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PROVIDER: PRJNA509511 | ENA |
REPOSITORIES: ENA
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Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to investigate NGS-derived rice leaf transcriptome profiling (RNA-seq) exposed to green rice leafhopper (GRH) and validated RNA-Seq results using quantitative reverse transcription polymerase chain reaction (qRT–PCR). Methods: Rice leaf mRNA profiles of 21-day-old wild-type (WT) and Near Isogenic Line (NIL) rice were generated by deep sequencing, in triplicate, using Illumina’s HiSeq™ 4000 Sequencing System. For assembled genes, longest contigs of the assembled contigs were filtered and clustered into the non-redundant transcripts using CD-HIT-EST (Li et al., 2001; Bolger et al., 2014). qRT–PCR validation was performed using ROX and SYBR Green assays. Results: RNA-Seq results revealed that an average of 1,766,347 (1,240,317 in control and 526,030 GRH treated plants) and 3,676,765 (2,986,748 control and 690,017 GRH) counts per million mapped (CPM) reads were generated from Ilpum and NIL GRH treated plants, respectively. After alignment, more than 75% of all reads were successfully mapped to the reference genome, and 8,859 genes and 15,815,400 transcripts were obtained. GRH infestation caused differential expression of 4265 up-regulated and 4,594 down-regulated genes in Ilpum, while in NIL, 4,479 and 4,380 genes were up- and down-regulated, respectively 48 h post-infestation. A total of 3,424 differentially expressed genes (DEGs, 1,605 up-regulated in Ilpum and down-regulated in NIL; 1,819 genes up-regulated in NIL and down-regulated in Ilpum) were identified. Interestingly, these DEGs were found to be involved in key physiological processes, with interesting molecular functions, including plant defense against diverse biotic stresses, hormone signaling, and other developmental processes. Based on the quantile normalization of the Fragments Per Kilobase of transcript per Million mapped reads (FPKM) values followed by student’s t-test (p <0.05) we identified 3,283 DEGs in Ilpum (1,935 upregulated and 1,348 downregulated) and 2,599 DEGs in NIL (1,621 up-regulated and 978 down-regulated) with at least log2 (logarithm base 2) 2-fold change (Log2FC) in expression upon GRH infestation. Conclusions: The current study has provided the first RNA-Seq dataset and comparative transcriptome profile of GRH responsive genes from Ilpum (GRH-susceptible rice cultivar) and a near isogenic line (NIL) carrying Grh1 gene introduced from Shingwang into Ilpum to investigate the possible transcriptional interaction with other defense related genes. In addition, RNA-Seq data and MapMan analysis revealed the activation of multiple regulatory pathways following GRH infestation. Furthermore, the observed differential transcriptome profile between Ilpum and NIL would suggest the existence of a possible interaction network between Grh1 and other defense related genes towards GRH resistance in rice.
2021-06-10 | GSE176497 | GEO
Project description:We used next-generation sequencing (NGS) to sequence and differentially quantitate miRNAs in 10 pools of plasma derived from individuals with sepsis and SIRS.Plasma pools were preferred to individual samples because they decrease the impact of individual outliers on the analysis. Total RNA was then extracted from equal volumes of plasma and technical duplicates of cDNA libraries for Illumina NGS created. Results from 10 pools representative of 89 individuals (including no-SIRS controls) are uploaded to the repository.
2016-12-01 | E-MTAB-4637 | biostudies-arrayexpress
Project description:Using NGS approach we performed the search of multiple sclerosis-related miRNAs. We used PBMC as an informative and easily accessible biological material. To exclude bias in miRNA expression levels caused by disease modifying therapies, miRNA profiling was performed in treatment-naïve RRMS patients. Taking into account hypothetic gender specificity in disease pathogenesis we compared miRNA expression in RRMS patients and HCs separately for men and women. MiRNA profiling in men identified 32 differentially expressed miRNAs, which passed threshold for multiple corrections and may be attributed to MS-related. At the same time we did not find well-defined MS-specific miRNA expression signatures in women using NGS
2019-01-11 | GSE124900 | GEO
Project description:A loss-of-function mutant in the SKI2 helicase in Arabidopsis was shown to accumulate 5'-cleavage fragments of several miRNA targets. Small RNAs in ski2 single and ski2/rdr6 double mutants (two biological replicates of each genotype) were next analysed by NGS. The data show that several miRNA targets produce RDR6-dependent siRNAs upon inactivation of SKI2. Files containing these NGS data are deposited here. Seeds were surface sterilized and germinated on MS medium for 2 weeks. The seedlings were grown on soil until flowering stage in a 16h light period. The inflorescences were harvested and used to prepare RNA. Small RNA libraries were prepared using the NEBnext Multiplex small RNA library prep kit for Illumina sequencing.
2015-10-20 | E-MTAB-3781 | biostudies-arrayexpress
Project description:CGH was used to map structural variation (introgressed regions) among soybean iron-inefficient NIL IsoClark with its iron-efficient recurrent parent Clark and donor parent T203. Another NIL series HiPro, LoPro was also used in the study.
2011-08-02 | GSE31072 | GEO
Project description:Most proteogenomic approaches for mapping single amino acid polymorphisms (SAPs) require construction of a sample-specific database containing protein variants predicted from the next-generation sequencing (NGS) data. We present a new strategy for direct SAP detection without relying on NGS data. Among the 348 putative SAP peptides identified in an industrial yeast strain, 85.6% of SAP sites were validated by genomic sequencing.
2018-04-18 | PXD003101 | Pride
Project description:Tissue-specific transcriptional profiling of the abscission layer (AL) at the base of young flower in rice using laser micro-dissection: NIL(qSH1) vs. Nipponbare. We used two rice varieties, NIL(qSH1) and Nipponbare. NIL(qSH1) is a nearly isogenic line containing the seed shattering gene qSH1. Seed shattering is easy in NIL(qSH1), but it is not in Nipponbare. So, we used some stages of young flower in NIL(qSH1) and some in Nipponbare. Four regions: 1. abscission layer region of NIL(qSH1), 2. upper abscission region of NIL(qSH1), 3. lower abscission layer region of NIL(qSH1), and 4. abscission layer region of Nipponbare. Sample experiments: NIL(qSH1) AL vs. Nipponbare AL, NIL(qSH1) AL vs. NIL(qSH1) upper region of AL, and NIL(qSH1) AL vs. NIL(qSH1) lower region of AL.
2012-03-31 | E-GEOD-33409 | biostudies-arrayexpress
Project description:In the study, RBSDV accumulation was detected in NIL-S starting from 16 days post inoculation (dpi) and increased linearly thereafter to reach a high level. Concurrently, the viral S10 encoding the coat protein exhibited a dramatic accumulation in NIL-S but not in NIL-R, as did the viral S7-1. We performed RNA-seq analysis at 16 dpi between NIL-S and NIL-R. The differentially expressed genes (DEGs) were identified based on the criteria of fold-change > 2 and P < 0.05. Compared to NIL-R, NIL-S exhibited 24 downregulated and 21 upregulated DEGs. Gene Ontology (GO) term analysis revealed that these DEGs are mainly enriched in one molecular process: oxidoreductase activity, and in three biological processes: biology, oxidation-reduction, and single-organism. KEGG analysis demonstrated significant DEG enrichment in three pathways: photosynthesis-antenna proteins, fatty acid elongation, and diterpenoid biosynthesis.
2024-08-07 | GSE249472 | GEO