RNA splicing and spliceosome assembly in eukaryotes occur mainly during transcription.However,co-transcriptional splicing has not yet been explored in plants.Here,we built transcriptomes of nascent chromatin RNAs in Arabidopsis thaliana and showed that nearly all introns undergo co-transcriptional splicing,which occurs with higher efficiency for introns in protein-coding genes than for those in noncoding RNAs.Total intron number and intron position are two predominant features that correlate with co-transcriptional splicing efficiency,and introns with alternative 5'or 3'splice sites are less efficiently spliced.Furthermore,we found that mutations in genes encoding trans-acting proteins lead to more introns with increased splicing defects in nascent RNAs than in mature RNAs,and that introns with increased splicing defects in mature RNAs are inefficiently spliced at the co-transcriptional level.Collectively,our results not only uncovered widespread co-transcriptional splicing in Arabidopsis but also identified features that may affect or be affected by co-transcriptional splicing efficiency.
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