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SRX2265965: GSM2358978: T35_3; Vitis vinifera; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 19.7M spots, 5.9G bases, 2.3Gb downloads

Submitted by: NCBI (GEO)
Study: Integration of transcriptomic and proteomic analyses reveals new insights into the response of grape leaves to high temperature
show Abstracthide Abstract
Heat stress is one of the primary abiotic stresses that limit crop production . Grape is a popular cultivated fruit with high economic value throughout the world, and whose growth and development is often influenced by high temperature. Alternative splicing (AS) is a widespread mechanism increasing transcriptome complexity and proteome diversity. We conducted high temperature treatments (35oC, 40oC and 45oC) on grapevines (Vitis vinifera), and assessed proteomic and transcriptomic (especially AS)changes in leaves. We found that nearly 70% of the genes were alternatively spliced under high temperature. Intron retention (IR), exon skipping (ES) and alternative donor/acceptor sites were markedly induced under different high temperatures. IR was the most abundant up- and down-regulated AS event; moreover, IR events at 40 and 45oC were far higher than those at 35oC. These results indicated AS, especially IR, is an important posttranscriptional regulatory during grape leaf responses to high temperature. Proteomic analysis showed that protein levels of the RNA binding proteins SR45, SR30, and SR34, and the nuclear ribonucleic protein U1A in grape leaves gradually rose as ambient temperature increased. The results also revealed why AS events occurred more frequently under high temperature in grape leaves. After integrating transcriptomic and proteomic data, we found that HSPs and some important transcript factors such as MBF1c and HSFA2 were mainly involved in heat tolerance in grape through up-regulating transcriptional and translational levels, and were especially modulated by AS. The results provide the first simultaneous evidence for grape leaf responses to high temperature at transcriptional, posttranscriptional and translational levels. Overall design: We conducted high temperature treatments (35oC, 40oC and 45oC) on grapevines (Vitis vinifera), and assessed proteomic and transcriptomic (especially AS)changes in leaves
Sample: T35_3
SAMN05936449 • SRS1757489 • All experiments • All runs
Organism: Vitis vinifera
Library:
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNAs from leaves were extracted using Trizol (Invitrogen, 15596-018) according to the manufacturer’s instructions. Highly purified and intact mRNAs were enriched from total RNAs using Dynabeads® mRNA purification kit (Ambion, 61006). RNAs were fragmented into ~ 300 nt fragments by 1 min incubation at 94℃ in fragmentation buffer (10 mM ZnCl2, 10 mM Tris-HCl, pH 7.0). The fragmentation reaction was stopped with 50 mM EDTA, followed by standard ethanol precipitation and collected for sequencing and expression validation. For the strand-specific RNA-seq, fragmented RNAs were re-suspended in H2O and used for library generation with mRNA sequencing kit (Illumina Inc. San Diego, CA). Sequencing was carried out on Illumina HiSeq 4000 sequencer (Illumina Inc. San Diego, CA) according to the manufacturer’s instructions and 150 nt paired-end sequencing reads were generated.
Experiment attributes:
GEO Accession: GSM2358978
Links:
Runs: 1 run, 19.7M spots, 5.9G bases, 2.3Gb
Run# of Spots# of BasesSizePublished
SRR444714519,677,8925.9G2.3Gb2017-03-09

ID:
3321535

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