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SRX13951778: GSM5840789: Leaf control replicate 1; Arundo donax; RNA-Seq
1 ILLUMINA (Illumina HiSeq 2000) run: 38.4M spots, 11.5G bases, 3.4Gb downloads

External Id: GSM5840789_r1
Submitted by: Plant genetics, Di3A, University of Catania
Study: Global leaf and root transcriptome reprogramming in response to cadmium reveals tolerance mechanisms in Arundo donax L.
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The expected increase of sustainable energy demand has shifted the attention towards bioenergy crops. Due to their know tolerance against abiotic stress and relatively low supply request, they have been proposed as election crops to be cultivated in marginal lands without disturbing the part of lands employed for agricultural purposes. Arundo donax L. is a promising bioenergy crop whose behaviour under water and salt stress has been recently studied at transcriptomic levels. As the anthropogenic activities produced in the last years a worrying increase of cadmium contamination worldwide, the aim of our work was to decipher the global transcriptomic response of A. donax leaf and root in the perspective of its cultivation in contaminated soil. In our study, RNA-seq libraries yielded a total of 416 million clean reads and 10.4 Gb per sample. De novo assembly of clean reads resulted in 378,521 transcripts and 126,668 unigenes with N50 length of 1812 bp and 1555 bp, respectively. Differential gene expression analysis revealed 5,303 deregulated transcripts (3,206 up- and 2,097 down regulated) specifically observed in the Cd-treated roots compared to Cd-treated leaves. Among them, we identified genes related to “Protein biosynthesis”, “Phytohormone action”, “Nutrient uptake”, “Cell wall organisation”, “Polyamine metabolism”, “Reactive oxygen species metabolism” and “Ion membrane transport”. Globally, our results indicate that ethylene biosynthesis and the downstream signal cascade are strongly induced by cadmium stress. In accordance to ethylene role in the interaction with the ROS generation and scavenging machinery, the transcription of several genes (NADPH oxidase 1, superoxide dismutase, ascorbate peroxidase, different glutathione S-transferases and catalase) devoted to cope the oxidative stress is strongly activated. Several small signal peptides belonging to ROTUNDIFOLIA, CLAVATA3, and C-TERMINALLY ENCODED PEPTIDE 1 (CEP) are also among the up-regulated genes in Cd-treated roots functioning as messenger molecules from root to shoot in order to communicate the stressful status to the upper part of the plants. Finally, the main finding of our work is that genes involved in cell wall remodelling and lignification are decisively up-regulated in giant reed roots this being a mechanism of cadmium avoidance adopted by giant cane and strongly supporting its cultivation in cadmium contaminated soils in a perspective to save agricultural soil for food and feed crops. Overall design: Examination of Arundo donax response to cadmium treatment. The experimental was conducted using G10 ecotype of Arundo donax. Three control plants were transplanted in pots with simple soil and they were irrigated with water containing 5.777g/L of ammonium nitrate three times per week. Soil contamination of three trated plants was obtained by adding 4 ppm (4 mg/kg) of cadmium nitrate to the soil and they were irrigated with tap water trhee times per week. Leaves and roots were sampled from both control and treated plants.
Sample: Leaf control replicate 1
SAMN25338423 • SRS11792211 • All experiments • All runs
Organism: Arundo donax
Library:
Name: GSM5840789
Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: On July 28th both treated and control samples were collected after prolonged cadmium exposure. Leaves and roots were immediately frozen with liquid nitrogen and stored at -80°C until further uses. RNA isolation was carried out by using the Spectrum Plant Total RNA Extraction kit (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's instructions. All samples passed through three steps before library construction: Nanodrop for preliminary quantitation, agarose gel electrophoresis to tests RNA degradation and potential contamination, Agilent 2100 to checks RNA integrity and quantitation. After the QC procedures, mRNA from is enriched from total RNA using oligo(dT) beads. The mRNA is then fragmented randomly in fragmentation buffer, followed by cDNA synthesis using random hexamers and reverse transcriptase. After first-strand synthesis, a custom second-strand synthesis buffer (Illumina) is added, with dNTPs, RNase H and Escherichia coli polymerase I to generate the second strand by nick-translation and AMPure XP beads is used to purify the cDNA. The final cDNA library is ready after a round of purification, terminal repair, Atailing, ligation of sequencing adapters, size selection and PCR enrichment. Libraries are fed into Illumina machines according standard Illumina protocol.
Runs: 1 run, 38.4M spots, 11.5G bases, 3.4Gb
Run# of Spots# of BasesSizePublished
SRR1778958638,370,20111.5G3.4Gb2022-02-03

ID:
19469297

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