scholarly journals Transcriptome profiling of Finnsheep ovaries during out-of-season breeding period

2015 ◽  
Vol 24 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Kisun Pokharel ◽  
Jaana Peippo ◽  
Göran Andersson ◽  
Meng-Hua Li ◽  
Juha Kantanen
Keyword(s):  
Candidate Genes ◽  
Reference Genome ◽  
Transcriptome Assembly ◽  
Transcriptome Profiling ◽  
Breeding Period ◽  
Rna Seq ◽  
Sequence Coverage ◽  
Bioinformatics Pipeline ◽  
Two Samples ◽  
The World

Finnsheep is one of the most prolific sheep breeds in the world. We sequenced RNA-Seq libraries from the ovaries of Finnsheep ewes collected during out of season breeding period at about 30X sequence coverage. A total of 86 966 348 and 105 587 994 reads from two samples were mapped against latest available ovine reference genome (Oarv3.1). The transcriptome assembly revealed 14 870 known ovine genes, including the 15 candidate genes for fertility and out-of-season breeding. In this study we successfully used our bioinformatics pipeline to assemble the first ovarian transcriptome of Finnsheep.

scholarly journals Transcriptome Analysis Reveals Key Genes and Pathways Associated with Mummify Piglets

Genome ◽  
2021 ◽  
Author(s):  
Shujie Wang ◽  
Pingxian Wu ◽  
Kai Wang ◽  
Xiang Ji ◽  
Dong Chen ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Transcriptome Profiling ◽  
Differentially Expressed ◽  
Reproductive Capacity ◽  
Signal Pathways ◽  
Comparative Transcriptome ◽  
Rna Seq ◽  
Promising Candidate ◽  
Pork Consumption ◽  
Differential Genes

China is the country with the largest pork consumption in the world. However, the incidence of high mummify piglets (3-5%) is one of the important factors that cause the slow improvement of pig reproductive capacity, and the genetic mechanism is still unclear. This study aimed to identify candidate genes related to high mummify piglets. RNA-seq technology was used to comparative transcriptome profiling of blood from high piglets mummified and healthy sow at different stages of pregnancy (35d, 56d, 77d and 98d). A total of 137 to 420 DEGs were detected in each stage. Seven differentially expressed genes were significantly differentially expressed at various stages. IL-9R, TLR8, ABLIM3, FSH-α, ASCC1, PRKCZ, and GCK may play an important role in course of mummify piglets. The differential genes we identified between the groups were mainly enriched in immune and inflammation regulation, and others were mainly enriched in reproduction. Considering the function of candidate genes, IL-9R and TLR8 were suggested as the most promising candidate genes involved in mummify piglet traits. We speculate that during pregnancy, it may be the combined effects of the above-mentioned inflammation, immune response, and reproduction-related signal pathways that affect the occurrence of mummifying piglets, and further affect pig reproduction.

scholarly journals The value of genotype-specific reference for transcriptome analyses

2021 ◽  
Author(s):  
Wenbin Guo ◽  
Max Coulter ◽  
Robbie Waugh ◽  
Runxuan Zhang
Keyword(s):  
Alternative Splicing ◽  
Reference Genome ◽  
Transcriptome Assembly ◽  
Specific Reference ◽  
Rna Seq ◽  
High Quality ◽  
Common Reference ◽  
Transcript Quantification ◽  
Gene Level ◽  
Reference Transcript

High quality transcriptome assembly using short reads from RNA-seq data still heavily relies upon reference-based approaches, of which the primary step is to align RNA-seq reads to a single reference genome of haploid sequence. However, it is increasingly apparent that while different genotypes within a species share core genes, they also contain variable numbers of specific genes that are only present a subset of individuals. Using a common reference may thus lead to a loss of genotype-specific information in the assembled transcript dataset and the generation of erroneous, incomplete or misleading transcriptomics analysis results. With the recent development of pan-genome information in many species, it is important that we understand the limitations of single genotype references for transcriptomics analysis. In this study, we quantitively evaluated the advantages of using genotype-specific reference genomes for transcriptome assembly and analysis using cultivated barley as a model. We mapped barley cultivar Barke RNA-seq reads to the Barke genome and to the cultivar Morex genome (common barley genome reference) to construct a genotype specific Reference Transcript Dataset (sRTD) and a common Reference Transcript Datasets (cRTD), respectively. We compared the two RTDs according to their transcript diversity, transcript sequence and structure similarity and the accuracy they provided for transcript quantification and differential expression analysis. Our evaluation shows that the sRTD has a significantly higher diversity of transcripts and alternative splicing events. Despite using a high-quality reference genome for assembly of the cRTD, we miss ca. 40% transcripts present in the sRTD and cRTD only has ca. 70% true assemblies. We found that the sRTD is more accurate for transcript quantification as well as differential expression and differential alternative splicing analysis. However, gene level quantification and comparative expression analysis are less affected by the source RTD, which indicates that analysing transcriptomic data at the gene level may be a reasonable compromise when a high-quality genotype-specific reference is not available.

scholarly journals Siberian sturgeon multi-tissue reference transcriptome database

Database ◽  
2020 ◽  
Vol 2020 ◽  
Author(s):  
Christophe Klopp ◽  
Cédric Cabau ◽  
Gonzalo Greif ◽  
André Lasalle ◽  
Santiago Di Landro ◽  
...  
Keyword(s):  
Reference Genome ◽  
Transcriptome Assembly ◽  
Fish Farming ◽  
Siberian Sturgeon ◽  
Supplementary Information ◽  
Rna Seq ◽  
High Quality ◽  
Reference Transcriptome ◽  
Functional Studies ◽  
Transcriptome Database

Abstract Motivation: Siberian sturgeon is a long lived and late maturing fish farmed for caviar production in 50 countries. Functional genomics enable to find genes of interest for fish farming. In the absence of a reference genome, a reference transcriptome is very useful for sequencing based functional studies. Results: We present here a high-quality transcriptome assembly database built using RNA-seq reads coming from brain, pituitary, gonadal, liver, stomach, kidney, anterior kidney, heart, embryonic and pre-larval tissues. It will facilitate crucial research on topics such as puberty, reproduction, growth, food intake and immunology. This database represents a major contribution to the publicly available sturgeon transcriptome reference datasets. Availability: The database is publicly available at http://siberiansturgeontissuedb.sigenae.org Supplementary information:  Supplementary data are available at Database online.

scholarly journals Scavenger: A pipeline for recovery of unaligned reads utilising similarity with aligned reads

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 1587 ◽  
Author(s):  
Andrian Yang ◽  
Joshua Y. S. Tang ◽  
Michael Troup ◽  
Joshua W. K. Ho
Keyword(s):  
Single Cell ◽  
Genetic Variants ◽  
Reference Genome ◽  
Sequence Similarity ◽  
False Negative ◽  
Rna Seq ◽  
Bioinformatics Pipeline ◽  
Alignment Problem ◽  
Variable Mapping ◽  
Reference Genomes

Read alignment is an important step in RNA-seq analysis as the result of alignment forms the basis for downstream analyses. However, recent studies have shown that published alignment tools have variable mapping sensitivity and do not necessarily align all the reads which should have been aligned, a problem we termed as the false-negative non-alignment problem. Here we present Scavenger, a python-based bioinformatics pipeline for recovering unaligned reads using a novel mechanism in which a putative alignment location is discovered based on sequence similarity between aligned and unaligned reads. We showed that Scavenger could recover unaligned reads in a range of simulated and real RNA-seq datasets, including single-cell RNA-seq data. We found that recovered reads tend to contain more genetic variants with respect to the reference genome compared to previously aligned reads, indicating that divergence between personal and reference genomes plays a role in the false-negative non-alignment problem. Even when the number of recovered reads is relatively small compared to the total number of reads, the addition of these recovered reads can impact downstream analyses, especially in terms of estimating the expression and differential expression of lowly expressed genes, such as pseudogenes.

scholarly journals Slinker: Visualising novel splicing events in RNA-Seq data

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 1255
Author(s):  
Breon Schmidt ◽  
Marek Cmero ◽  
Paul Ekert ◽  
Nadia Davidson ◽  
Alicia Oshlack
Keyword(s):  
Rare Disease ◽  
Human Genome ◽  
Rna Sequencing ◽  
Reference Genome ◽  
Data Driven ◽  
Rna Seq ◽  
Bioinformatics Pipeline ◽  
Link Type ◽  
Muscular Disorders ◽  
Data Driven Approach

Visualisation of the transcriptome relative to a reference genome is fraught with sparsity. This is due to RNA sequencing (RNA-Seq) reads being predominantly mapped to exons that account for just under 3% of the human genome. Recently, we have used exon-only references, superTranscripts, to improve visualisation of aligned RNA-Seq data through the omission of supposedly unexpressed regions such as introns. However, variation within these regions can lead to novel splicing events that may drive a pathogenic phenotype. In these cases, the loss of information in only retaining annotated exons presents significant drawbacks. Here we present Slinker, a bioinformatics pipeline written in Python and Bpipe that uses a data-driven approach to assemble sample-specific superTranscripts. At its core, Slinker uses Stringtie2 to assemble transcripts with any sequence across any gene. This assembly is merged with reference transcripts, converted to a superTranscript, of which rich visualisations are made through Plotly with associated annotation and coverage information. Slinker was validated on five novel splicing events of rare disease samples from a cohort of primary muscular disorders. In addition, Slinker was shown to be effective in visualising deletion events within transcriptomes of tumour samples in the important leukemia gene, IKZF1. Slinker offers a succinct visualisation of RNA-Seq alignments across typically sparse regions and is freely available on Github.

scholarly journals Transcriptomic basis for salt tolerance and disease resistance of silverleaf sunflower revealed by Iso-seq and RNA-seq

2019 ◽  
Author(s):  
Jing Bing ◽  
Yunhe Ling ◽  
Peipei An ◽  
Enshi Xiao ◽  
Chunlian Li ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Salt Tolerance ◽  
Reference Genome ◽  
Transcriptome Assembly ◽  
Future Research ◽  
Rna Seq ◽  
Gene Expressions ◽  
Breeding Programs ◽  
Genomic Variance ◽  
Cultivated Species

Abstract Background Silverleaf sunflower, Helianthus argophyllus , is one of the most important wild species that have been usually used for the improvement of cultivated sunflower. Although a reference genome is now available for the cultivated species, H. annuus , its effect in helping understanding the mechanisms underlying the traits of H. argophyllus is limited by the substantial genomic variance between these two species.Results In this study, we generated a high-quality reference transcriptome of H. argophyllus using Iso-seq strategy. This assembly contains 50,153 unique genes covering more than 91% of the whole genes. Among them, we find 205 genes that are absent in the cultivated species and 475 fusion genes containing components of coding or non-coding sequences from the genome of H. annuus . It is interesting that in line with the strong disease resistance observed for H. argophyllus , these H. argophyllus -specific genes are predominantly related to functions of resistance. We have also profiled the gene expressions in leaf and root under normal or salt stressed conditions and, as a result, find distinct transcriptomic responses to salt stress in leaf and root. Particularly, genes involved in several critical processes including the synthesis and metabolism of glutamate and carbohydrate transport are reversely regulated in leaf and root.Conclusions Overall, this study provided insights into the genomic mechanisms underlying the disease resistance and salt tolerance of silverleaf sunflower and the transcriptome assembly and the genes identified in this study can serve as a complement data resources for future research and breeding programs of sunflowers.

scholarly journals Metabolome and Transcriptome Profiling Reveals Anthocyanin Contents and Anthocyanin-Related Genes of Chimeric Leaves in Ananas comosus Var. Bracteatus

2020 ◽  
Author(s):  
Xuzixin Zhou ◽  
Yanbin Xue ◽  
Meiqin Mao ◽  
Yehua He ◽  
Mark Owusu Adjei ◽  
...  
Keyword(s):  
Transcriptome Profiling ◽  
Ananas Comosus ◽  
Anthocyanin Accumulation ◽  
Rna Seq ◽  
Cut Flower ◽  
Two Samples ◽  
Consistent Change ◽  
Leaf Tissues ◽  
Myb Genes ◽  
Different Parts

Abstract Background: Ananas comosus var. bracteatus is a colorful plant used as a cut flower or landscape ornamental. The unique foliage color of this plant includes both green and red leaves and, as a trait of interest, deserves investigation. In this study, green and red parts of chimeric leaves of Ananas comosus var. bracteatus were sampled and anthocyanin accumulation differences were comparatively analyzed at phenotypic, cellular and molecular levels.Results: A CIELAB results indicated that the a* values and L* values samples had significant differences between two parts. Freehand sections showed that anthocyanin presented limited accumulation in the green leaf tissues but obviously accumulation in the epidermal cells of red tissues. Transcriptomic and metabolomic analyses were performed by RNA-seq and LC-ESI-MS/MS. Among the 508 identified metabolites, 10 kinds of anthocyanins were detected, with 6 significantly different between the two samples. The cyanidin-3,5-O-diglucoside content that accounts for nearly 95.6% in red samples was significantly higher than green samples. RNA-Seq analyses showed that 11 out of 40 anthocyanin-related genes were differentially expressed between the green and red samples. Transcriptome and metabolome correlations were determined by nine quadrant analyses, and 9 anthocyanin-related genes, including MYB5 and MYB82, were correlated with 7 anthocyanin-related metabolites in the third quadrant in which genes and metabolites showing consistent change. Particularly, the PCCs between these two MYB genes and cyanidin-3,5-O-diglucoside were above 0.95. Conclusion: Phenotypic colors are closely related to the tissue structures of different leaf parts of Ananas comosus var. bracteatus, and two MYB transcription factors might contribute to differences of anthocyanin accumulation in two parts of Ananas comosus var. bracteatus chimeric leaves. This study lay a foundation for further researches on functions of MYBs in Ananas comosus var. bracteatus and provides new insights to anthocyanin accumulation in different parts of chimeric leaves.

scholarly journals De Novo Transcriptome Assembly of Isatis indigotica at Reproductive Stages and Identification of Candidate Genes Associated with Flowering Pathways

2018 ◽  
Vol 143 (1) ◽  
pp. 56-66
Author(s):  
Yu Bai ◽  
Ying Zhou ◽  
Xiaoqing Tang ◽  
Yu Wang ◽  
Fangquan Wang ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Molecular Mechanisms ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Orthologous Group ◽  
Rna Seq ◽  
Isatis Indigotica ◽  
Regulatory Pathways ◽  
Kegg Pathways ◽  
Bolting And Flowering

The appropriate timing of bolting and flowering is one of the keys to the reproductive success of Isatis indigotica. Several flowering regulatory pathways have been reported in plant species, but we know little about flowering regulatory in I. indigotica. In the present study, we performed RNA-seq and annotated I. indigotica transcriptome using RNA from five tissues (leaves, roots, flowers, fruit, and stems). Illumina sequencing generated 149,907,857 high-quality clean reads and 124,508 unigenes were assembled from the sequenced reads. Of these unigenes, 88,064 were functionally annotated by BLAST searches against the public protein databases. Functional classification and annotation assigned 55,991 and 23,072 unigenes to 52 gene ontology (GO) terms and 25 clusters of orthologous group (COG) categories, respectively. A total of 19,927 unigenes were assigned to 124 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and 80 candidate genes related to plant circadian rhythm were identified. We also identified a number of differentially expressed genes (DEG) and 91 potential bolting and flowering-related genes from the RNA-seq data. This study is the first to identify bolting and flowering-related genes based on transcriptome sequencing and assembly in I. indigotica. The results provide foundations for the exploration of flowering pathways in I. indigotica and investigations of the molecular mechanisms of bolting and flowering in Brassicaceae plants.

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