biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsSu-kyoung LeeIllumina NovaSeq 6000RNA-seq of coding RNAOryza sativa Japonica GroupOryza sativa Japonica Grouphttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-15310OJAP_WD199 was annotated as partial WD40 genes, whereas showed strong pollen-specific expression. We generated CRISPR-Cas9-induced loss-of-function mutants for OJAP_WD199 and identified homozygous lines with mutations at two independent target sites per gene. Compared to wild-type (WT) plants, fertility was markedly reduced in knockout mutants, a severe reduction to around 20%. Despite these defects, morphological examination of flowers, anthers, and pollen at the pollen maturation stage revealed no obvious differences between the WT and mutant plants. For the corresponding trascriptomic analysis, we sampled mature pollen anthers from a control group and mutant group.biostudies-arrayexpressGrowth Protocol - Plants were transplanted to paddy field after two weeks in incubator at BOD incubator at end of May. The plants were then grown for 3 months in the field.Library Construction - The sequencing library is prepared by random fragmentation of the DNA or cDNA sample, followed by 5' and 3' adapter ligation. Alternatively, \\"tagmentation\\" combines the fragmentation and ligation reactions into a single step that greatly increases the efficiency of the library preparation process. Adapter-ligated fragments are then PCR amplified and gel purified.Nucleic Acid Extraction - Mature anthers were frozen in LN2 and finely ground using mortar. Grounded samples were homogenized using TRI reagents (RNAiso, Takara) and BCP (1-bromo-3-chloropropane). DNA, RNA, and protein layers were separated by centrifugation. Then, total RNA was extracted using the RNeasy Mini Kit (QIAGEN).Sequencing - We used the Illumina platform to generate sequence reads and sequencing was done in such a way that each of the control and OJAP_WD199 groups includes three biological replicates. On average, six gb Illumia paired-end reads were produced for each sample. In the pipeline, the adapter trimming and read pre-processing was carried out using the trim_galore tool (https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/). After quality assessment of filtered reads with fastqc, reads were mapped to the Rice reference genome (RGAP version 7) using the Hisat2 aligner. The resultant read mapped file was processed using featureCounts and DEGs were estimated from the raw count table using the DESeq2 package.Sample Collection - The samples were collected from paddy field at anthesis stage. We froze the anthers immediately using liquid nitrogen.OrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesSu-kyoung LeeKi-Hong JungfalseRNA sequencing of control and OJAP_WD199 knock out mutant in mature rice anthers. (oryza sativa jap.)OJAP_WD199 was annotated as partial WD40 genes, whereas showed strong pollen-specific expression. We generated CRISPR-Cas9-induced loss-of-function mutants for OJAP_WD199 and identified homozygous lines with mutations at two independent target sites per gene. Compared to wild-type (WT) plants, fertility was markedly reduced in knockout mutants, a severe reduction to around 20%. Despite these defects, morphological examination of flowers, anthers, and pollen at the pollen maturation stage revealed no obvious differences between the WT and mutant plants. For the corresponding trascriptomic analysis, we sampled mature pollen anthers from a control group and mutant group.2026-01-01T00:00:00Z2026-01-02T02:02:31.83Z2025-07-03T16:48:16.983ZE-MTAB-15310ERP174610EFO_0002944EFO_0004170EFO_0003789EFO_0005518EFO_0003738EFO_0004184