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Unknown,Transcriptomics,Genomics,Proteomics

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MRNA-Seq for homoeologous transcript quantitation in polyploid Brassica napus

ABSTRACT: Illumina mRNA-Seq is comparable to microarray analysis for transcript quantification but has increased sensitivity and, importantly, the potential to distinguish between homoeologous genes in polyploids. Using a novel curing process, we adapted a reference sequence that was a consensus derived from ESTs from both Brassica A and C genomes to one containing A and C genome versions for each of the 94,558 original unigenes. We aligned reads from Brassica napus to this cured reference, finding 38% more reads mapping in resynthesised lines and 28% in natural lines. Where the A and C versions differed at single nucleotide positions, termed inter-homoeologue polymorphisms (IHPs), we were able to apportion expression in the polyploid to the A or C genome homoeologues. 43,761 unigenes contained at least one IHP, with a mean frequency of 10.5 per kb unigene sequence. 6,350 of the unigenes with IHPs were differentially expressed between homoeologous gene pairs in resynthesised B. napus. 3,212 unigenes showed a similar pattern of differential expression across a range of natural B. napus crop varieties and, of these, 995 were in common with resynthesised B. napus. Functional classification showed over-representation in gene ontology categories not associated with dosage-sensitivity.

INSTRUMENT(S): Illumina Genome Analyzer IIx

ORGANISM(S): Brassica rapa

SUBMITTER: Robert Davey

PROVIDER: E-MTAB-4340 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Associative transcriptomics of traits in the polyploid crop species Brassica napus.

Harper Andrea L AL Trick Martin M Higgins Janet J Fraser Fiona F Clissold Leah L Wells Rachel R Hattori Chie C Werner Peter P Bancroft Ian I

Nature biotechnology 20120801 8

PMID: 22820317

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Project description:Homoeologous recombination in Brassica napus

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Subgenome-dominant expression and alternative splicing in response to pathogen stress in polyploid Brassica napus and progenitors

Project description:Polyploidy has played an extensive role in the evolution of flowering plants. Allopolyploids, with subgenomes containing duplicated gene pairs called homeologs, can show rapid transcriptome changes including novel alternative splicing (AS) patterns. The extent to which abiotic stress modulates AS of homeologs is a nascent topic in polyploidy research. We subjected both natural and resynthesized lines of polyploid Brassica napus, along with the progenitors B. rapa and B. oleracea, to infection with the fungal pathogen Sclerotinia sclerotiorum. RNA-seq analyses revealed widespread divergence between polyploid subgenomes in both gene expression and AS patterns. Resynthesized B. napus displayed significantly more A and C subgenome biased homeologs under pathogen infection than during uninfected growth. Differential AS (DAS) in response to infection was highest in natural B. napus (12,709 DAS events) and lower in resynthesized Brassica napus (8,863 DAS events). Natural B. napus had more up-regulated events and fewer down-regulated events. There was a global expression bias towards the B. oleracea-derived (C) subgenome in both resynthesized and natural B. napus, enhanced by widespread non-parental downregulation of the B. rapa-derived (A) homeolog. In the resynthesized B. napus specifically, this resulted a disproportionate C subgenome contribution to pathogen defense response, characterized by biases in both transcript expression levels and the proportion of induced genes. Our results elucidate the complex ways in which Sclerotinia infection affects expression and AS of homeologous genes in natural and resynthesized B. napus, and indicate that abiotic stress can influence the evolution of homeologous genes in polyploids.

2023-02-04 | GSE201225 | GEO

Transcription profiling of Brassica lines

Project description:Transcription profiling of Brassica rapa, Brassica oleracea and Brassica napus I and II The nuclear genomes of the resynthesised B. napus lines should be identical but, as one (B. napus I) involved a cross of B. oleracea onto B. rapa, and the other (B. napus II) involved a cross of B rapa onto B. oleracea, they differ in cytoplasm, and hence contain different chloroplast and mitochondrial genomes.

2009-05-03 | GSE15915 | GEO

Gene expression profilling of six species of Brassica and early four generation of resynthesized B. napus (F1-F4)

Project description:Analysis of the different gene expression profiles of natural and resynthesized Brassica polyploids with Illumina deep sequencing technology could help to improve our knowledge of polyploid genome evolution. We obtained approximately 6 million sequence tags per sample,and 6018254, 5930726, 6022170, 5950123, 5991210, 5798939, 5823113, 5772449,5858527 and 5657697 clean tags were obtained in libraries of B. rapa, B. oleracea, B. napus-F1, B. napus-F2, B. napus-F3, B. napus-F4, natural B. napus, B. nigra, B. juncea and B. carinata, respectively.16574, 15970, 22059, 18155, 16479, 18196, 17448, 13867, 19424 and 16645 genes of B. rapa genome were unambigously mapped by sequence tags of these ten DGE libraries, respectively. Differentially expressed genes during polyploidization were broadly discovered by comparing the tetraploids with their progenitors.

2013-06-27 | GSE43246 | GEO

Gene expression profilling of six species of Brassica and early four generation of resynthesized B. napus (F1-F4)

2013-06-27 | E-GEOD-43246 | biostudies-arrayexpress

transilice - Effects of silicon (Si) supply on leaf and root transcriptomes

Project description:ngs2017_08_brasilice-transilice - What are the genes with modulated expression in response to brassica napus treatment (1.7mM, One week, root supply)-Brassica napus were grown on hydropnic conditions using Hoagland nutrient solution containing or not 1.7 mM of Si

2019-02-22 | GSE122968 | GEO

Identifying conserved and novel miRNAs in developing seeds of Brassica napus using deep sequencing [small RNA-seq]

Project description:MicroRNAs (miRNAs) are important post-transcriptional regulators of plant development and seed formation. In Brassica napus, an important edible oil crop, valuable lipids are synthesized and stored in specific seed tissues during embryogenesis. The miRNA transcriptome of B. napus is currently poorly characterized, especially at different seed developmental stages. This work aims to describe the miRNAome of developing seeds of B. napus by identifying plant-conserved and novel miRNAs and comparing miRNA abundance in mature versus developing seeds. A total of 62 miRNA families were detected through a computational analysis of a large number of reads obtained from deep sequencing two small RNA and two RNA-seq libraries of (i) pooled immature developing stages and (ii) mature B. napus seeds. Among these miRNA families, 17 families are currently known to exist in B. napus; additionally, 32 families not reported in B. napus but conserved in other plant species were identified by alignment with known plant mature miRNAs. The contigs from the assembled mRNA-seq data allowed for a search for putative new precursors and led to the identification of 13 novel miRNA families. Differential expression between the libraries was determined through a statistical analysis of normalized miRNA reads and revealed several miRNAs and isomiRNAs that were more abundant during the developing stages. The predicted miRNA target genes encode a broad range of proteins related to seed development and energy storage. This work presents a comprehensive study of the miRNA transcriptome of B. napus seeds and will provide a basis for future research on more targeted studies of individual miRNAs and their functions in embryogenesis, seed maturation and lipid accumulation in B. napus. microRNA profiles in 2 different seed libraries (mature seeds and a pool of developing seed stages) of Brassica napus by deep sequencing (Illumina HiSeq2000).

2014-08-01 | E-GEOD-38019 | biostudies-arrayexpress

Identifying conserved and novel miRNAs in developing seeds of Brassica napus using deep sequencing [mRNA-seq]

Project description:MicroRNAs (miRNAs) are important post-transcriptional regulators of plant development and seed formation. In Brassica napus, an important edible oil crop, valuable lipids are synthesized and stored in specific seed tissues during embryogenesis. The miRNA transcriptome of B. napus is currently poorly characterized, especially at different seed developmental stages. This work aims to describe the miRNAome of developing seeds of B. napus by identifying plant-conserved and novel miRNAs and comparing miRNA abundance in mature versus developing seeds. A total of 62 miRNA families were detected through a computational analysis of a large number of reads obtained from deep sequencing two small RNA and two RNA-seq libraries of (i) pooled immature developing stages and (ii) mature B. napus seeds. Among these miRNA families, 17 families are currently known to exist in B. napus; additionally, 32 families not reported in B. napus but conserved in other plant species were identified by alignment with known plant mature miRNAs. The contigs from the assembled mRNA-seq data allowed for a search for putative new precursors and led to the identification of 13 novel miRNA families. Differential expression between the libraries was determined through a statistical analysis of normalized miRNA reads and revealed several miRNAs and isomiRNAs that were more abundant during the developing stages. The predicted miRNA target genes encode a broad range of proteins related to seed development and energy storage. This work presents a comprehensive study of the miRNA transcriptome of B. napus seeds and will provide a basis for future research on more targeted studies of individual miRNAs and their functions in embryogenesis, seed maturation and lipid accumulation in B. napus. RNA profiles in 2 different seed libraries (mature seeds and a pool of developing seed stages) of Brassica napus by deep sequencing (Illumina HiSeq2000).

2014-08-01 | E-GEOD-38018 | biostudies-arrayexpress

Transcription profiling of Brassica lines

2010-05-19 | E-GEOD-15915 | biostudies-arrayexpress

Production of Red Flower Rapeseed by Ectopic Expression of OvPAP2 Identified in Brassica napus-Orychophragmus violaceus Addition Line by Comparative Transcriptome Analysis

Project description:Oilseed rape is both an important oleaginous crop and agriculture sightseeing crop whereas has relatively scanty flower color. As natural ﬂavonoids, Anthocyanin are responsible for the attractive red, purple, and blue colors of various tissues in higher plants, especially for the ornamental plants flower. One Brassica napus-Orychophragmus violaceus disomic addition line (M4) obtained previously exhibits red petals whichresult from anthocyanin biosynthesis. Transcriptome analysis of M4, B. napus (H3), natural individuals of O. violaceus with purple petals (OvP) and white petals (OvW) revealed that most of structural genes for the anthocyanin synthesis were up-regulated in both M4 and OvP, especially key gene ANS in the last step. Reads assembling and sequence alignment showed that the regulatory DEG PAP2 in M4 was from the transcript of O. violaceus. OvPAP2 was transformed into Arabidopsis thaliana and B. napus driven by the CaMV35S promoter and the rape petal-specific prompter XY355. Transgenic A. thaliana plants showed different levels of purple pigments in most of the organs, including the petals, and transgenic B. napus flowers exhibited restricted accumulation of anthocyanins in stamens when driven by CaMV35S promoter, but generated both red petals and anthers driven by the XY355 promoter. These results provided a platform for expounding the anthocyanin biosynthesis pathway in B. napus petals and give a successful case for flower color modification of the agriculture sightseeing rape.

2017-12-30 | GSE93089 | GEO

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