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Rice (Oryza sativa) RNA-seq 12 samples DESeq2 HISAT2

RNA-Seq Differential Expression Analysis of Rice Salt-Alkali Stress

A first-party account of the analysis behind the session report: paired-end FASTQ quality control, alignment, gene quantification, two timepoint-specific differential-expression contrasts, figures, and exportable result tables.

Input12 samples
Alignment96–97%
Strict DEGs3,456
Reported runtime5,417 s
01 / Scientific question

How does the rice transcriptome respond across early and later salt-alkali stress?

The analysis compared Na₂CO₃-treated and control Nipponbare rice at Day 1 and Day 5. Each condition contained three biological replicates, allowing the two treatment effects to be estimated separately rather than collapsing time into one contrast.

Source dataset
NCBI BioProject PRJNA895747
Experimental design
4 groups × 3 biological replicates
Input format
Paired-end RNA-seq FASTQ plus sample metadata
Reference
Oryza sativa japonica IRGSP-1.0
02 / Managed execution

Workflow and methods

The report records the toolchain, versions, statistical thresholds, and files passed between steps so the execution path can be inspected.

Read quality controlfastp 0.23.2 detected and removed adapters, trimmed low-quality bases, and discarded reads shorter than 36 nt.
Reference alignmentHISAT2 2.2.1 aligned reads to IRGSP-1.0; SAMtools sorted and indexed the alignment files.
Gene quantificationfeatureCounts 2.0.3 counted properly paired reads overlapping annotated exons.
Differential expressionDESeq2 1.38.0 tested Day 1 and Day 5 treatment-versus-control contrasts independently.
Evidence packageNormalized counts, ranked gene lists, overlap tables, QC summaries, heatmaps, PCA, and volcano plots were exported with the report.
03 / Results

A larger strict response was detected at Day 5

Using the report’s strict threshold of FDR ≤ 0.01 and |log₂ fold change| ≥ 0.585, the analysis identified 1,451 differentially expressed genes at Day 1 and 2,005 at Day 5.

Day 1

649 upregulated and 802 downregulated genes.

Day 5

869 upregulated and 1,136 downregulated genes.

Shared response

331 strict DEGs overlapped between timepoints.

Temporal specificity

1,120 genes were Day-1-specific and 1,674 were Day-5-specific.

Four-panel chart comparing tested genes, strict differential expression, direction of change, and overlap between Day 1 and Day 5 rice salt-alkali stress responses
Report figure: comparison of differential-expression counts, regulation direction, fold-change distributions, and strict DEG overlap across the two timepoints.
04 / Interpretation boundary

Limitations and validation status

  • The page reports computational results from this dataset; it does not claim independent experimental validation of individual genes.
  • The session report exports enrichment-ready gene lists and ranked inputs, but it does not present interpreted GO or KEGG enrichment results. Those should not be implied as completed findings.
  • The reported 331-gene overlap is based on the strict statistical threshold and should not be read as the complete biological response shared between timepoints.
  • Software versions and thresholds reflect this recorded run and should be rechecked before reproducing the analysis in a different environment.
Evidence status

Reproduced by Pipette from public source data. Findings remain computational until supported by independent biological validation.

05 / Provenance

Reproducibility artifacts recorded in the session

The downloadable report lists the inputs, tools, versions, thresholds, output filenames, and generated figures. Representative artifacts include:

count_matrix.csv38,993 genes across 12 samples
de_results_day1/Complete Day 1 DESeq2 result set
de_results_day5/Complete Day 5 DESeq2 result set
overlap_genes_strict.csv331 genes significant at both timepoints
gsea_ranked_day1.csvRanked Day 1 genes for downstream analysis
sample_qc_table.csvRead depth, detection, and alignment metrics
Original session report14-page PDF containing the recorded prompt, results, methods, output inventory, and figures.
Download PDF

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