Summary: Plant growth is severely affected by toxic concentrations of the non-essential heavy metal cadmium (Cd). Comprehensive transcriptome analysis by RNA-Seq following cadmium exposure is required to further understand plant responses to Cd and facilitate future systems-based analyses of the underlying regulatory networks. In this study, rice plants were hydroponically treated with 50 µM Cd for 24 hours and ~60,000 expressed transcripts, including transcripts that could not be characterized by microarray-based approaches, were evaluated. Upregulation of various ROS-scavenging enzymes, chelators and metal transporters demonstrated the appropriate expression profiles to Cd exposure. Gene Ontology enrichment analysis of the responsive transcripts indicated the upregulation of many drought stress-related genes under Cd exposure. Further investigation into the expression of drought stress marker genes such as DREB suggested that expression of genes in several drought stress signal pathways was activated under Cd exposure. Furthermore, qRT-PCR analyses of randomly selected Cd-responsive metal transporter transcripts under various metal ion stresses suggested that the expression of Cd-responsive transcripts might be easily affected by other ions. Our transcriptome analysis demonstrated a new transcriptional network linking Cd and drought stresses in rice. Considering our data and that Cd is a non-essential metal, the network underlying Cd stress responses and tolerance, which plants have developed to adapt to other stresses, could help to acclimate to Cd exposure. Our examination of this transcriptional network provides useful information for further studies of the molecular mechanisms of plant adaptation to Cd exposure and the improvement of tolerance in crop species.
Overall Design: Rice roots and shoots grown in culture medium 0, 1 and 24 hours with 50 µM CdSO4 for Cd stress.
Strategy: |
|
Species: |
|
Tissue: |
|
Development Stage: |
|
Growth Protocol: | Rice (Oryza sativa ssp. japonica cv. Nipponbare) seeds were germinated and grown by hydroponic culture in nutrient media [1.425 mM NH4NO3, 0.323 mM NaH2PO4, 0.513 mM K2SO4, 0.998 mM CaCl2, 1.643 mM MgSO4, 0.009 mM MnCl2, 0.075 mM (NH4)6 Mo7O24, 0.019 mM H3BO3, 0.155 mM CuSO4, 0.036 mM FeCl3, 0.070 mM citric acid, and 0.152 mM ZnSO4] [23] in a growth chamber at 28°C and 70–80% humidity in a 16h light/8h dark cycle. |
Treatment Protocol: | After 10 days, the seedlings of uniform size and growth were subjected to Cd stress treatment by transferring them to a similar medium with 50 µM CdSO4. The plants were maintained under Cd stress conditions for 120 h and then details of plant growth were recorded and sampling was performed as described previously |
Extract Protocol: | Total RNA was extracted from all tissue samples using an RNeasy Plant Kit (Qiagen, Hilden, Germany) according to manufacturer's instructions. |
Library Construction Protocol: | Construction of 13 cDNA libraries (2 tissues, 3 conditions, and 2–3 replicates) from total RNA using a TruSeq RNA sample preparation kit and sequencing with the Illumina Genome Analyzer IIx (Illumina Inc., San Diego, CA, USA) was performed according to the manufacturer's protocols. |
Molecule Type: | poly(A)+ RNA |
Library Source: | |
Library Layout: | SINGLE |
Library Strand: | Reverse |
Platform: | ILLUMINA |
Instrument Model: | Illumina Genome Analyzer IIx |
Strand-Specific: | Specific |
Data Resource | GEN Sample ID | GEN Dataset ID | Project ID | BioProject ID | Sample ID | Sample Name | BioSample ID | Sample Accession | Experiment Accession | Release Date | Submission Date | Update Date | Species | Race | Ethnicity | Age | Age Unit | Gender | Source Name | Tissue | Cell Type | Cell Subtype | Cell Line | Disease | Disease State | Development Stage | Mutation | Phenotype | Case Detail | Control Detail | Growth Protocol | Treatment Protocol | Extract Protocol | Library Construction Protocol | Molecule Type | Library Layout | Strand-Specific | Library Strand | Spike-In | Strategy | Platform | Instrument Model | Cell Number | Reads Number | Gbases | AvgSpotLen1 | AvgSpotLen2 | Uniq Mapping Rate | Multiple Mapping Rate | Coverage Rate |
---|