-sequence reads and 32,463O. faveolatatranscripts, respectively. The reference transcriptome was annotated with gene ontologies, mapped to KEGG pathways, and a predicted proteome of 20,488 sequences was generated. Protein families and signaling pathways that are essential in the regulation of innate immunity across Phyla were investigated in-depth. Results were used to develop models of evolutionarily conserved Wnt, Notch, Rig-like receptor, Nod-like receptor, and Dicer signaling.O. faveolatais a coral species that has been studied widely under climate-driven stress and disease, and the present investigation provides new data on the genes that putatively regulate its immune system.

scholarly journals RNA-Sequencing based gene expression landscape of guava cv. Allahabad Safeda and comparative analysis to colored cultivars

2020 ◽  
Author(s):  
Amandeep Mittal ◽  
Inderjit Singh Yadav ◽  
Naresh Kumar Arora ◽  
Rajbir Singh Boora ◽  
Meenakshi Mittal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Differential Expression ◽  
Rna Sequencing ◽  
Fruit Development ◽  
De Novo ◽  
Transcriptome Assembly ◽  
White Pulp ◽  
De Novo Transcriptome Assembly ◽  
De Novo Transcriptome ◽  
Tissue Specific

Abstract Background Guava ( Psidium guajava L.) is an important fruit crop of tropical and subtropical areas of the world. Genomics resources in guava are scanty. RNA-Seq based tissue specific expressed genomic information, de novo transcriptome assembly, functional annotation and differential expression among contrasting genotypes has potential to set the stage for the functional genomics for traits of commerce. Results Development of fruit from flower involves orchestration of myriad molecular switches. We did comparative transcriptome sequencing on leaf, flower and fruit tissues of cv. Allahabad Safeda to understand important genes and pathways controlling fruit development. Tissue specific RNA sequencing and de novo transcriptome assembly using Trinity pipeline provided us the first reference transcriptome for guava consisting of 84,206 genes comprising 279,792 total transcripts with N50 of 3,603 bp. Blast2GO assigned annotation to 116,629 transcripts and PFam based HMM profile annotated 140,061 transcripts with protein domains. Differential expression with EdgeR identified 3033 genes in Allahabad Safeda tissues and 68 genes among colored tissue comparisons. Mapping the differentially expressed transcripts over molecular pathways indicate significant hormonal changes during fruit development. Comparisons of red vs green peel in guava cv. Apple Colour, white pulp vs red pulp in Punjab pink and fruit maturation vs ripening in non-coloured Allahabad Safeda indicates up-regulation of ethylene biosynthesis accompanied to secondary metabolism like phenylpropanoid and monolignol pathways. Conclusions Benchmarking Universal Single-Copy Orthologs analysis of de novo transcriptome of guava with eudicots identified 93.7% complete BUSCO genes. In silico differential gene expression among tissue types of Allahabad Safeda and validation of candidate genes with qRT-PCR in contrasting colour genotypes promises the utility of this first guava transcriptome for its potential of tapping the genetic elements from germplasm collections for enhancing fruit traits.

Sign in / Sign up

Export Citation Format

Share Document