Gene Expression
Nebulas
Gene Expression NebulasSummary: Flag (FL) and second leaves (SL) in rice show differential aging patterns during monocarpic senescence. Coordination of aging programs in the top leaves is important for effective grain-filling. However, molecular bases for differential aging programs in the top leaves have not been systematically explored in rice. Here, we performed mRNA-sequencing of FL and SL at six time points during the grain-filling period. mRNA expression data revealed 6,365 genes showing aging-dependent expression changes in FL and/or SL. Of them, while 3047 genes showed shared aging-dependent expression patterns between FL and SL, 3058 genes showed differential expression patterns, which were classified into 5 major groups (G1-5) based on their differential expression patterns. Of the groups, G3 representing amino acid (AA) transport showed consistent differential age-dependent expression patterns in independent samples, whereas the other groups showed inconsistent differential expression patterns. Moreover, of AA transporters (AATs) in G3, long-distance AATs showed invariant differential age-dependent expression patterns after panicle removal, consistent to panicle removal-invariant differential nitrogen contents between FL and SL, known to be associated with protein concentration in grains. Our results suggest that long-distance AA transport is an invariant core transcriptional program of differential aging in rice top leaves for nitrogen remobilization during grain-filling.
Overall Design: mRNA expression profiles of the flag leaf and 2nd leaf of Oryza Sativa during grain filling (Day After Heading, DAH, 4-44) were examined by Illumina Hi-seq 2500.
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| Growth Protocol: | - |
| Treatment Protocol: | mRNA expression profiles of the flag leaf and 2nd leaf of Oryza Sativa during grain filling (Day After Heading, DAH, 4-44) were examined by Illumina Hi-seq 2500. |
| Extract Protocol: | Rice leaf tissue was ground using a mortar and pestle, and total RNA was extracted from ~100 mg of tissue using RNeasy Plant Mini Kit (Qiagen). Total RNA integrity was confirmed using the Agilent Technologies 2100 BioAnalyzer (RNA integrity number >8.0). Poly(A) mRNA isolation from total RNA and fragmentation were performed using the Illumina Truseq Stranded mRNA LT Sample Prep Kit with poly-T oligo-attached magnetic beads, according to the manufacturer’s instructions. |
| Library Construction Protocol: | Libraries were prepared for multiplex sequencing using Illumina Truseq Stranded mRNA LT Sample Prep Kit, according to the manufacturer’s protocol. |
| Molecule Type: | poly(A)+ RNA |
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| Library Layout: | PAIRED |
| Library Strand: | Forward |
| Platform: | ILLUMINA |
| Instrument Model: | Illumina HiSeq 2500 |
| Strand-Specific: | Specific |